{"gene":"CRKL","run_date":"2026-06-09T22:57:19","timeline":{"discoveries":[{"year":1993,"finding":"CRKL was identified as a novel gene on chromosome 22q11 encoding a 303 amino acid protein with one SH2 and two SH3 domains, structurally related to but distinct from c-CRK (v-crk oncogene homolog), predicted to function as a signal transduction adaptor.","method":"cDNA cloning, sequence analysis","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — direct sequencing and domain identification in a single foundational study; no functional validation beyond sequence","pmids":["8361759"],"is_preprint":false},{"year":1994,"finding":"CRKL is the major tyrosine-phosphorylated protein (39 kDa) in CML neutrophils and cell lines expressing p210BCR-ABL, and a direct interaction between CRKL and ABL was demonstrated by yeast two-hybrid screen.","method":"Anti-phosphotyrosine immunoblotting, protein purification, microsequencing, yeast two-hybrid","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — independently replicated by multiple labs (PMID 8083188, 7524758, 7521685) with orthogonal methods","pmids":["8083188","7524758","7521685"],"is_preprint":false},{"year":1994,"finding":"CRKL is a substrate for the p210BCR/ABL and p145ABL kinases in vivo; BCR/ABL and ABL co-immunoprecipitate with CRKL, forming stable complexes; mSOS1 also co-immunoprecipitates with CRKL, establishing a putative signaling pathway from BCR/ABL through CRKL to SOS.","method":"Co-immunoprecipitation, in vitro kinase assay","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP and kinase assay, replicated across multiple subsequent studies","pmids":["8168080"],"is_preprint":false},{"year":1995,"finding":"The CRKL SH2 domain binds specifically to tyrosine-phosphorylated CBL in Ph-positive leukemia cells; CRKL SH3 domains bind BCR/ABL but not CBL; a trimolecular complex of BCR/ABL–CRKL–CBL exists in Ph-positive cells.","method":"GST fusion protein pulldown, co-immunoprecipitation, domain mapping","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP with domain mapping, replicated in multiple cell contexts","pmids":["7545163"],"is_preprint":false},{"year":1995,"finding":"CRKL SH3 domains bind c-ABL and p210BCR/ABL; CRKL SH2 domain binds paxillin (a focal adhesion protein) at tyrosines 31 and 118 (but not 181), physically linking p210BCR/ABL to paxillin in CML cells.","method":"GST-CRKL fusion protein binding assay, site-directed mutagenesis, co-immunoprecipitation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro binding with mutagenesis identifying specific phosphotyrosine residues, single lab","pmids":["7493940"],"is_preprint":false},{"year":1996,"finding":"CRKL SH2 domains bind p120CBL; CRKL and c-CRK SH3 domains bind BCR/ABL and c-ABL; CRKL participates in multimeric complexes including p120CBL, PI3K, and BCR/ABL, linking BCR/ABL to the PI3K pathway.","method":"In vitro binding studies, co-immunoprecipitation, PI3K lipid kinase activity assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vitro domain binding plus functional PI3K activity assay in co-precipitates, single lab with multiple orthogonal methods","pmids":["8632906"],"is_preprint":false},{"year":1996,"finding":"CRKL overexpression activates RAS and JUN kinase signaling pathways and transforms fibroblasts in a RAS-dependent manner. CRKL contributes to BCR-ABL fibroblast transformation alongside GRB2; deletion of both CRKL and GRB2 binding sites reduced transforming activity 15-fold.","method":"Fibroblast transformation assay, RAS activation assay, BCR-ABL deletion mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — functional transformation assay with domain deletion mutants and epistasis, single lab with multiple orthogonal methods","pmids":["8798523"],"is_preprint":false},{"year":1996,"finding":"CRKL SH2 domain binds tyrosine-phosphorylated p130CAS in BCR/ABL-transformed cells; p130CAS is tyrosine-phosphorylated and constitutively associated with CRKL in CML cells; BCR/ABL disrupts the normal interaction between p130CAS and tensin but not p130CAS–FAK or p130CAS–paxillin.","method":"Co-immunoprecipitation, immunofluorescence","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP in cell lines and patient samples, single lab","pmids":["8810278"],"is_preprint":false},{"year":1997,"finding":"Tyrosine 207 in CRKL (between the two SH3 domains) is the major BCR/ABL phosphorylation site in vivo; Y207F mutation eliminates tyrosine phosphorylation; phosphorylation at Y207 provides a binding site for the CRKL SH2 domain itself (autoinhibitory interaction) but does not alter stoichiometry of SOS or C3G complex formation.","method":"In vitro kinase assay, site-directed mutagenesis, tryptic phosphopeptide mapping","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay with mutagenesis identifying specific phosphorylation site, single lab","pmids":["9053848"],"is_preprint":false},{"year":1997,"finding":"CRKL N-terminal SH3 domain directly binds BCR/ABL at a proline-rich C-terminal region of ABL; however, deletion of this proline-rich region did not impair BCR/ABL factor-independent transformation of myeloid cells, and CRKL still became phosphorylated via indirect interaction with the mutant BCR/ABL.","method":"Yeast two-hybrid, gel overlay, co-immunoprecipitation, factor-independent growth assay","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding mapping with negative functional result for direct interaction requirement, single lab","pmids":["8978305"],"is_preprint":false},{"year":1997,"finding":"After beta1 integrin ligation, CRKL SH2 domain binds tyrosine-phosphorylated p120CBL in megakaryocytic MO7e cells and p110HEF1 in lymphoid H9 cells; CRKL is constitutively complexed to C3G, SOS, and c-ABL via its SH3 domains regardless of integrin ligation, enabling cell-type-specific signaling complexes.","method":"Co-immunoprecipitation, domain-specific binding assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP in two distinct cell lines with domain specificity mapping, single lab","pmids":["9162067"],"is_preprint":false},{"year":1997,"finding":"CrkL N-terminal SH3 domain binds C3G, and co-expression of CrkL with C3G in Cos1 cells significantly increased GTP/GDP ratio on Rap1; CrkL enhances C3G-mediated Rap1 activation primarily by membrane recruitment of C3G, requiring both SH2 and SH3 domains (SH2 requirement compensable by farnesylation signal).","method":"Co-expression/overexpression, GTP-binding assay, farnesylation rescue experiment","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — functional GTPase assay with mechanistic dissection using domain mutants and rescue, single lab","pmids":["9268367"],"is_preprint":false},{"year":1997,"finding":"BCR/ABL-induced leukemogenesis causes tyrosine phosphorylation of Hef1/Cas-L and its association with the CRKL SH2 domain; P190BCR/ABL, CRKL, and Hef1 or p120CBL form complexes in leukemic tissues, supporting a model where CRKL mediates BCR/ABL signaling to beta1-integrin pathway components.","method":"Co-immunoprecipitation, GST-domain binding in transgenic mouse leukemic tissue","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP in primary leukemic tissue but single lab, single method","pmids":["9405482"],"is_preprint":false},{"year":1997,"finding":"Type I interferon (IFNα) induces rapid tyrosine phosphorylation of CRKL; this phosphorylation is regulated by the IFN receptor-associated Tyk-2 kinase, as shown by Tyk-2 kinase activity in anti-CrkL immunoprecipitates and IFNα-dependent association of CrkL with Tyk-2.","method":"Co-immunoprecipitation, in vitro kinase assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP with kinase activity assay, single lab","pmids":["9374471"],"is_preprint":false},{"year":1997,"finding":"CBL binds directly to the SH2 domain of BCR-ABL when CBL is tyrosine-phosphorylated; CRKL mediates an indirect complex between CBL and BCR-ABL, since deletion of the BCR-ABL SH2 domain did not abolish CBL–BCR-ABL complex formation.","method":"Co-immunoprecipitation, domain mapping","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP with domain deletion mutants, single lab","pmids":["9195915"],"is_preprint":false},{"year":1997,"finding":"Steel factor (SF) activation of c-Kit induces CRKL tyrosine phosphorylation; CRKL co-precipitates with c-Kit through CRKL SH3 domains (not SH2) as part of a larger complex containing p85PI3K and p120CBL; CRKL binds directly to p85PI3K in vitro via its SH3 domain.","method":"Co-immunoprecipitation, Far Western blotting with GST-SH3 fusion protein","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro direct binding assay plus in vivo co-IP, single lab","pmids":["9092574"],"is_preprint":false},{"year":1998,"finding":"CRKL Y207F mutation abolishes all in vivo tyrosine phosphorylation and paradoxically enhances CRKL function (complex formation with SH2-binding proteins, JNK signaling, fibroblast transformation), indicating Y207 phosphorylation acts as a negative regulatory site; both SH2 and N-terminal SH3 domains are essential for CRKL biological activity in fibroblasts (transformation) and hematopoietic cells (adhesion).","method":"Tryptic phosphopeptide mapping, site-directed mutagenesis, transformation assay, JNK kinase assay, adhesion assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — mutagenesis with multiple functional readouts, single lab","pmids":["9710592"],"is_preprint":false},{"year":1998,"finding":"Highly selective peptides binding exclusively to the N-terminal SH3 domains of CrkL and Crk (but not Grb2 SH3 or other SH3 domains) were developed and shown to disrupt pre-existing Crk complexes with DOCK180, SOS, and C3G in a concentration-dependent manner.","method":"GST-fusion peptide pulldown, in-solution precipitation with biotinylated peptides, SH3 domain binding specificity panel","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain specificity demonstrated with broad panel of SH3 domains and complex disruption assays, single lab","pmids":["9591773"],"is_preprint":false},{"year":1999,"finding":"IFNα-phosphorylated STAT5 acts as a docking site for the CRKL SH2 domain; CRKL and STAT5 form a complex that translocates to the nucleus and binds the TTCTAGGAA palindromic element in promoters of a subset of IFN-stimulated genes, establishing CRKL as a nuclear adapter regulating gene transcription.","method":"Co-immunoprecipitation, EMSA (electrophoretic mobility shift assay), nuclear fractionation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — co-IP plus functional EMSA with DNA binding, nuclear translocation confirmed, single lab","pmids":["9872990"],"is_preprint":false},{"year":1999,"finding":"HPK1 (hematopoietic progenitor kinase 1) interacts with CRKL in vitro and in vivo via CRKL SH3 domain binding to proline-rich motifs in HPK1; CRKL synergizes with HPK1 to activate JNK; HPK1 phosphorylates CRKL mainly on serine/threonine residues in vitro.","method":"Co-immunoprecipitation, in vitro kinase assay, JNK activation assay, dominant-negative epistasis","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro kinase assay plus in vivo co-IP with functional JNK readout, single lab","pmids":["9891069"],"is_preprint":false},{"year":1999,"finding":"CRKL overexpression enhances spontaneous cell migration requiring both SH2 and N-terminal SH3 domains; after integrin cross-linking, full-length but not ΔSH2 CRKL co-precipitates tyrosine-phosphorylated CBL; the major CRKL SH3-binding protein in hematopoietic Ba/F3 cells is C3G; overexpression of C3G also enhances migration, suggesting a CBL–CRKL–C3G complex in migration signaling.","method":"Transwell migration assay, co-immunoprecipitation with domain deletion mutants, FACS-sorted GFP-tagged constructs","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional migration assay with domain mutants plus protein interaction mapping, single lab","pmids":["10608804"],"is_preprint":false},{"year":1999,"finding":"CrkL overexpression enhances hematopoietic cell adhesion to fibronectin via VLA-4 and VLA-5 integrins without changing integrin surface expression; the N-terminal SH3 domain (C3G binding) is critical, while C-terminal SH3 and Y207 are dispensable; C3G overexpression also increases adhesion and C3G mutant lacking the GEF domain blocks CrkL-induced adhesion, establishing a CrkL–C3G pathway activating integrins.","method":"Adhesion assay, domain-deletion mutant analysis, dominant negative C3G","journal":"Blood","confidence":"High","confidence_rationale":"Tier 2 / Moderate — functional assay with systematic domain deletion and epistasis using dominant-negative C3G, single lab","pmids":["10339478"],"is_preprint":false},{"year":1999,"finding":"CrkL overexpression enhances Epo/IL-3-induced ERK1/2 activation (augmented and prolonged) via a Ras-dependent mechanism through C3G; CrkL mutants lacking SH2 or N-terminal SH3 domains inhibit Epo-induced ERK2 activation; CrkL also modestly activates JNK.","method":"Elk-1 reporter assay, ERK kinase assay, Ras-GTP pulldown, dominant-negative Ras","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional signaling assay with domain mutants and epistasis, single lab","pmids":["10514505"],"is_preprint":false},{"year":2000,"finding":"HGF stimulation induces association of CRKL (via SH2 domain) with phosphorylated YXXP motifs in the docking protein GAB1; CRKL's N-terminal SH3 domain then binds C3G, activating Rap1; CRKL–C3G interaction is required for HGF-induced Rap1 activation and contributes to reduced cell adhesion/migration.","method":"Co-immunoprecipitation, dominant-negative C3G transfection, Rap1-GTP assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain mapping plus functional Rap1 GTPase assay and rescue experiments, single lab","pmids":["10753869"],"is_preprint":false},{"year":2000,"finding":"In BCR-ABL-expressing cells, CRKL interacts with tyrosine-phosphorylated STAT5 and is found in the nucleus, detectable in a STAT5/DNA complex; CRKL increases transcriptional activation from a STAT-responsive reporter, functioning as a nuclear adaptor that associates with and activates STAT proteins.","method":"Co-immunoprecipitation, indirect immunofluorescence, EMSA supershift, luciferase reporter assay","journal":"Experimental hematology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus functional transcriptional assay with nuclear localization, single lab","pmids":["10720695"],"is_preprint":false},{"year":2000,"finding":"SHIP1 forms a signaling complex with DOK1, PI3K, and CRKL in BCR/ABL-transformed cells; DOK1 binds SHIP1 directly via its PTB domain; CRKL directly binds SHIP1 via the CRKL SH2 domain; SHIP1 tyrosines 917 and 1020 mediate interactions with DOK1; expression of tyrosine-mutant SHIP1 fails to alter migration.","method":"Co-immunoprecipitation, direct binding assays, migration assay with tyrosine mutants","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct binding mapped to specific tyrosine residues with functional migration readout, single lab","pmids":["11031258"],"is_preprint":false},{"year":2001,"finding":"Crkl-null mice exhibit defects in cranial/cardiac neural crest derivatives (ganglia, aortic arch arteries, cardiac outflow tract, thymus, parathyroid, craniofacial structures) phenocopying DiGeorge syndrome; neural crest migration and early expansion are unaffected, indicating an essential role for Crkl in neural crest function, differentiation, and/or survival.","method":"Targeted gene knockout (null mutation), embryo phenotypic analysis","journal":"Nature genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean knockout with defined tissue-specific phenotype, multiple organ systems analyzed","pmids":["11242111"],"is_preprint":false},{"year":2001,"finding":"CrkL C-terminal SH3 domain binds CD34 at a membrane-proximal region of the CD34 intracellular tail; this interaction is specific to CrkL (not CrkII); CrkL co-immunoprecipitates with CD34 in vivo.","method":"GST-fusion protein pulldown, co-immunoprecipitation, domain mapping with deletion constructs","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — GST pulldown plus in vivo co-IP confirming C-terminal SH3-mediated interaction, single lab","pmids":["11389015"],"is_preprint":false},{"year":2001,"finding":"CrkL binds WASP through its SH3 domain (binding unaffected by WASP tyrosine phosphorylation) and also associates with Syk tyrosine kinase via both SH2 and SH3 domains; CrkL immunoprecipitates contain kinase-active Syk; CrkL acts as a molecular adapter linking WASP and Syk.","method":"Co-immunoprecipitation, GST fusion protein pulldown, in vitro kinase assay","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain dissection and functional kinase assay, single lab","pmids":["11313252"],"is_preprint":false},{"year":2001,"finding":"CRKL overexpression in BCR/ABL transgenic mice enhances constitutive C3G complex formation, increases integrin-based macrophage motility, and markedly accelerates BCR/ABL-induced leukemia/lymphoma onset; Rap1 is activated in CRKL-overexpressing metastatic tumors.","method":"Transgenic mouse model, co-immunoprecipitation, Rap1 activation assay, migration assay","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo transgenic model with mechanistic co-IP and GTPase assays, single lab","pmids":["11245441"],"is_preprint":false},{"year":2002,"finding":"DOCK2 binds to the N-terminal SH3 domain of CrkL via two separate regions; CrkL and DOCK2 co-localize with F-actin; CrkL-induced Rac1 activation is inhibited by a DOCK2 mutant; DOCK2 associates with Vav in Jurkat cells, revealing a CrkL–DOCK2–Vav complex regulating Rac1.","method":"Co-immunoprecipitation (in vitro and in vivo), immunocytochemistry, Rac1-GTP assay, cell attachment assay","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo binding with functional Rac1 GTPase assay, single lab","pmids":["12393632"],"is_preprint":false},{"year":2002,"finding":"Grit (a RhoGAP for Rho/Rac/Cdc42) directly interacts with TrkA and is also recruited to activated TrkA via binding to N-Shc and CrkL/Crk adapters; overexpression of the TrkA-binding region of Grit inhibited NGF-induced neurite elongation.","method":"Yeast two-hybrid, co-immunoprecipitation, overexpression inhibition assay","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP identifying complex, functional inhibition assay, single lab","pmids":["12446789"],"is_preprint":false},{"year":2002,"finding":"CrkL is required for type I IFN-α-dependent gene transcription via GAS elements (through CrkL:Stat5 complexes); IFN-α-induced Rap1 activation is defective in CrkL-null MEFs; this Rap1 activation requires the N-terminal SH3 domain of CrkL and both Tyk-2 and Jak-1 kinases.","method":"CrkL-/- mouse embryonic fibroblasts, reporter gene assay, Rap1-GTP assay, dominant-negative kinase constructs","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — loss-of-function in null cells with multiple functional readouts and domain rescue, single lab","pmids":["11866427"],"is_preprint":false},{"year":2003,"finding":"Src triggers translocation of CrkL to focal adhesions in a manner dependent on p130Cas; forced localization of CRKL to focal adhesions activates Rac1 and Cdc42 and rescues haptotaxis defects of Src/Yes/Fyn triple-knockout MEFs; CrkL co-recruits Dock1 to focal adhesions for Rac1 activation; MEFs lacking CrkL show impaired integrin-induced migration despite expressing CrkII.","method":"MEF knockout/rescue, live-cell imaging, Rac1/Cdc42 GTPase assay, haptotaxis assay, forced-localization chimeric construct","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic loss-of-function with rescue, GTPase assay, and mechanistic forced-localization experiment, single lab","pmids":["12665586"],"is_preprint":false},{"year":2003,"finding":"CrkL directs ASAP1 (an Arf-GAP) to peripheral focal adhesions; CrkL N-terminal SH3 domain binds ASAP1 in platelets (identified by pulldown/mass spectrometry); co-expression of wild-type CrkL (but not SH2-mutant CrkL, which cannot localize to FAs) recruits ASAP1 to focal adhesions.","method":"Pull-down assay followed by mass spectrometry, co-expression in COS7 cells, immunofluorescence","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pulldown/MS identification with functional localization experiment using SH2 mutant, single lab","pmids":["12522101"],"is_preprint":false},{"year":2004,"finding":"Reelin stimulation causes CrkL (and CrkI/II) to bind tyrosine-phosphorylated Dab1 at Y220 and Y232 (critical for cortical neuron positioning); CrkL also binds C3G; Reelin stimulates tyrosine phosphorylation of C3G and activates Rap1, establishing a Dab1–CrkL–C3G–Rap1 pathway downstream of Reelin.","method":"Affinity pulldown from brain extract, Reelin stimulation of cortical neurons, Rap1-GTP assay, site-specific phosphomutant analysis","journal":"Current biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — biochemical binding mapped to specific pY sites, functional GTPase assay, primary neurons, site-specific mutants required for neuron positioning in vivo","pmids":["15062102"],"is_preprint":false},{"year":2006,"finding":"Compound heterozygosity of Crkl and Tbx1 (mouse homologs of 22q11 genes) drastically increases penetrance of DiGeorge syndrome-like phenotype; Crkl and Tbx1 have dose-dependent functions in pharyngeal segmentation and RA signaling; partial rescue is achieved by genetically reducing RA levels, indicating that del22q11 involves dose-sensitive interaction of CRKL and TBX1 via locally aberrant RA signaling.","method":"Compound heterozygous mouse genetics, epistasis with RA synthesis gene, embryo phenotype analysis","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis in vivo with partial rescue, establishing pathway position and interaction","pmids":["16399080"],"is_preprint":false},{"year":2006,"finding":"Crkl is required for normal cellular responses to Fgf8 (survival, migration, Erk activation, target gene expression); Fgf8 induces tyrosine phosphorylation of FgfR1 and FgfR2 and their direct binding to Crkl; genetic interactions exist between Crkl and Fgf8 in pharyngeal/cardiac morphogenesis.","method":"Crkl-/- mouse embryos, Fgf8 stimulation assay, receptor phosphorylation, Erk activation assay, genetic compound heterozygosity","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function in null cells/embryos with receptor binding and signaling readouts, epistasis with Fgf8","pmids":["16399079"],"is_preprint":false},{"year":2007,"finding":"NS1 proteins of 1918 pandemic and avian H7N3 influenza viruses contain a consensus SH3-binding motif that binds specifically to the N-terminal SH3 domain of CrkL (and Crk) but not to other SH3 domains tested; endogenous CrkL co-precipitates NS1 from H7N3-infected cells; SH3 binding was associated with enhanced PI3K/Akt signaling.","method":"SH3 phage library screening, recombinant protein binding assay, co-immunoprecipitation from infected cells, Akt phosphorylation assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phage library + recombinant binding + co-IP from infected cells with functional signaling readout, single lab","pmids":["18165234"],"is_preprint":false},{"year":2008,"finding":"Reducing expression of Crk and CrkL (but not BDNF pathway) blocks Reelin-stimulated dendritogenesis in hippocampal neurons (approximately twofold enhancement by Reelin); CrkL acts downstream of Dab1 phosphorylation in postnatal hippocampal dendrite development.","method":"Retroviral shRNA knockdown, hippocampal neuron culture, Reelin stimulation, dendrite morphometry","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA knockdown with specific Reelin pathway readout vs. BDNF control, single lab","pmids":["18477607"],"is_preprint":false},{"year":2008,"finding":"The WAVE2 complex recruits Abl kinase upon TCR ligation; WAVE2 regulates TCR-mediated Rap1 activation via membrane recruitment and activation of the CrkL–C3G exchange complex; Abl phosphorylates C3G (required for its GEF activity toward Rap1) but does not regulate CrkL–C3G membrane recruitment; this signaling node regulates integrin clustering and affinity maturation.","method":"Co-immunoprecipitation, Rap1-GTP assay, dominant-negative constructs, membrane fractionation, integrin activation assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods dissecting pathway with specific functional readouts in T cells, single lab","pmids":["18809728"],"is_preprint":false},{"year":2008,"finding":"MUC1 ligation of ICAM-1 induces rapid formation of a Src–CrkL signaling complex at the MUC1 cytoplasmic domain; Src recruits CrkL to MUC1 and is required for CrkL-dependent Rac1- and Cdc42-mediated actin cytoskeletal protrusions at cell–cell contact sites.","method":"Co-immunoprecipitation, Src kinase inhibition, Rac1/Cdc42 GTPase assay, actin reorganization assay","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP with functional actin/GTPase assay, single lab","pmids":["18403635"],"is_preprint":false},{"year":2009,"finding":"CRKL amplification and overexpression in NSCLC activates SOS1-RAS-RAF-ERK and SRC-C3G-RAP1 pathways; CRKL overexpression in EGFR-mutant cells induces resistance to gefitinib by activating ERK and AKT signaling; CRKL knockdown in CRKL-amplified NSCLC induces cell death.","method":"shRNA knockdown, overexpression in airway epithelial cells, anchorage-independent growth assay, pathway inhibitor studies","journal":"Cancer discovery","confidence":"High","confidence_rationale":"Tier 2 / Strong — gain- and loss-of-function with pathway analysis, replicated in multiple NSCLC cell lines","pmids":["22586683"],"is_preprint":false},{"year":2009,"finding":"PI3K p85 subunit interacts with CrkL; this p85–CrkL interaction is required for NKG2D-mediated NK cell cytotoxicity; CrkL depletion impairs NK–target conjugate formation, MTOC polarization, and granule release; Rap1 is activated downstream of NKG2D in a PI3K- and CrkL-dependent manner.","method":"PI3K inhibitor treatment, siRNA depletion of CrkL, conjugate assay, MTOC polarization assay, Rap1-GTP assay","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockdown with multiple functional readouts and signaling pathway dissection, single lab","pmids":["19454690"],"is_preprint":false},{"year":2009,"finding":"Molecular dynamics modeling and in vitro binding studies show that pY463 in FGFR1 binds specifically to the CRKL SH2 domain with ~30-fold higher affinity than to CRK SH2; CRKL (but not CRK) is an essential component of an FGF8-induced feed-forward loop enabling efficient Erk1/2 activation and anchorage-independent growth.","method":"Molecular dynamics simulation, in vitro binding assay with phosphopeptide, Crkl-null cells, Erk activation assay, anchorage-independent growth assay","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — structural modeling validated by in vitro binding affinity measurement, loss-of-function in null cells with functional readouts, single lab","pmids":["19307307"],"is_preprint":false},{"year":2010,"finding":"Agrin stimulates phosphorylation of two tyrosine residues in the C-terminal domain of Dok-7, leading to recruitment of CrkL and Crk; selective inactivation of Crk and CrkL in skeletal muscle causes severe neuromuscular synapse defects in vivo, placing CrkL downstream of MuSK–Dok-7 in presynaptic and postsynaptic differentiation.","method":"In vivo muscle-specific knockout, tyrosine phosphorylation mapping of Dok-7, co-IP","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — in vivo tissue-specific double knockout with clear NMJ phenotype, mapped to specific phosphotyrosine sites on Dok-7","pmids":["21041412"],"is_preprint":false},{"year":2010,"finding":"CRKL is specifically required for p210BCR-ABL-induced IL-3-independent growth of myeloid progenitors and B-lymphoid cell outgrowth; a synthetic phosphotyrosyl peptide binding the CRKL SH2 domain blocks CRKL association with the BCR-ABL complex and reduces c-MYC levels in leukemic cells.","method":"CrkL-null hematopoietic progenitor cells, fetal liver transplantation, SH2-blocking peptide, immunoprecipitation","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — loss-of-function in null progenitor cells plus pharmacological SH2-domain blocking with defined molecular readout, single lab","pmids":["20807813"],"is_preprint":false},{"year":2012,"finding":"NMR and structural analysis reveals CRKL and phospho-CRKL have markedly different structures from CrkII and phospho-CrkII; the SH2 and SH3 domain binding activities are regulated differently in the two proteins; CRKL forms a constitutive complex with Abl (unlike CrkII), explaining the strong preference of Bcr-Abl for CRKL over CrkII.","method":"NMR structural analysis, binding activity assays","journal":"Nature chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structural determination with functional binding validation, explains mechanistic preference for CRKL, single lab","pmids":["22581121"],"is_preprint":false},{"year":2013,"finding":"In rhabdomyosarcoma, CRKL signaling is associated with YES kinase (a Src family kinase); CRKL is required for RMS tumor growth in vitro and in vivo; no interaction of CRKL with IGFIR, MET, or PI3K/AKT/mTOR pathways was detected in this context.","method":"shRNA loss-of-function screen, co-immunoprecipitation, in vivo xenograft assay, kinase pathway inhibitors","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — unbiased shRNA screen validated with in vivo assay and signaling pathway dissection, single lab","pmids":["23318429"],"is_preprint":false},{"year":2013,"finding":"Loss of both Crk and CrkL in fibroblasts (conditional double knockout) causes rounded morphology, loss of focal adhesions, reduced actin stress fibers, microtubule collapse, decreased motility and wound healing, reduced p130Cas phosphorylation; reintroduction of CrkII partially rescues the phenotype; individual deletion of Crk or CrkL alone produces modest phenotype, indicating overlapping but essential roles.","method":"Conditional Crk/CrkL double knockout fibroblasts, morphology and motility assays, immunofluorescence, actin stabilization rescue","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — systematic conditional double knockout with multiple orthogonal readouts and genetic rescue, single lab","pmids":["24166500"],"is_preprint":false},{"year":2014,"finding":"CrkL overexpression enhances lung cancer cell invasion through ERK-MMP9 pathway; CRKL activates AP-1 (c-fos binding to MMP9 promoter) via ERK phosphorylation; ERK inhibitor PD98059 blocks CRKL-induced invasion and MMP9 expression.","method":"Matrigel invasion assay, chromatin immunoprecipitation, luciferase reporter, ERK inhibitor treatment, Western blotting","journal":"Molecular carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional invasion assay with pathway inhibition and promoter occupancy, single lab","pmids":["24664993"],"is_preprint":false},{"year":2014,"finding":"CrkL and CrkL form hetero-oligomers with Crk2; both are recruited to tyrosine-phosphorylated nephrin in podocytes; simultaneous deletion of Crk1/2 and CrkL in podocytes causes albuminuria and altered foot process architecture; CrkL is required for nephrin-induced lamellipodia formation in podocytes in vitro.","method":"Podocyte-specific double conditional knockout mice, co-immunoprecipitation, in vitro lamellipodia assay","journal":"Kidney international","confidence":"High","confidence_rationale":"Tier 2 / Moderate — tissue-specific double knockout with molecular rescue and in vitro mechanistic assay, single lab","pmids":["24499776"],"is_preprint":false},{"year":2015,"finding":"CrkL is required for SDF-1-induced activation of the Raf-1/MEK/Erk pathway through Ras and Rac in hematopoietic cells; CrkL overexpression enhances SDF-1-induced chemotaxis; Lyn mediates SDF-1-induced CrkL tyrosine phosphorylation; the N-terminal SH3 domain (GEF-binding) is required for CrkL-enhanced Erk activation and chemotaxis.","method":"Overexpression, CrkL N-SH3 deletion mutant, Ras/Rac GTP assay, MEK inhibitor, dominant-negative GTPases, Src inhibitor PP1","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple pathway inhibitors with domain deletion mutants and functional chemotaxis assay, single lab","pmids":["16781119"],"is_preprint":false},{"year":2015,"finding":"CRKL is dosage-sensitive for cardiac outflow tract development; haploinsufficiency of CRKL can cause conotruncal heart defects in humans with atypical distal 22q11.2 deletions that exclude TBX1; a Crkl hypomorphic allele series in mice shows spectrum of heart defects dependent on Crkl expression level.","method":"Crkl allelic series (null + hypomorphic), mouse cardiac phenotyping, human genomic deletion mapping","journal":"American journal of human genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — allelic series with dose-dependent phenotype in mice correlated with human genetic evidence","pmids":["25658046"],"is_preprint":false},{"year":2015,"finding":"Sorbs1 and Sorbs2 associate with CrkL (identified by mass spectrometry of CrkL-binding proteins); Sorbs1/2 are required for AChR clustering in vitro and are localized at neuromuscular synapses in vivo, acting as redundant proteins downstream of the MuSK/Dok-7/Crk/CrkL complex.","method":"Mass spectrometry pulldown, CrkL co-immunoprecipitation, AChR clustering assay, in vivo NMJ immunostaining","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — MS-based interaction partner identification with functional validation in vitro and in vivo, single lab","pmids":["26527617"],"is_preprint":false},{"year":2016,"finding":"CRKL is a direct transcriptional target de-repressed by Zeb1-dependent EMT; CRKL serves as a downstream effector of miR-200 loss that mediates focal adhesion formation and outside-in signaling through integrin β1 (ECM→FAK/Src→CRKL) as well as inside-out signaling maintaining tumor cell–matrix contacts; CRKL knockdown suppresses experimental metastases in vivo without affecting primary tumor growth.","method":"miR-200 gain/loss-of-function, CRKL siRNA knockdown, 3D invasion assay, in vivo experimental metastasis model, focal adhesion immunofluorescence","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional loss-of-function with in vivo metastasis readout and mechanistic signaling assays, single lab","pmids":["26728244"],"is_preprint":false},{"year":2017,"finding":"CRKL interacts with tyrosine-phosphorylated p130Cas in PTEN-null cancer cells and is required for p110β-dependent PI3K signaling and cell proliferation in these cells; Src phosphorylates and activates p130Cas, linking PTEN loss to p110β activation via CRKL; Src and p110β inhibition cooperate to suppress PTEN-null cell growth.","method":"CRKL knockdown, co-immunoprecipitation with p130Cas, PI3K signaling assay, Src inhibitor epistasis","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — knockdown with defined signaling pathway readout and epistasis, single lab","pmids":["28723560"],"is_preprint":false},{"year":2017,"finding":"CRKL expression is dosage-sensitive for genitourinary development; Crkl null mice have upper GU defects (23% rate); Crkl heterozygous males have cryptorchidism, lower testis weight, reduced sperm count and subfertility; RNA-seq of Crkl mutant kidneys reveals 52 differentially regulated genes including a 5-fold upregulation of Foxd1 (a nephron progenitor regulator).","method":"Crkl null/heterozygous mice, microdissection phenotyping, RNA-sequencing of developing kidneys","journal":"Proceedings of the National Academy of Sciences","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with transcriptomic pathway analysis in relevant tissue, multiple GU phenotypes documented","pmids":["28439006"],"is_preprint":false},{"year":2018,"finding":"SASH1 interacts with CRKL (identified by yeast two-hybrid and co-IP/MS); SASH1 inhibits CRKL-mediated SRC kinase activation, which is required for EMT; SASH1-deficient cells form significantly more metastases in vivo in a manner entirely dependent on CRKL; CRKL overexpression is associated with poor chemotherapy response.","method":"Yeast two-hybrid, co-immunoprecipitation/mass spectrometry, domain mapping, site-directed mutagenesis, CRISPR knockout, in vivo orthotopic mouse model","journal":"Cellular and molecular gastroenterology and hepatology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods for interaction, in vivo genetic epistasis, CRISPR KO, single lab","pmids":["30480076"],"is_preprint":false},{"year":2024,"finding":"CRKL inhibits APC-mediated proteasomal degradation of β-catenin by competitively decreasing Axin1 binding to APC, thereby promoting VEGFα and CXCL1 expression; this leads to tumor-associated neutrophil infiltration that blocks CD8+ T cell function and creates anti-PD-1 resistance in hepatocellular carcinoma.","method":"Pooled genetic screen, CRISPR-Cas9 knockout, transgenic mouse model (Alb-Cre/Trp53fl/fl + CRKL vector), mass cytometry, co-immunoprecipitation, patient-derived organotypic tumor spheroid model","journal":"Journal of hepatology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal in vitro and in vivo methods with mechanistic co-IP identifying β-catenin/Axin1 interaction, validated in patient models","pmids":["38403027"],"is_preprint":false}],"current_model":"CRKL is an SH2-SH3-SH3 adaptor protein that functions as a major convergence point in tyrosine kinase signaling: its SH2 domain binds phosphotyrosine-containing proteins (pCBL, pCAS/HEF1, paxillin, p130Cas, pGAB1, pDab1, pDok-7, pFGFR1-pY463, pSTAT5, pEpoR), while its N-terminal SH3 domain constitutively binds guanine nucleotide exchange factors (C3G, SOS) and kinases (c-ABL, BCR/ABL), thereby coupling upstream receptor and oncogenic tyrosine kinase signals to Ras/ERK, Rap1, Rac1/Cdc42, and JNK pathways; BCR/ABL phosphorylates CRKL specifically at Y207 (which acts as a negative regulatory autoinhibitory site); in its unphosphorylated state CRKL forms a constitutive complex with Abl (explaining Bcr-Abl's preference for CRKL over CrkII as shown structurally by NMR); CRKL also functions as a nuclear adaptor that complexes with phospho-STAT5 to regulate transcription at GAS elements downstream of type I interferons and cytokines; in developmental contexts CRKL mediates FGF8 receptor signaling (via direct pFGFR1-Y463 binding) and is required downstream of Reelin/Dab1, Dok-7/MuSK, and Src/Cas for neural crest development, neuromuscular synapse formation, podocyte morphogenesis, genitourinary development, and integrin-induced cell migration; additionally, CRKL inhibits APC-mediated β-catenin degradation by displacing Axin1 to activate a VEGFα/CXCL1/tumor-associated neutrophil axis relevant to immune checkpoint resistance."},"narrative":{"mechanistic_narrative":"CRKL is an SH2–SH3–SH3 adaptor protein that couples tyrosine kinase signals to small-GTPase and Ras/ERK pathways, serving as a convergence point in oncogenic, cytokine, and developmental signaling [PMID:8361759, PMID:9710592]. Its modular architecture partitions binding labor: the SH2 domain docks onto phosphotyrosine-containing partners while the N-terminal SH3 domain constitutively engages downstream effectors. Through its SH2 domain CRKL binds tyrosine-phosphorylated CBL, paxillin, p130Cas/HEF1, GAB1, Dab1, Dok-7, the FGFR1 pY463 site, and STAT5 [PMID:7545163, PMID:7493940, PMID:8810278, PMID:10753869, PMID:15062102, PMID:19307307, PMID:21041412], while its N-terminal SH3 domain constitutively binds the guanine-nucleotide exchange factors C3G and SOS and the kinases c-ABL/BCR-ABL [PMID:8168080, PMID:9162067, PMID:9268367]. This dual engagement allows CRKL to relay receptor and oncogene input to Ras/ERK, Rap1, and Rac1/Cdc42 outputs, principally by membrane-recruiting C3G to activate Rap1 and integrins [PMID:9268367, PMID:10339478, PMID:10514505] and by recruiting DOCK family GEFs at focal adhesions to activate Rac1/Cdc42 during integrin- and Src/p130Cas-driven migration [PMID:12393632, PMID:12665586]. In BCR-ABL leukemia CRKL is the major phosphorylated adaptor, forming a constitutive complex with Abl that explains Bcr-Abl's preference for CRKL over CrkII as resolved by NMR; phosphorylation at Y207 acts as a negative-regulatory autoinhibitory site, and CRKL is required for BCR-ABL-driven factor-independent growth [PMID:8083188, PMID:7524758, PMID:7521685, PMID:9053848, PMID:9710592, PMID:20807813, PMID:22581121]. Beyond the cytoplasm, IFNα-activated Tyk-2/Jak-1 phosphorylate STAT5, which docks the CRKL SH2 domain; the CRKL–STAT5 complex translocates to the nucleus and binds GAS-element promoters, making CRKL a nuclear transcriptional adaptor [PMID:9374471, PMID:9872990, PMID:11866427]. CRKL is dosage-sensitive in development: Crkl loss phenocopies DiGeorge syndrome, genetically interacts with Tbx1 through retinoic-acid signaling, and mediates FGF8/FGFR, Reelin/Dab1, and MuSK/Dok-7 signaling required for neural crest, cardiac outflow tract, neuromuscular synapse, podocyte, and genitourinary development [PMID:11242111, PMID:15062102, PMID:16399080, PMID:16399079, PMID:21041412, PMID:24499776, PMID:25658046, PMID:28439006]. In cancer, CRKL amplification activates SOS1-Ras-ERK and Src-C3G-Rap1 to drive NSCLC growth and EGFR-inhibitor resistance, promotes invasion and metastasis, and confers anti-PD-1 resistance in hepatocellular carcinoma by stabilizing β-catenin through displacement of Axin1 from APC [PMID:22586683, PMID:26728244, PMID:30480076, PMID:38403027].","teleology":[{"year":1993,"claim":"Established CRKL as a distinct adaptor-encoding gene, defining its SH2-SH3-SH3 architecture and predicting a signal-transduction role before any function was known.","evidence":"cDNA cloning and sequence/domain analysis of a 22q11 gene","pmids":["8361759"],"confidence":"Medium","gaps":["No functional validation beyond sequence","No binding partners or pathway defined"]},{"year":1994,"claim":"Identified CRKL as the principal tyrosine-phosphorylated adaptor in BCR-ABL leukemia and a direct ABL interactor, anchoring CRKL within oncogenic kinase signaling.","evidence":"Anti-phosphotyrosine immunoblotting of CML cells, yeast two-hybrid, co-IP, and in vitro kinase assays","pmids":["8083188","7524758","7521685","8168080"],"confidence":"High","gaps":["Phosphorylation site on CRKL not yet mapped","Functional consequence of complex formation undefined"]},{"year":1996,"claim":"Mapped CRKL's bipartite binding logic—SH2 to phospho-CBL/paxillin/p130Cas, SH3 to ABL/SOS—and showed CRKL transforms fibroblasts and activates Ras/JNK, defining it as a functional signaling node not a passive scaffold.","evidence":"GST-domain pulldowns, site-directed mutagenesis, co-IP, fibroblast transformation and Ras/JNK activation assays","pmids":["7545163","7493940","8632906","8798523","8810278"],"confidence":"High","gaps":["Endogenous (non-overexpression) requirement not yet tested","Domain contributions to transformation not fully dissected"]},{"year":1997,"claim":"Defined Y207 as the major BCR/ABL phosphorylation site acting as an autoinhibitory negative regulator, and showed the N-SH3–C3G axis drives Rap1 activation via membrane recruitment, establishing the core regulatory and effector mechanisms.","evidence":"Phosphopeptide mapping, Y207F mutagenesis, C3G co-expression with Rap1 GTP-binding and farnesylation-rescue assays","pmids":["9053848","9268367","9162067","9092574"],"confidence":"High","gaps":["Structural basis of Y207 autoinhibition not resolved","Physiological triggers regulating Y207 phosphorylation unclear"]},{"year":1998,"claim":"Confirmed Y207 phosphorylation suppresses CRKL function while SH2 and N-SH3 domains are essential for activity, and identified a CrkL/Crk-selective SH3-binding peptide, providing tools and mechanistic clarity on domain requirements.","evidence":"Y207F mutagenesis with transformation/JNK/adhesion readouts and selective SH3-binding peptide complex-disruption assays","pmids":["9710592","9591773"],"confidence":"High","gaps":["Endogenous regulation of phospho-Y207 in normal cells unaddressed","Peptide selectivity in vivo not established"]},{"year":1999,"claim":"Revealed CRKL as a dual cytoplasmic-nuclear adaptor—coupling Epo/IL-3/integrin signals to Ras/ERK, Rap1, and adhesion via C3G, while also forming nuclear STAT5–DNA complexes at GAS-like elements to regulate IFN-stimulated transcription.","evidence":"Migration/adhesion assays with domain mutants, dominant-negative C3G, ERK/Ras assays, EMSA, nuclear fractionation, HPK1-JNK assays","pmids":["10608804","10339478","10514505","9872990","9891069"],"confidence":"High","gaps":["Mechanism of CRKL nuclear import unknown","Direct vs indirect DNA contact by CRKL not resolved"]},{"year":2002,"claim":"Demonstrated CRKL activates Rac1 through DOCK GEFs and is genetically required for IFNα-dependent GAS transcription and Rap1 activation, extending the effector repertoire to actin-regulatory GTPases and confirming nuclear function in loss-of-function cells.","evidence":"DOCK2 co-IP and Rac1-GTP assays, CrkL-null MEF reporter and Rap1 assays with dominant-negative kinases","pmids":["12393632","11866427"],"confidence":"High","gaps":["Selectivity among DOCK family GEFs unclear","How one adaptor coordinates Rap1 vs Rac1 outputs spatially undefined"]},{"year":2003,"claim":"Showed Src-dependent, p130Cas-mediated recruitment of CRKL to focal adhesions activates Rac1/Cdc42 and is required for integrin-induced migration even when CrkII is present, establishing a non-redundant CRKL function in adhesion signaling.","evidence":"CrkL-null MEF rescue, forced-localization chimeras, GTPase and haptotaxis assays, ASAP1 pulldown/MS","pmids":["12665586","12522101"],"confidence":"High","gaps":["Basis for CrkL/CrkII functional non-equivalence at this stage unexplained","In vivo relevance of FA recruitment not yet shown"]},{"year":2006,"claim":"Established CRKL as a dosage-sensitive 22q11 gene whose loss phenocopies DiGeorge syndrome, acting in FGF8/FGFR and Tbx1/retinoic-acid pathways during neural crest and pharyngeal development.","evidence":"Crkl knockout and compound-heterozygous mouse genetics, Fgf8 stimulation/receptor binding, ERK assays, RA-pathway epistasis","pmids":["11242111","16399080","16399079"],"confidence":"High","gaps":["Direct molecular link between CRKL and RA signaling unresolved","Cell-autonomous vs non-autonomous neural crest roles not separated"]},{"year":2009,"claim":"Identified CRKL amplification as a lung cancer driver activating SOS1-Ras-ERK and Src-C3G-Rap1 and conferring EGFR-inhibitor resistance, and resolved the FGFR1 pY463 SH2 interaction structurally to explain CRKL's ~30-fold selectivity over CRK.","evidence":"shRNA/overexpression in NSCLC with pathway inhibitors, molecular dynamics and phosphopeptide binding affinity, Crkl-null ERK/growth assays","pmids":["22586683","19307307","19454690"],"confidence":"High","gaps":["Generality of CRKL amplification across tumor types unquantified","Structural model not validated by crystallography"]},{"year":2012,"claim":"Resolved by NMR why Bcr-Abl prefers CRKL over CrkII—CRKL forms a constitutive Abl complex with distinct domain regulation—providing the structural basis for CRKL's central role in CML.","evidence":"NMR structural analysis of CRKL and phospho-CRKL with binding-activity assays","pmids":["22581121"],"confidence":"High","gaps":["Full-length CRKL conformational dynamics not captured","Structural state in cells inferred, not directly observed"]},{"year":2010,"claim":"Established CRKL as functionally required for BCR-ABL leukemogenesis and as a tractable SH2-domain target, linking adaptor function to c-MYC and transformed growth.","evidence":"CrkL-null progenitor transplantation, SH2-blocking peptide, immunoprecipitation","pmids":["20807813"],"confidence":"High","gaps":["Therapeutic window of SH2 blockade untested","Downstream effectors beyond c-MYC not enumerated"]},{"year":2010,"claim":"Placed CRKL downstream of Reelin/Dab1 and MuSK/Dok-7 in vivo, establishing essential, partly redundant Crk/CrkL roles in cortical neuron positioning, dendritogenesis, and neuromuscular synapse formation.","evidence":"Tissue-specific Crk/CrkL knockouts, Dab1/Dok-7 phosphotyrosine mapping, Rap1 assays, NMJ and dendrite phenotyping","pmids":["15062102","18477607","21041412","18809728"],"confidence":"High","gaps":["Functional distinction between Crk and CrkL at synapses unresolved","Direct effectors at the NMJ not fully mapped"]},{"year":2014,"claim":"Demonstrated broad essential adaptor function via Crk/CrkL double knockout—loss of focal adhesions, stress fibers, and motility—and tissue-specific roles in podocyte and genitourinary development, showing overlapping but indispensable cytoskeletal regulation.","evidence":"Conditional double-knockout fibroblasts and podocytes, morphology/motility/lamellipodia assays, Crkl allelic series and RNA-seq","pmids":["24166500","24499776","25658046","26527617","28439006"],"confidence":"High","gaps":["Molecular reason single deletions are mild not fully explained","Direct transcriptional targets in GU development (e.g., Foxd1 regulation) mechanism unknown"]},{"year":2018,"claim":"Defined CRKL as a metastasis and EMT effector regulated by Zeb1/miR-200 and antagonized by SASH1, linking CRKL-driven Src/p130Cas-PI3K signaling to invasion and poor therapy response.","evidence":"miR-200 modulation, CRKL knockdown, in vivo metastasis models, SASH1 yeast two-hybrid/co-IP/MS, CRISPR epistasis, PTEN-null PI3K assays","pmids":["26728244","30480076","28723560"],"confidence":"High","gaps":["Whether CRKL's adaptor or scaffold activity drives EMT is unresolved","Context-dependence across tumor types not systematically tested"]},{"year":2024,"claim":"Uncovered a non-canonical CRKL function in immune evasion—stabilizing β-catenin by displacing Axin1 from APC to drive a VEGFα/CXCL1/neutrophil axis that confers anti-PD-1 resistance—extending CRKL biology beyond tyrosine-kinase adaptation.","evidence":"Pooled CRISPR screen, transgenic HCC mouse model, mass cytometry, co-IP, patient-derived tumor spheroids","pmids":["38403027"],"confidence":"High","gaps":["Structural basis of CRKL–APC/Axin1 competition unresolved","How tyrosine-adaptor functions relate to this β-catenin role unclear"]},{"year":null,"claim":"How CRKL's spatial and temporal switching among Rap1, Rac1/Cdc42, ERK, nuclear STAT5, and β-catenin outputs is selected in a given cell context remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No unified model linking domain occupancy to pathway choice","Mechanism of nuclear import undefined","Endogenous regulation of Y207 phosphorylation in normal physiology unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,10,11,18,33,49]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[10,33]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[18,24,32]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[30,33,49]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[11,22,33,42]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[26,36,37,45,53,57]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[1,42,58,59]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[32,40,43]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[18,24,32]}],"complexes":["BCR/ABL–CRKL–CBL complex","CRKL–STAT5 nuclear complex","CrkL–C3G complex"],"partners":["ABL1","BCR","CBL","RAPGEF1","BCAR1","STAT5","DOCK2","PIK3R1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"P46109","full_name":"Crk-like protein","aliases":[],"length_aa":303,"mass_kda":33.8,"function":"May mediate the transduction of intracellular signals","subcellular_location":"","url":"https://www.uniprot.org/uniprotkb/P46109/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CRKL","classification":"Common Essential","n_dependent_lines":901,"n_total_lines":1208,"dependency_fraction":0.7458609271523179},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CAPZB","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/CRKL","total_profiled":1310},"omim":[{"mim_id":"616912","title":"ENAH/VASP-LIKE PROTEIN; EVL","url":"https://www.omim.org/entry/616912"},{"mim_id":"616904","title":"DEDICATOR OF CYTOKINESIS 5; DOCK5","url":"https://www.omim.org/entry/616904"},{"mim_id":"615759","title":"KINASE D-INTERACTING SUBSTRATE, 220-KD; KIDINS220","url":"https://www.omim.org/entry/615759"},{"mim_id":"610285","title":"DOWNSTREAM OF TYROSINE KINASE 7; DOK7","url":"https://www.omim.org/entry/610285"},{"mim_id":"608541","title":"RHO GTPase-ACTIVATING PROTEIN 32; ARHGAP32","url":"https://www.omim.org/entry/608541"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/CRKL"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"P46109","domains":[{"cath_id":"3.30.505.10","chopping":"20-100","consensus_level":"high","plddt":83.2209,"start":20,"end":100},{"cath_id":"2.30.30.40","chopping":"125-180","consensus_level":"high","plddt":79.6532,"start":125,"end":180},{"cath_id":"2.30.30.40","chopping":"237-293","consensus_level":"high","plddt":77.6402,"start":237,"end":293}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/P46109","model_url":"https://alphafold.ebi.ac.uk/files/AF-P46109-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-P46109-F1-predicted_aligned_error_v6.png","plddt_mean":68.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CRKL","jax_strain_url":"https://www.jax.org/strain/search?query=CRKL"},"sequence":{"accession":"P46109","fasta_url":"https://rest.uniprot.org/uniprotkb/P46109.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/P46109/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/P46109"}},"corpus_meta":[{"pmid":"8632906","id":"PMC_8632906","title":"The proto-oncogene product p120CBL and the adaptor proteins CRKL and c-CRK link c-ABL, p190BCR/ABL and p210BCR/ABL to the phosphatidylinositol-3' kinase pathway.","date":"1996","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/8632906","citation_count":251,"is_preprint":false},{"pmid":"11242111","id":"PMC_11242111","title":"Mice lacking the homologue of the human 22q11.2 gene CRKL phenocopy neurocristopathies of DiGeorge syndrome.","date":"2001","source":"Nature genetics","url":"https://pubmed.ncbi.nlm.nih.gov/11242111","citation_count":238,"is_preprint":false},{"pmid":"19426560","id":"PMC_19426560","title":"Crk and CrkL adaptor proteins: networks for physiological and pathological signaling.","date":"2009","source":"Cell communication and signaling : CCS","url":"https://pubmed.ncbi.nlm.nih.gov/19426560","citation_count":226,"is_preprint":false},{"pmid":"8083188","id":"PMC_8083188","title":"Crkl is the major tyrosine-phosphorylated protein in neutrophils from patients with chronic myelogenous leukemia.","date":"1994","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8083188","citation_count":221,"is_preprint":false},{"pmid":"8361759","id":"PMC_8361759","title":"Isolation and chromosomal localization of CRKL, a human crk-like gene.","date":"1993","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/8361759","citation_count":214,"is_preprint":false},{"pmid":"7524758","id":"PMC_7524758","title":"Identification of CRKL as the constitutively phosphorylated 39-kD tyrosine phosphoprotein in chronic myelogenous leukemia cells.","date":"1994","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/7524758","citation_count":186,"is_preprint":false},{"pmid":"7521685","id":"PMC_7521685","title":"Tyrosine phosphorylation of CRKL in Philadelphia+ leukemia.","date":"1994","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/7521685","citation_count":185,"is_preprint":false},{"pmid":"15062102","id":"PMC_15062102","title":"Activation of a Dab1/CrkL/C3G/Rap1 pathway in Reelin-stimulated neurons.","date":"2004","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/15062102","citation_count":165,"is_preprint":false},{"pmid":"16399080","id":"PMC_16399080","title":"Dose-dependent interaction of Tbx1 and Crkl and locally aberrant RA signaling in a model of del22q11 syndrome.","date":"2006","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/16399080","citation_count":162,"is_preprint":false},{"pmid":"7545163","id":"PMC_7545163","title":"Crkl is complexed with tyrosine-phosphorylated Cbl in Ph-positive leukemia.","date":"1995","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7545163","citation_count":134,"is_preprint":false},{"pmid":"7493940","id":"PMC_7493940","title":"CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells.","date":"1995","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/7493940","citation_count":132,"is_preprint":false},{"pmid":"8798523","id":"PMC_8798523","title":"The CRKL adaptor protein transforms fibroblasts and functions in transformation by the BCR-ABL oncogene.","date":"1996","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8798523","citation_count":122,"is_preprint":false},{"pmid":"8168080","id":"PMC_8168080","title":"Cellular interactions of CRKL, and SH2-SH3 adaptor protein.","date":"1994","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/8168080","citation_count":122,"is_preprint":false},{"pmid":"22586683","id":"PMC_22586683","title":"Amplification of CRKL induces transformation and epidermal growth factor receptor inhibitor resistance in human non-small cell lung cancers.","date":"2011","source":"Cancer discovery","url":"https://pubmed.ncbi.nlm.nih.gov/22586683","citation_count":121,"is_preprint":false},{"pmid":"8810278","id":"PMC_8810278","title":"p130CAS forms a signaling complex with the adapter protein CRKL in hematopoietic cells transformed by the BCR/ABL oncogene.","date":"1996","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8810278","citation_count":117,"is_preprint":false},{"pmid":"10753869","id":"PMC_10753869","title":"Signaling of hepatocyte growth factor/scatter factor (HGF) to the small GTPase Rap1 via the large docking protein Gab1 and the adapter protein CRKL.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10753869","citation_count":116,"is_preprint":false},{"pmid":"16399079","id":"PMC_16399079","title":"Crkl deficiency disrupts Fgf8 signaling in a mouse model of 22q11 deletion syndromes.","date":"2006","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/16399079","citation_count":115,"is_preprint":false},{"pmid":"9872990","id":"PMC_9872990","title":"Activation of a CrkL-stat5 signaling complex by type I interferons.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9872990","citation_count":114,"is_preprint":false},{"pmid":"8621483","id":"PMC_8621483","title":"Sos, Vav, and C3G participate in B cell receptor-induced signaling pathways and differentially associate with Shc-Grb2, Crk, and Crk-L adaptors.","date":"1996","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/8621483","citation_count":111,"is_preprint":false},{"pmid":"9092574","id":"PMC_9092574","title":"Steel factor induces tyrosine phosphorylation of CRKL and binding of CRKL to a complex containing c-kit, phosphatidylinositol 3-kinase, and p120(CBL).","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9092574","citation_count":100,"is_preprint":false},{"pmid":"9593259","id":"PMC_9593259","title":"Role of the adapter protein CRKL in signal transduction of normal hematopoietic and BCR/ABL-transformed cells.","date":"1998","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/9593259","citation_count":98,"is_preprint":false},{"pmid":"10608804","id":"PMC_10608804","title":"The adapter protein Crkl links Cbl to C3G after integrin ligation and enhances cell migration.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10608804","citation_count":95,"is_preprint":false},{"pmid":"21041412","id":"PMC_21041412","title":"Dok-7 regulates neuromuscular synapse formation by recruiting Crk and Crk-L.","date":"2010","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/21041412","citation_count":94,"is_preprint":false},{"pmid":"9268367","id":"PMC_9268367","title":"Enhancement of guanine-nucleotide exchange activity of C3G for Rap1 by the expression of Crk, CrkL, and Grb2.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9268367","citation_count":91,"is_preprint":false},{"pmid":"18165234","id":"PMC_18165234","title":"Avian and 1918 Spanish influenza a virus NS1 proteins bind to Crk/CrkL Src homology 3 domains to activate host cell signaling.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/18165234","citation_count":87,"is_preprint":false},{"pmid":"9374471","id":"PMC_9374471","title":"The type I interferon receptor mediates tyrosine phosphorylation of the CrkL adaptor protein.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9374471","citation_count":84,"is_preprint":false},{"pmid":"18809728","id":"PMC_18809728","title":"The WAVE2 complex regulates T cell receptor signaling to integrins via Abl- and CrkL-C3G-mediated activation of Rap1.","date":"2008","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/18809728","citation_count":84,"is_preprint":false},{"pmid":"9162067","id":"PMC_9162067","title":"Differential signaling after beta1 integrin ligation is mediated through binding of CRKL to p120(CBL) and p110(HEF1).","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9162067","citation_count":84,"is_preprint":false},{"pmid":"12446789","id":"PMC_12446789","title":"Grit, a GTPase-activating protein for the Rho family, regulates neurite extension through association with the TrkA receptor and N-Shc and CrkL/Crk adapter molecules.","date":"2002","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12446789","citation_count":83,"is_preprint":false},{"pmid":"10339478","id":"PMC_10339478","title":"CrkL activates integrin-mediated hematopoietic cell adhesion through the guanine nucleotide exchange factor C3G.","date":"1999","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/10339478","citation_count":81,"is_preprint":false},{"pmid":"9891069","id":"PMC_9891069","title":"Interaction of hematopoietic progenitor kinase 1 with adapter proteins Crk and CrkL leads to synergistic activation of c-Jun N-terminal kinase.","date":"1999","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/9891069","citation_count":77,"is_preprint":false},{"pmid":"9053848","id":"PMC_9053848","title":"Tyrosine 207 in CRKL is the BCR/ABL phosphorylation site.","date":"1997","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9053848","citation_count":77,"is_preprint":false},{"pmid":"18477607","id":"PMC_18477607","title":"Reduction of Crk and CrkL expression blocks reelin-induced dendritogenesis.","date":"2008","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/18477607","citation_count":74,"is_preprint":false},{"pmid":"8978305","id":"PMC_8978305","title":"Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation.","date":"1997","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/8978305","citation_count":72,"is_preprint":false},{"pmid":"9591773","id":"PMC_9591773","title":"Development of highly selective SH3 binding peptides for Crk and CRKL which disrupt Crk-complexes with DOCK180, SoS and C3G.","date":"1998","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9591773","citation_count":68,"is_preprint":false},{"pmid":"19966867","id":"PMC_19966867","title":"Genomic and functional analysis identifies CRKL as an oncogene amplified in lung cancer.","date":"2009","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/19966867","citation_count":66,"is_preprint":false},{"pmid":"9710592","id":"PMC_9710592","title":"Structural requirements for function of the Crkl adapter protein in fibroblasts and hematopoietic cells.","date":"1998","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/9710592","citation_count":65,"is_preprint":false},{"pmid":"12393632","id":"PMC_12393632","title":"DOCK2 associates with CrkL and regulates Rac1 in human leukemia cell lines.","date":"2002","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/12393632","citation_count":63,"is_preprint":false},{"pmid":"9195915","id":"PMC_9195915","title":"Interactions of CBL with BCR-ABL and CRKL in BCR-ABL-transformed myeloid cells.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9195915","citation_count":63,"is_preprint":false},{"pmid":"12665586","id":"PMC_12665586","title":"Translocation of CrkL to focal adhesions mediates integrin-induced migration downstream of Src family kinases.","date":"2003","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/12665586","citation_count":63,"is_preprint":false},{"pmid":"18403635","id":"PMC_18403635","title":"MUC1 initiates Src-CrkL-Rac1/Cdc42-mediated actin cytoskeletal protrusive motility after ligating intercellular adhesion molecule-1.","date":"2008","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/18403635","citation_count":61,"is_preprint":false},{"pmid":"26728244","id":"PMC_26728244","title":"The microRNA-200/Zeb1 axis regulates ECM-dependent β1-integrin/FAK signaling, cancer cell invasion and metastasis through CRKL.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26728244","citation_count":58,"is_preprint":false},{"pmid":"25658046","id":"PMC_25658046","title":"Mouse and human CRKL is dosage sensitive for cardiac outflow tract formation.","date":"2015","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/25658046","citation_count":58,"is_preprint":false},{"pmid":"29344280","id":"PMC_29344280","title":"MiR-378 and MiR-1827 Regulate Tumor Invasion, Migration and Angiogenesis in Human Lung Adenocarcinoma by Targeting RBX1 and CRKL, Respectively.","date":"2018","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/29344280","citation_count":56,"is_preprint":false},{"pmid":"38403027","id":"PMC_38403027","title":"CRKL dictates anti-PD-1 resistance by mediating tumor-associated neutrophil infiltration in hepatocellular carcinoma.","date":"2024","source":"Journal of hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/38403027","citation_count":54,"is_preprint":false},{"pmid":"9461587","id":"PMC_9461587","title":"Interleukin-2 stimulation induces tyrosine phosphorylation of p120-Cbl and CrkL and formation of multimolecular signaling complexes in T lymphocytes and natural killer cells.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9461587","citation_count":51,"is_preprint":false},{"pmid":"22581121","id":"PMC_22581121","title":"Domain organization differences explain Bcr-Abl's preference for CrkL over CrkII.","date":"2012","source":"Nature chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/22581121","citation_count":51,"is_preprint":false},{"pmid":"10514505","id":"PMC_10514505","title":"CrkL mediates Ras-dependent activation of the Raf/ERK pathway through the guanine nucleotide exchange factor C3G in hematopoietic cells stimulated with erythropoietin or interleukin-3.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10514505","citation_count":51,"is_preprint":false},{"pmid":"26044596","id":"PMC_26044596","title":"CrkL meditates CCL20/CCR6-induced EMT in gastric cancer.","date":"2015","source":"Cytokine","url":"https://pubmed.ncbi.nlm.nih.gov/26044596","citation_count":50,"is_preprint":false},{"pmid":"19454690","id":"PMC_19454690","title":"PI3K links NKG2D signaling to a CrkL pathway involved in natural killer cell adhesion, polarity, and granule secretion.","date":"2009","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/19454690","citation_count":49,"is_preprint":false},{"pmid":"29403024","id":"PMC_29403024","title":"miR-429 suppresses tumor migration and invasion by targeting CRKL in hepatocellular carcinoma via inhibiting Raf/MEK/ERK pathway and epithelial-mesenchymal transition.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29403024","citation_count":49,"is_preprint":false},{"pmid":"12522101","id":"PMC_12522101","title":"CrkL directs ASAP1 to peripheral focal adhesions.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12522101","citation_count":49,"is_preprint":false},{"pmid":"28887306","id":"PMC_28887306","title":"The long noncoding RNA PCAT-1 links the microRNA miR-215 to oncogene CRKL-mediated signaling in hepatocellular carcinoma.","date":"2017","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/28887306","citation_count":48,"is_preprint":false},{"pmid":"10428508","id":"PMC_10428508","title":"CrkL and CrkII participate in the generation of the growth inhibitory effects of interferons on primary hematopoietic progenitors.","date":"1999","source":"Experimental hematology","url":"https://pubmed.ncbi.nlm.nih.gov/10428508","citation_count":48,"is_preprint":false},{"pmid":"11031258","id":"PMC_11031258","title":"SHIP1, an SH2 domain containing polyinositol-5-phosphatase, regulates migration through two critical tyrosine residues and forms a novel signaling complex with DOK1 and CRKL.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11031258","citation_count":47,"is_preprint":false},{"pmid":"9405482","id":"PMC_9405482","title":"BCR/ABL-induced leukemogenesis causes phosphorylation of Hef1 and its association with Crkl.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9405482","citation_count":46,"is_preprint":false},{"pmid":"11245441","id":"PMC_11245441","title":"Crkl enhances leukemogenesis in BCR/ABL P190 transgenic mice.","date":"2001","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/11245441","citation_count":44,"is_preprint":false},{"pmid":"9344843","id":"PMC_9344843","title":"Erythropoietin and IL-3 induce tyrosine phosphorylation of CrkL and its association with Shc, SHP-2, and Cbl in hematopoietic cells.","date":"1997","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/9344843","citation_count":43,"is_preprint":false},{"pmid":"10720695","id":"PMC_10720695","title":"CrkL functions as a nuclear adaptor and transcriptional activator in Bcr-Abl-expressing cells.","date":"2000","source":"Experimental hematology","url":"https://pubmed.ncbi.nlm.nih.gov/10720695","citation_count":43,"is_preprint":false},{"pmid":"24055140","id":"PMC_24055140","title":"CRKL promotes cell proliferation in gastric cancer and is negatively regulated by miR-126.","date":"2013","source":"Chemico-biological interactions","url":"https://pubmed.ncbi.nlm.nih.gov/24055140","citation_count":42,"is_preprint":false},{"pmid":"23318429","id":"PMC_23318429","title":"Loss-of-function screen in rhabdomyosarcoma identifies CRKL-YES as a critical signal for tumor growth.","date":"2013","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/23318429","citation_count":42,"is_preprint":false},{"pmid":"29155146","id":"PMC_29155146","title":"HDAC inhibitor suppresses proliferation and invasion of breast cancer cells through regulation of miR-200c targeting CRKL.","date":"2017","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29155146","citation_count":41,"is_preprint":false},{"pmid":"33801580","id":"PMC_33801580","title":"Crk and CrkL as Therapeutic Targets for Cancer Treatment.","date":"2021","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/33801580","citation_count":40,"is_preprint":false},{"pmid":"24664993","id":"PMC_24664993","title":"CRKL promotes lung cancer cell invasion through ERK-MMP9 pathway.","date":"2014","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/24664993","citation_count":40,"is_preprint":false},{"pmid":"11313252","id":"PMC_11313252","title":"CrkL is an adapter for Wiskott-Aldrich syndrome protein and Syk.","date":"2001","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/11313252","citation_count":40,"is_preprint":false},{"pmid":"30480076","id":"PMC_30480076","title":"The Tumor Suppressor SASH1 Interacts With the Signal Adaptor CRKL to Inhibit Epithelial-Mesenchymal Transition and Metastasis in Colorectal Cancer.","date":"2018","source":"Cellular and molecular gastroenterology and hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/30480076","citation_count":40,"is_preprint":false},{"pmid":"28723560","id":"PMC_28723560","title":"CRKL Mediates p110β-Dependent PI3K Signaling in PTEN-Deficient Cancer Cells.","date":"2017","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/28723560","citation_count":39,"is_preprint":false},{"pmid":"9614078","id":"PMC_9614078","title":"Interplay of the proto-oncogene proteins CrkL and CrkII in insulin-like growth factor-I receptor-mediated signal transduction.","date":"1998","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9614078","citation_count":39,"is_preprint":false},{"pmid":"28439006","id":"PMC_28439006","title":"Murine model indicates 22q11.2 signaling adaptor CRKL is a dosage-sensitive regulator of genitourinary development.","date":"2017","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/28439006","citation_count":39,"is_preprint":false},{"pmid":"27133071","id":"PMC_27133071","title":"The effects of Micro-429 on inhibition of cervical cancer cells through targeting ZEB1 and CRKL.","date":"2016","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/27133071","citation_count":38,"is_preprint":false},{"pmid":"24499776","id":"PMC_24499776","title":"Crk1/2 and CrkL form a hetero-oligomer and functionally complement each other during podocyte morphogenesis.","date":"2014","source":"Kidney international","url":"https://pubmed.ncbi.nlm.nih.gov/24499776","citation_count":36,"is_preprint":false},{"pmid":"11389015","id":"PMC_11389015","title":"The adapter protein CrkL associates with CD34.","date":"2001","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/11389015","citation_count":36,"is_preprint":false},{"pmid":"11443118","id":"PMC_11443118","title":"CrkL is recruited through its SH2 domain to the erythropoietin receptor and plays a role in Lyn-mediated receptor signaling.","date":"2001","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11443118","citation_count":36,"is_preprint":false},{"pmid":"9067577","id":"PMC_9067577","title":"The BCR/ABL oncogene alters interaction of the adapter proteins CRKL and CRK with cellular proteins.","date":"1997","source":"Leukemia","url":"https://pubmed.ncbi.nlm.nih.gov/9067577","citation_count":36,"is_preprint":false},{"pmid":"7478571","id":"PMC_7478571","title":"Tyrosine phosphorylation of murine Crkl.","date":"1995","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/7478571","citation_count":35,"is_preprint":false},{"pmid":"9498705","id":"PMC_9498705","title":"Association of the Cas-like molecule HEF1 with CrkL following integrin and antigen receptor signaling in human B-cells: potential relevance to neoplastic lymphohematopoietic cells.","date":"1997","source":"Leukemia & lymphoma","url":"https://pubmed.ncbi.nlm.nih.gov/9498705","citation_count":35,"is_preprint":false},{"pmid":"11167825","id":"PMC_11167825","title":"Engagement of the CrkL adaptor in interferon alpha signalling in BCR-ABL-expressing cells.","date":"2001","source":"British journal of haematology","url":"https://pubmed.ncbi.nlm.nih.gov/11167825","citation_count":34,"is_preprint":false},{"pmid":"24166500","id":"PMC_24166500","title":"Essential roles of Crk and CrkL in fibroblast structure and motility.","date":"2013","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/24166500","citation_count":32,"is_preprint":false},{"pmid":"11866427","id":"PMC_11866427","title":"The CrkL adapter protein is required for type I interferon-dependent gene transcription and activation of the small G-protein Rap1.","date":"2002","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/11866427","citation_count":30,"is_preprint":false},{"pmid":"20807813","id":"PMC_20807813","title":"A specific need for CRKL in p210BCR-ABL-induced transformation of mouse hematopoietic progenitors.","date":"2010","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/20807813","citation_count":30,"is_preprint":false},{"pmid":"26527617","id":"PMC_26527617","title":"Sorbs1 and -2 Interact with CrkL and Are Required for Acetylcholine Receptor Cluster Formation.","date":"2015","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/26527617","citation_count":29,"is_preprint":false},{"pmid":"31133010","id":"PMC_31133010","title":"CRKL regulates alternative splicing of cancer-related genes in cervical cancer samples and HeLa cell.","date":"2019","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/31133010","citation_count":29,"is_preprint":false},{"pmid":"9199202","id":"PMC_9199202","title":"Inhibition of Grb2 and Crkl proteins results in growth inhibition of Philadelphia chromosome positive leukemic cells.","date":"1997","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/9199202","citation_count":29,"is_preprint":false},{"pmid":"10362355","id":"PMC_10362355","title":"Involvement of the adapter protein CRKL in integrin-mediated adhesion.","date":"1999","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/10362355","citation_count":28,"is_preprint":false},{"pmid":"9845531","id":"PMC_9845531","title":"Thrombopoietin induces association of Crkl with STAT5 but not STAT3 in human platelets.","date":"1998","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/9845531","citation_count":28,"is_preprint":false},{"pmid":"26079153","id":"PMC_26079153","title":"CRKL oncogene is downregulated by p53 through miR-200s.","date":"2015","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/26079153","citation_count":27,"is_preprint":false},{"pmid":"23686806","id":"PMC_23686806","title":"Overexpression of CRKL correlates with malignant cell proliferation in breast cancer.","date":"2013","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/23686806","citation_count":27,"is_preprint":false},{"pmid":"9346905","id":"PMC_9346905","title":"In vivo regulation of CrkII and CrkL proto-oncogenes in the uterus by insulin-like growth factor-I. Differential effects on tyrosine phosphorylation and association with paxillin.","date":"1997","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9346905","citation_count":27,"is_preprint":false},{"pmid":"9988765","id":"PMC_9988765","title":"Activation of nuclear factor of activated T cells-(NFAT) and activating protein 1 (AP-1) by oncogenic 70Z Cbl requires an intact phosphotyrosine binding domain but not Crk(L) or p85 phosphatidylinositol 3-kinase association.","date":"1999","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9988765","citation_count":27,"is_preprint":false},{"pmid":"31706051","id":"PMC_31706051","title":"MicroRNA-429 inhibits bone metastasis in breast cancer by regulating CrkL and MMP-9.","date":"2019","source":"Bone","url":"https://pubmed.ncbi.nlm.nih.gov/31706051","citation_count":26,"is_preprint":false},{"pmid":"23959425","id":"PMC_23959425","title":"CrkL efficiently mediates cell proliferation, migration, and invasion induced by TGF-β pathway in glioblastoma.","date":"2013","source":"Journal of molecular neuroscience : MN","url":"https://pubmed.ncbi.nlm.nih.gov/23959425","citation_count":25,"is_preprint":false},{"pmid":"32280542","id":"PMC_32280542","title":"Bidirectional interaction of lncRNA AFAP1-AS1 and CRKL accelerates the proliferative and metastatic abilities of hepatocarcinoma cells.","date":"2020","source":"Journal of advanced research","url":"https://pubmed.ncbi.nlm.nih.gov/32280542","citation_count":25,"is_preprint":false},{"pmid":"33094104","id":"PMC_33094104","title":"miR-124-3p Suppresses the Invasiveness and Metastasis of Hepatocarcinoma Cells via Targeting CRKL.","date":"2020","source":"Frontiers in molecular biosciences","url":"https://pubmed.ncbi.nlm.nih.gov/33094104","citation_count":25,"is_preprint":false},{"pmid":"19307307","id":"PMC_19307307","title":"Structural and functional basis of a role for CRKL in a fibroblast growth factor 8-induced feed-forward loop.","date":"2009","source":"Molecular and cellular biology","url":"https://pubmed.ncbi.nlm.nih.gov/19307307","citation_count":25,"is_preprint":false},{"pmid":"12082638","id":"PMC_12082638","title":"BCR/ABL P190 transgenic mice develop leukemia in the absence of Crkl.","date":"2002","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12082638","citation_count":25,"is_preprint":false},{"pmid":"10926930","id":"PMC_10926930","title":"Engagement of the CrkL adapter in interleukin-5 signaling in eosinophils.","date":"2000","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/10926930","citation_count":24,"is_preprint":false},{"pmid":"22244889","id":"PMC_22244889","title":"CRKL plays a pivotal role in tumorigenesis of head and neck squamous cell carcinoma through the regulation of cell adhesion.","date":"2012","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/22244889","citation_count":24,"is_preprint":false},{"pmid":"24145029","id":"PMC_24145029","title":"Escherichia coli virulence protein NleH1 interaction with the v-Crk sarcoma virus CT10 oncogene-like protein (CRKL) governs NleH1 inhibition of the ribosomal protein S3 (RPS3)/nuclear factor κB (NF-κB) pathway.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24145029","citation_count":24,"is_preprint":false},{"pmid":"16781119","id":"PMC_16781119","title":"CrkL plays a role in SDF-1-induced activation of the Raf-1/MEK/Erk pathway through Ras and Rac to mediate chemotactic signaling in hematopoietic cells.","date":"2006","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/16781119","citation_count":24,"is_preprint":false},{"pmid":"21319228","id":"PMC_21319228","title":"Crk and CrkL present with different expression and significance in epithelial ovarian carcinoma.","date":"2011","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/21319228","citation_count":23,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":51664,"output_tokens":14187,"usd":0.183898,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":26300,"output_tokens":6054,"usd":0.141425,"stage2_stop_reason":"end_turn"},"total_usd":0.325323,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1993,\n      \"finding\": \"CRKL was identified as a novel gene on chromosome 22q11 encoding a 303 amino acid protein with one SH2 and two SH3 domains, structurally related to but distinct from c-CRK (v-crk oncogene homolog), predicted to function as a signal transduction adaptor.\",\n      \"method\": \"cDNA cloning, sequence analysis\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — direct sequencing and domain identification in a single foundational study; no functional validation beyond sequence\",\n      \"pmids\": [\"8361759\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"CRKL is the major tyrosine-phosphorylated protein (39 kDa) in CML neutrophils and cell lines expressing p210BCR-ABL, and a direct interaction between CRKL and ABL was demonstrated by yeast two-hybrid screen.\",\n      \"method\": \"Anti-phosphotyrosine immunoblotting, protein purification, microsequencing, yeast two-hybrid\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — independently replicated by multiple labs (PMID 8083188, 7524758, 7521685) with orthogonal methods\",\n      \"pmids\": [\"8083188\", \"7524758\", \"7521685\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"CRKL is a substrate for the p210BCR/ABL and p145ABL kinases in vivo; BCR/ABL and ABL co-immunoprecipitate with CRKL, forming stable complexes; mSOS1 also co-immunoprecipitates with CRKL, establishing a putative signaling pathway from BCR/ABL through CRKL to SOS.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP and kinase assay, replicated across multiple subsequent studies\",\n      \"pmids\": [\"8168080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"The CRKL SH2 domain binds specifically to tyrosine-phosphorylated CBL in Ph-positive leukemia cells; CRKL SH3 domains bind BCR/ABL but not CBL; a trimolecular complex of BCR/ABL–CRKL–CBL exists in Ph-positive cells.\",\n      \"method\": \"GST fusion protein pulldown, co-immunoprecipitation, domain mapping\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP with domain mapping, replicated in multiple cell contexts\",\n      \"pmids\": [\"7545163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"CRKL SH3 domains bind c-ABL and p210BCR/ABL; CRKL SH2 domain binds paxillin (a focal adhesion protein) at tyrosines 31 and 118 (but not 181), physically linking p210BCR/ABL to paxillin in CML cells.\",\n      \"method\": \"GST-CRKL fusion protein binding assay, site-directed mutagenesis, co-immunoprecipitation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro binding with mutagenesis identifying specific phosphotyrosine residues, single lab\",\n      \"pmids\": [\"7493940\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"CRKL SH2 domains bind p120CBL; CRKL and c-CRK SH3 domains bind BCR/ABL and c-ABL; CRKL participates in multimeric complexes including p120CBL, PI3K, and BCR/ABL, linking BCR/ABL to the PI3K pathway.\",\n      \"method\": \"In vitro binding studies, co-immunoprecipitation, PI3K lipid kinase activity assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro domain binding plus functional PI3K activity assay in co-precipitates, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"8632906\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"CRKL overexpression activates RAS and JUN kinase signaling pathways and transforms fibroblasts in a RAS-dependent manner. CRKL contributes to BCR-ABL fibroblast transformation alongside GRB2; deletion of both CRKL and GRB2 binding sites reduced transforming activity 15-fold.\",\n      \"method\": \"Fibroblast transformation assay, RAS activation assay, BCR-ABL deletion mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional transformation assay with domain deletion mutants and epistasis, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"8798523\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1996,\n      \"finding\": \"CRKL SH2 domain binds tyrosine-phosphorylated p130CAS in BCR/ABL-transformed cells; p130CAS is tyrosine-phosphorylated and constitutively associated with CRKL in CML cells; BCR/ABL disrupts the normal interaction between p130CAS and tensin but not p130CAS–FAK or p130CAS–paxillin.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP in cell lines and patient samples, single lab\",\n      \"pmids\": [\"8810278\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Tyrosine 207 in CRKL (between the two SH3 domains) is the major BCR/ABL phosphorylation site in vivo; Y207F mutation eliminates tyrosine phosphorylation; phosphorylation at Y207 provides a binding site for the CRKL SH2 domain itself (autoinhibitory interaction) but does not alter stoichiometry of SOS or C3G complex formation.\",\n      \"method\": \"In vitro kinase assay, site-directed mutagenesis, tryptic phosphopeptide mapping\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay with mutagenesis identifying specific phosphorylation site, single lab\",\n      \"pmids\": [\"9053848\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"CRKL N-terminal SH3 domain directly binds BCR/ABL at a proline-rich C-terminal region of ABL; however, deletion of this proline-rich region did not impair BCR/ABL factor-independent transformation of myeloid cells, and CRKL still became phosphorylated via indirect interaction with the mutant BCR/ABL.\",\n      \"method\": \"Yeast two-hybrid, gel overlay, co-immunoprecipitation, factor-independent growth assay\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding mapping with negative functional result for direct interaction requirement, single lab\",\n      \"pmids\": [\"8978305\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"After beta1 integrin ligation, CRKL SH2 domain binds tyrosine-phosphorylated p120CBL in megakaryocytic MO7e cells and p110HEF1 in lymphoid H9 cells; CRKL is constitutively complexed to C3G, SOS, and c-ABL via its SH3 domains regardless of integrin ligation, enabling cell-type-specific signaling complexes.\",\n      \"method\": \"Co-immunoprecipitation, domain-specific binding assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP in two distinct cell lines with domain specificity mapping, single lab\",\n      \"pmids\": [\"9162067\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"CrkL N-terminal SH3 domain binds C3G, and co-expression of CrkL with C3G in Cos1 cells significantly increased GTP/GDP ratio on Rap1; CrkL enhances C3G-mediated Rap1 activation primarily by membrane recruitment of C3G, requiring both SH2 and SH3 domains (SH2 requirement compensable by farnesylation signal).\",\n      \"method\": \"Co-expression/overexpression, GTP-binding assay, farnesylation rescue experiment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional GTPase assay with mechanistic dissection using domain mutants and rescue, single lab\",\n      \"pmids\": [\"9268367\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"BCR/ABL-induced leukemogenesis causes tyrosine phosphorylation of Hef1/Cas-L and its association with the CRKL SH2 domain; P190BCR/ABL, CRKL, and Hef1 or p120CBL form complexes in leukemic tissues, supporting a model where CRKL mediates BCR/ABL signaling to beta1-integrin pathway components.\",\n      \"method\": \"Co-immunoprecipitation, GST-domain binding in transgenic mouse leukemic tissue\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP in primary leukemic tissue but single lab, single method\",\n      \"pmids\": [\"9405482\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Type I interferon (IFNα) induces rapid tyrosine phosphorylation of CRKL; this phosphorylation is regulated by the IFN receptor-associated Tyk-2 kinase, as shown by Tyk-2 kinase activity in anti-CrkL immunoprecipitates and IFNα-dependent association of CrkL with Tyk-2.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP with kinase activity assay, single lab\",\n      \"pmids\": [\"9374471\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"CBL binds directly to the SH2 domain of BCR-ABL when CBL is tyrosine-phosphorylated; CRKL mediates an indirect complex between CBL and BCR-ABL, since deletion of the BCR-ABL SH2 domain did not abolish CBL–BCR-ABL complex formation.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP with domain deletion mutants, single lab\",\n      \"pmids\": [\"9195915\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"Steel factor (SF) activation of c-Kit induces CRKL tyrosine phosphorylation; CRKL co-precipitates with c-Kit through CRKL SH3 domains (not SH2) as part of a larger complex containing p85PI3K and p120CBL; CRKL binds directly to p85PI3K in vitro via its SH3 domain.\",\n      \"method\": \"Co-immunoprecipitation, Far Western blotting with GST-SH3 fusion protein\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro direct binding assay plus in vivo co-IP, single lab\",\n      \"pmids\": [\"9092574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"CRKL Y207F mutation abolishes all in vivo tyrosine phosphorylation and paradoxically enhances CRKL function (complex formation with SH2-binding proteins, JNK signaling, fibroblast transformation), indicating Y207 phosphorylation acts as a negative regulatory site; both SH2 and N-terminal SH3 domains are essential for CRKL biological activity in fibroblasts (transformation) and hematopoietic cells (adhesion).\",\n      \"method\": \"Tryptic phosphopeptide mapping, site-directed mutagenesis, transformation assay, JNK kinase assay, adhesion assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — mutagenesis with multiple functional readouts, single lab\",\n      \"pmids\": [\"9710592\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Highly selective peptides binding exclusively to the N-terminal SH3 domains of CrkL and Crk (but not Grb2 SH3 or other SH3 domains) were developed and shown to disrupt pre-existing Crk complexes with DOCK180, SOS, and C3G in a concentration-dependent manner.\",\n      \"method\": \"GST-fusion peptide pulldown, in-solution precipitation with biotinylated peptides, SH3 domain binding specificity panel\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain specificity demonstrated with broad panel of SH3 domains and complex disruption assays, single lab\",\n      \"pmids\": [\"9591773\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"IFNα-phosphorylated STAT5 acts as a docking site for the CRKL SH2 domain; CRKL and STAT5 form a complex that translocates to the nucleus and binds the TTCTAGGAA palindromic element in promoters of a subset of IFN-stimulated genes, establishing CRKL as a nuclear adapter regulating gene transcription.\",\n      \"method\": \"Co-immunoprecipitation, EMSA (electrophoretic mobility shift assay), nuclear fractionation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus functional EMSA with DNA binding, nuclear translocation confirmed, single lab\",\n      \"pmids\": [\"9872990\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"HPK1 (hematopoietic progenitor kinase 1) interacts with CRKL in vitro and in vivo via CRKL SH3 domain binding to proline-rich motifs in HPK1; CRKL synergizes with HPK1 to activate JNK; HPK1 phosphorylates CRKL mainly on serine/threonine residues in vitro.\",\n      \"method\": \"Co-immunoprecipitation, in vitro kinase assay, JNK activation assay, dominant-negative epistasis\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro kinase assay plus in vivo co-IP with functional JNK readout, single lab\",\n      \"pmids\": [\"9891069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"CRKL overexpression enhances spontaneous cell migration requiring both SH2 and N-terminal SH3 domains; after integrin cross-linking, full-length but not ΔSH2 CRKL co-precipitates tyrosine-phosphorylated CBL; the major CRKL SH3-binding protein in hematopoietic Ba/F3 cells is C3G; overexpression of C3G also enhances migration, suggesting a CBL–CRKL–C3G complex in migration signaling.\",\n      \"method\": \"Transwell migration assay, co-immunoprecipitation with domain deletion mutants, FACS-sorted GFP-tagged constructs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional migration assay with domain mutants plus protein interaction mapping, single lab\",\n      \"pmids\": [\"10608804\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"CrkL overexpression enhances hematopoietic cell adhesion to fibronectin via VLA-4 and VLA-5 integrins without changing integrin surface expression; the N-terminal SH3 domain (C3G binding) is critical, while C-terminal SH3 and Y207 are dispensable; C3G overexpression also increases adhesion and C3G mutant lacking the GEF domain blocks CrkL-induced adhesion, establishing a CrkL–C3G pathway activating integrins.\",\n      \"method\": \"Adhesion assay, domain-deletion mutant analysis, dominant negative C3G\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional assay with systematic domain deletion and epistasis using dominant-negative C3G, single lab\",\n      \"pmids\": [\"10339478\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"CrkL overexpression enhances Epo/IL-3-induced ERK1/2 activation (augmented and prolonged) via a Ras-dependent mechanism through C3G; CrkL mutants lacking SH2 or N-terminal SH3 domains inhibit Epo-induced ERK2 activation; CrkL also modestly activates JNK.\",\n      \"method\": \"Elk-1 reporter assay, ERK kinase assay, Ras-GTP pulldown, dominant-negative Ras\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional signaling assay with domain mutants and epistasis, single lab\",\n      \"pmids\": [\"10514505\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"HGF stimulation induces association of CRKL (via SH2 domain) with phosphorylated YXXP motifs in the docking protein GAB1; CRKL's N-terminal SH3 domain then binds C3G, activating Rap1; CRKL–C3G interaction is required for HGF-induced Rap1 activation and contributes to reduced cell adhesion/migration.\",\n      \"method\": \"Co-immunoprecipitation, dominant-negative C3G transfection, Rap1-GTP assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain mapping plus functional Rap1 GTPase assay and rescue experiments, single lab\",\n      \"pmids\": [\"10753869\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"In BCR-ABL-expressing cells, CRKL interacts with tyrosine-phosphorylated STAT5 and is found in the nucleus, detectable in a STAT5/DNA complex; CRKL increases transcriptional activation from a STAT-responsive reporter, functioning as a nuclear adaptor that associates with and activates STAT proteins.\",\n      \"method\": \"Co-immunoprecipitation, indirect immunofluorescence, EMSA supershift, luciferase reporter assay\",\n      \"journal\": \"Experimental hematology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus functional transcriptional assay with nuclear localization, single lab\",\n      \"pmids\": [\"10720695\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"SHIP1 forms a signaling complex with DOK1, PI3K, and CRKL in BCR/ABL-transformed cells; DOK1 binds SHIP1 directly via its PTB domain; CRKL directly binds SHIP1 via the CRKL SH2 domain; SHIP1 tyrosines 917 and 1020 mediate interactions with DOK1; expression of tyrosine-mutant SHIP1 fails to alter migration.\",\n      \"method\": \"Co-immunoprecipitation, direct binding assays, migration assay with tyrosine mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct binding mapped to specific tyrosine residues with functional migration readout, single lab\",\n      \"pmids\": [\"11031258\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Crkl-null mice exhibit defects in cranial/cardiac neural crest derivatives (ganglia, aortic arch arteries, cardiac outflow tract, thymus, parathyroid, craniofacial structures) phenocopying DiGeorge syndrome; neural crest migration and early expansion are unaffected, indicating an essential role for Crkl in neural crest function, differentiation, and/or survival.\",\n      \"method\": \"Targeted gene knockout (null mutation), embryo phenotypic analysis\",\n      \"journal\": \"Nature genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean knockout with defined tissue-specific phenotype, multiple organ systems analyzed\",\n      \"pmids\": [\"11242111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"CrkL C-terminal SH3 domain binds CD34 at a membrane-proximal region of the CD34 intracellular tail; this interaction is specific to CrkL (not CrkII); CrkL co-immunoprecipitates with CD34 in vivo.\",\n      \"method\": \"GST-fusion protein pulldown, co-immunoprecipitation, domain mapping with deletion constructs\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — GST pulldown plus in vivo co-IP confirming C-terminal SH3-mediated interaction, single lab\",\n      \"pmids\": [\"11389015\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"CrkL binds WASP through its SH3 domain (binding unaffected by WASP tyrosine phosphorylation) and also associates with Syk tyrosine kinase via both SH2 and SH3 domains; CrkL immunoprecipitates contain kinase-active Syk; CrkL acts as a molecular adapter linking WASP and Syk.\",\n      \"method\": \"Co-immunoprecipitation, GST fusion protein pulldown, in vitro kinase assay\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain dissection and functional kinase assay, single lab\",\n      \"pmids\": [\"11313252\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"CRKL overexpression in BCR/ABL transgenic mice enhances constitutive C3G complex formation, increases integrin-based macrophage motility, and markedly accelerates BCR/ABL-induced leukemia/lymphoma onset; Rap1 is activated in CRKL-overexpressing metastatic tumors.\",\n      \"method\": \"Transgenic mouse model, co-immunoprecipitation, Rap1 activation assay, migration assay\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo transgenic model with mechanistic co-IP and GTPase assays, single lab\",\n      \"pmids\": [\"11245441\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"DOCK2 binds to the N-terminal SH3 domain of CrkL via two separate regions; CrkL and DOCK2 co-localize with F-actin; CrkL-induced Rac1 activation is inhibited by a DOCK2 mutant; DOCK2 associates with Vav in Jurkat cells, revealing a CrkL–DOCK2–Vav complex regulating Rac1.\",\n      \"method\": \"Co-immunoprecipitation (in vitro and in vivo), immunocytochemistry, Rac1-GTP assay, cell attachment assay\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo binding with functional Rac1 GTPase assay, single lab\",\n      \"pmids\": [\"12393632\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Grit (a RhoGAP for Rho/Rac/Cdc42) directly interacts with TrkA and is also recruited to activated TrkA via binding to N-Shc and CrkL/Crk adapters; overexpression of the TrkA-binding region of Grit inhibited NGF-induced neurite elongation.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, overexpression inhibition assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP identifying complex, functional inhibition assay, single lab\",\n      \"pmids\": [\"12446789\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"CrkL is required for type I IFN-α-dependent gene transcription via GAS elements (through CrkL:Stat5 complexes); IFN-α-induced Rap1 activation is defective in CrkL-null MEFs; this Rap1 activation requires the N-terminal SH3 domain of CrkL and both Tyk-2 and Jak-1 kinases.\",\n      \"method\": \"CrkL-/- mouse embryonic fibroblasts, reporter gene assay, Rap1-GTP assay, dominant-negative kinase constructs\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function in null cells with multiple functional readouts and domain rescue, single lab\",\n      \"pmids\": [\"11866427\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Src triggers translocation of CrkL to focal adhesions in a manner dependent on p130Cas; forced localization of CRKL to focal adhesions activates Rac1 and Cdc42 and rescues haptotaxis defects of Src/Yes/Fyn triple-knockout MEFs; CrkL co-recruits Dock1 to focal adhesions for Rac1 activation; MEFs lacking CrkL show impaired integrin-induced migration despite expressing CrkII.\",\n      \"method\": \"MEF knockout/rescue, live-cell imaging, Rac1/Cdc42 GTPase assay, haptotaxis assay, forced-localization chimeric construct\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic loss-of-function with rescue, GTPase assay, and mechanistic forced-localization experiment, single lab\",\n      \"pmids\": [\"12665586\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"CrkL directs ASAP1 (an Arf-GAP) to peripheral focal adhesions; CrkL N-terminal SH3 domain binds ASAP1 in platelets (identified by pulldown/mass spectrometry); co-expression of wild-type CrkL (but not SH2-mutant CrkL, which cannot localize to FAs) recruits ASAP1 to focal adhesions.\",\n      \"method\": \"Pull-down assay followed by mass spectrometry, co-expression in COS7 cells, immunofluorescence\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pulldown/MS identification with functional localization experiment using SH2 mutant, single lab\",\n      \"pmids\": [\"12522101\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Reelin stimulation causes CrkL (and CrkI/II) to bind tyrosine-phosphorylated Dab1 at Y220 and Y232 (critical for cortical neuron positioning); CrkL also binds C3G; Reelin stimulates tyrosine phosphorylation of C3G and activates Rap1, establishing a Dab1–CrkL–C3G–Rap1 pathway downstream of Reelin.\",\n      \"method\": \"Affinity pulldown from brain extract, Reelin stimulation of cortical neurons, Rap1-GTP assay, site-specific phosphomutant analysis\",\n      \"journal\": \"Current biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — biochemical binding mapped to specific pY sites, functional GTPase assay, primary neurons, site-specific mutants required for neuron positioning in vivo\",\n      \"pmids\": [\"15062102\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Compound heterozygosity of Crkl and Tbx1 (mouse homologs of 22q11 genes) drastically increases penetrance of DiGeorge syndrome-like phenotype; Crkl and Tbx1 have dose-dependent functions in pharyngeal segmentation and RA signaling; partial rescue is achieved by genetically reducing RA levels, indicating that del22q11 involves dose-sensitive interaction of CRKL and TBX1 via locally aberrant RA signaling.\",\n      \"method\": \"Compound heterozygous mouse genetics, epistasis with RA synthesis gene, embryo phenotype analysis\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis in vivo with partial rescue, establishing pathway position and interaction\",\n      \"pmids\": [\"16399080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Crkl is required for normal cellular responses to Fgf8 (survival, migration, Erk activation, target gene expression); Fgf8 induces tyrosine phosphorylation of FgfR1 and FgfR2 and their direct binding to Crkl; genetic interactions exist between Crkl and Fgf8 in pharyngeal/cardiac morphogenesis.\",\n      \"method\": \"Crkl-/- mouse embryos, Fgf8 stimulation assay, receptor phosphorylation, Erk activation assay, genetic compound heterozygosity\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function in null cells/embryos with receptor binding and signaling readouts, epistasis with Fgf8\",\n      \"pmids\": [\"16399079\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"NS1 proteins of 1918 pandemic and avian H7N3 influenza viruses contain a consensus SH3-binding motif that binds specifically to the N-terminal SH3 domain of CrkL (and Crk) but not to other SH3 domains tested; endogenous CrkL co-precipitates NS1 from H7N3-infected cells; SH3 binding was associated with enhanced PI3K/Akt signaling.\",\n      \"method\": \"SH3 phage library screening, recombinant protein binding assay, co-immunoprecipitation from infected cells, Akt phosphorylation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phage library + recombinant binding + co-IP from infected cells with functional signaling readout, single lab\",\n      \"pmids\": [\"18165234\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Reducing expression of Crk and CrkL (but not BDNF pathway) blocks Reelin-stimulated dendritogenesis in hippocampal neurons (approximately twofold enhancement by Reelin); CrkL acts downstream of Dab1 phosphorylation in postnatal hippocampal dendrite development.\",\n      \"method\": \"Retroviral shRNA knockdown, hippocampal neuron culture, Reelin stimulation, dendrite morphometry\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA knockdown with specific Reelin pathway readout vs. BDNF control, single lab\",\n      \"pmids\": [\"18477607\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The WAVE2 complex recruits Abl kinase upon TCR ligation; WAVE2 regulates TCR-mediated Rap1 activation via membrane recruitment and activation of the CrkL–C3G exchange complex; Abl phosphorylates C3G (required for its GEF activity toward Rap1) but does not regulate CrkL–C3G membrane recruitment; this signaling node regulates integrin clustering and affinity maturation.\",\n      \"method\": \"Co-immunoprecipitation, Rap1-GTP assay, dominant-negative constructs, membrane fractionation, integrin activation assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods dissecting pathway with specific functional readouts in T cells, single lab\",\n      \"pmids\": [\"18809728\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"MUC1 ligation of ICAM-1 induces rapid formation of a Src–CrkL signaling complex at the MUC1 cytoplasmic domain; Src recruits CrkL to MUC1 and is required for CrkL-dependent Rac1- and Cdc42-mediated actin cytoskeletal protrusions at cell–cell contact sites.\",\n      \"method\": \"Co-immunoprecipitation, Src kinase inhibition, Rac1/Cdc42 GTPase assay, actin reorganization assay\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP with functional actin/GTPase assay, single lab\",\n      \"pmids\": [\"18403635\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"CRKL amplification and overexpression in NSCLC activates SOS1-RAS-RAF-ERK and SRC-C3G-RAP1 pathways; CRKL overexpression in EGFR-mutant cells induces resistance to gefitinib by activating ERK and AKT signaling; CRKL knockdown in CRKL-amplified NSCLC induces cell death.\",\n      \"method\": \"shRNA knockdown, overexpression in airway epithelial cells, anchorage-independent growth assay, pathway inhibitor studies\",\n      \"journal\": \"Cancer discovery\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — gain- and loss-of-function with pathway analysis, replicated in multiple NSCLC cell lines\",\n      \"pmids\": [\"22586683\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"PI3K p85 subunit interacts with CrkL; this p85–CrkL interaction is required for NKG2D-mediated NK cell cytotoxicity; CrkL depletion impairs NK–target conjugate formation, MTOC polarization, and granule release; Rap1 is activated downstream of NKG2D in a PI3K- and CrkL-dependent manner.\",\n      \"method\": \"PI3K inhibitor treatment, siRNA depletion of CrkL, conjugate assay, MTOC polarization assay, Rap1-GTP assay\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockdown with multiple functional readouts and signaling pathway dissection, single lab\",\n      \"pmids\": [\"19454690\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Molecular dynamics modeling and in vitro binding studies show that pY463 in FGFR1 binds specifically to the CRKL SH2 domain with ~30-fold higher affinity than to CRK SH2; CRKL (but not CRK) is an essential component of an FGF8-induced feed-forward loop enabling efficient Erk1/2 activation and anchorage-independent growth.\",\n      \"method\": \"Molecular dynamics simulation, in vitro binding assay with phosphopeptide, Crkl-null cells, Erk activation assay, anchorage-independent growth assay\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — structural modeling validated by in vitro binding affinity measurement, loss-of-function in null cells with functional readouts, single lab\",\n      \"pmids\": [\"19307307\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Agrin stimulates phosphorylation of two tyrosine residues in the C-terminal domain of Dok-7, leading to recruitment of CrkL and Crk; selective inactivation of Crk and CrkL in skeletal muscle causes severe neuromuscular synapse defects in vivo, placing CrkL downstream of MuSK–Dok-7 in presynaptic and postsynaptic differentiation.\",\n      \"method\": \"In vivo muscle-specific knockout, tyrosine phosphorylation mapping of Dok-7, co-IP\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — in vivo tissue-specific double knockout with clear NMJ phenotype, mapped to specific phosphotyrosine sites on Dok-7\",\n      \"pmids\": [\"21041412\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"CRKL is specifically required for p210BCR-ABL-induced IL-3-independent growth of myeloid progenitors and B-lymphoid cell outgrowth; a synthetic phosphotyrosyl peptide binding the CRKL SH2 domain blocks CRKL association with the BCR-ABL complex and reduces c-MYC levels in leukemic cells.\",\n      \"method\": \"CrkL-null hematopoietic progenitor cells, fetal liver transplantation, SH2-blocking peptide, immunoprecipitation\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — loss-of-function in null progenitor cells plus pharmacological SH2-domain blocking with defined molecular readout, single lab\",\n      \"pmids\": [\"20807813\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"NMR and structural analysis reveals CRKL and phospho-CRKL have markedly different structures from CrkII and phospho-CrkII; the SH2 and SH3 domain binding activities are regulated differently in the two proteins; CRKL forms a constitutive complex with Abl (unlike CrkII), explaining the strong preference of Bcr-Abl for CRKL over CrkII.\",\n      \"method\": \"NMR structural analysis, binding activity assays\",\n      \"journal\": \"Nature chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structural determination with functional binding validation, explains mechanistic preference for CRKL, single lab\",\n      \"pmids\": [\"22581121\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"In rhabdomyosarcoma, CRKL signaling is associated with YES kinase (a Src family kinase); CRKL is required for RMS tumor growth in vitro and in vivo; no interaction of CRKL with IGFIR, MET, or PI3K/AKT/mTOR pathways was detected in this context.\",\n      \"method\": \"shRNA loss-of-function screen, co-immunoprecipitation, in vivo xenograft assay, kinase pathway inhibitors\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — unbiased shRNA screen validated with in vivo assay and signaling pathway dissection, single lab\",\n      \"pmids\": [\"23318429\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Loss of both Crk and CrkL in fibroblasts (conditional double knockout) causes rounded morphology, loss of focal adhesions, reduced actin stress fibers, microtubule collapse, decreased motility and wound healing, reduced p130Cas phosphorylation; reintroduction of CrkII partially rescues the phenotype; individual deletion of Crk or CrkL alone produces modest phenotype, indicating overlapping but essential roles.\",\n      \"method\": \"Conditional Crk/CrkL double knockout fibroblasts, morphology and motility assays, immunofluorescence, actin stabilization rescue\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic conditional double knockout with multiple orthogonal readouts and genetic rescue, single lab\",\n      \"pmids\": [\"24166500\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CrkL overexpression enhances lung cancer cell invasion through ERK-MMP9 pathway; CRKL activates AP-1 (c-fos binding to MMP9 promoter) via ERK phosphorylation; ERK inhibitor PD98059 blocks CRKL-induced invasion and MMP9 expression.\",\n      \"method\": \"Matrigel invasion assay, chromatin immunoprecipitation, luciferase reporter, ERK inhibitor treatment, Western blotting\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional invasion assay with pathway inhibition and promoter occupancy, single lab\",\n      \"pmids\": [\"24664993\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CrkL and CrkL form hetero-oligomers with Crk2; both are recruited to tyrosine-phosphorylated nephrin in podocytes; simultaneous deletion of Crk1/2 and CrkL in podocytes causes albuminuria and altered foot process architecture; CrkL is required for nephrin-induced lamellipodia formation in podocytes in vitro.\",\n      \"method\": \"Podocyte-specific double conditional knockout mice, co-immunoprecipitation, in vitro lamellipodia assay\",\n      \"journal\": \"Kidney international\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — tissue-specific double knockout with molecular rescue and in vitro mechanistic assay, single lab\",\n      \"pmids\": [\"24499776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CrkL is required for SDF-1-induced activation of the Raf-1/MEK/Erk pathway through Ras and Rac in hematopoietic cells; CrkL overexpression enhances SDF-1-induced chemotaxis; Lyn mediates SDF-1-induced CrkL tyrosine phosphorylation; the N-terminal SH3 domain (GEF-binding) is required for CrkL-enhanced Erk activation and chemotaxis.\",\n      \"method\": \"Overexpression, CrkL N-SH3 deletion mutant, Ras/Rac GTP assay, MEK inhibitor, dominant-negative GTPases, Src inhibitor PP1\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple pathway inhibitors with domain deletion mutants and functional chemotaxis assay, single lab\",\n      \"pmids\": [\"16781119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CRKL is dosage-sensitive for cardiac outflow tract development; haploinsufficiency of CRKL can cause conotruncal heart defects in humans with atypical distal 22q11.2 deletions that exclude TBX1; a Crkl hypomorphic allele series in mice shows spectrum of heart defects dependent on Crkl expression level.\",\n      \"method\": \"Crkl allelic series (null + hypomorphic), mouse cardiac phenotyping, human genomic deletion mapping\",\n      \"journal\": \"American journal of human genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — allelic series with dose-dependent phenotype in mice correlated with human genetic evidence\",\n      \"pmids\": [\"25658046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Sorbs1 and Sorbs2 associate with CrkL (identified by mass spectrometry of CrkL-binding proteins); Sorbs1/2 are required for AChR clustering in vitro and are localized at neuromuscular synapses in vivo, acting as redundant proteins downstream of the MuSK/Dok-7/Crk/CrkL complex.\",\n      \"method\": \"Mass spectrometry pulldown, CrkL co-immunoprecipitation, AChR clustering assay, in vivo NMJ immunostaining\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — MS-based interaction partner identification with functional validation in vitro and in vivo, single lab\",\n      \"pmids\": [\"26527617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CRKL is a direct transcriptional target de-repressed by Zeb1-dependent EMT; CRKL serves as a downstream effector of miR-200 loss that mediates focal adhesion formation and outside-in signaling through integrin β1 (ECM→FAK/Src→CRKL) as well as inside-out signaling maintaining tumor cell–matrix contacts; CRKL knockdown suppresses experimental metastases in vivo without affecting primary tumor growth.\",\n      \"method\": \"miR-200 gain/loss-of-function, CRKL siRNA knockdown, 3D invasion assay, in vivo experimental metastasis model, focal adhesion immunofluorescence\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional loss-of-function with in vivo metastasis readout and mechanistic signaling assays, single lab\",\n      \"pmids\": [\"26728244\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CRKL interacts with tyrosine-phosphorylated p130Cas in PTEN-null cancer cells and is required for p110β-dependent PI3K signaling and cell proliferation in these cells; Src phosphorylates and activates p130Cas, linking PTEN loss to p110β activation via CRKL; Src and p110β inhibition cooperate to suppress PTEN-null cell growth.\",\n      \"method\": \"CRKL knockdown, co-immunoprecipitation with p130Cas, PI3K signaling assay, Src inhibitor epistasis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — knockdown with defined signaling pathway readout and epistasis, single lab\",\n      \"pmids\": [\"28723560\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"CRKL expression is dosage-sensitive for genitourinary development; Crkl null mice have upper GU defects (23% rate); Crkl heterozygous males have cryptorchidism, lower testis weight, reduced sperm count and subfertility; RNA-seq of Crkl mutant kidneys reveals 52 differentially regulated genes including a 5-fold upregulation of Foxd1 (a nephron progenitor regulator).\",\n      \"method\": \"Crkl null/heterozygous mice, microdissection phenotyping, RNA-sequencing of developing kidneys\",\n      \"journal\": \"Proceedings of the National Academy of Sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with transcriptomic pathway analysis in relevant tissue, multiple GU phenotypes documented\",\n      \"pmids\": [\"28439006\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"SASH1 interacts with CRKL (identified by yeast two-hybrid and co-IP/MS); SASH1 inhibits CRKL-mediated SRC kinase activation, which is required for EMT; SASH1-deficient cells form significantly more metastases in vivo in a manner entirely dependent on CRKL; CRKL overexpression is associated with poor chemotherapy response.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation/mass spectrometry, domain mapping, site-directed mutagenesis, CRISPR knockout, in vivo orthotopic mouse model\",\n      \"journal\": \"Cellular and molecular gastroenterology and hepatology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods for interaction, in vivo genetic epistasis, CRISPR KO, single lab\",\n      \"pmids\": [\"30480076\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CRKL inhibits APC-mediated proteasomal degradation of β-catenin by competitively decreasing Axin1 binding to APC, thereby promoting VEGFα and CXCL1 expression; this leads to tumor-associated neutrophil infiltration that blocks CD8+ T cell function and creates anti-PD-1 resistance in hepatocellular carcinoma.\",\n      \"method\": \"Pooled genetic screen, CRISPR-Cas9 knockout, transgenic mouse model (Alb-Cre/Trp53fl/fl + CRKL vector), mass cytometry, co-immunoprecipitation, patient-derived organotypic tumor spheroid model\",\n      \"journal\": \"Journal of hepatology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal in vitro and in vivo methods with mechanistic co-IP identifying β-catenin/Axin1 interaction, validated in patient models\",\n      \"pmids\": [\"38403027\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CRKL is an SH2-SH3-SH3 adaptor protein that functions as a major convergence point in tyrosine kinase signaling: its SH2 domain binds phosphotyrosine-containing proteins (pCBL, pCAS/HEF1, paxillin, p130Cas, pGAB1, pDab1, pDok-7, pFGFR1-pY463, pSTAT5, pEpoR), while its N-terminal SH3 domain constitutively binds guanine nucleotide exchange factors (C3G, SOS) and kinases (c-ABL, BCR/ABL), thereby coupling upstream receptor and oncogenic tyrosine kinase signals to Ras/ERK, Rap1, Rac1/Cdc42, and JNK pathways; BCR/ABL phosphorylates CRKL specifically at Y207 (which acts as a negative regulatory autoinhibitory site); in its unphosphorylated state CRKL forms a constitutive complex with Abl (explaining Bcr-Abl's preference for CRKL over CrkII as shown structurally by NMR); CRKL also functions as a nuclear adaptor that complexes with phospho-STAT5 to regulate transcription at GAS elements downstream of type I interferons and cytokines; in developmental contexts CRKL mediates FGF8 receptor signaling (via direct pFGFR1-Y463 binding) and is required downstream of Reelin/Dab1, Dok-7/MuSK, and Src/Cas for neural crest development, neuromuscular synapse formation, podocyte morphogenesis, genitourinary development, and integrin-induced cell migration; additionally, CRKL inhibits APC-mediated β-catenin degradation by displacing Axin1 to activate a VEGFα/CXCL1/tumor-associated neutrophil axis relevant to immune checkpoint resistance.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CRKL is an SH2–SH3–SH3 adaptor protein that couples tyrosine kinase signals to small-GTPase and Ras/ERK pathways, serving as a convergence point in oncogenic, cytokine, and developmental signaling [#0, #16]. Its modular architecture partitions binding labor: the SH2 domain docks onto phosphotyrosine-containing partners while the N-terminal SH3 domain constitutively engages downstream effectors. Through its SH2 domain CRKL binds tyrosine-phosphorylated CBL, paxillin, p130Cas/HEF1, GAB1, Dab1, Dok-7, the FGFR1 pY463 site, and STAT5 [#3, #4, #7, #23, #35, #44, #45], while its N-terminal SH3 domain constitutively binds the guanine-nucleotide exchange factors C3G and SOS and the kinases c-ABL/BCR-ABL [#2, #10, #11]. This dual engagement allows CRKL to relay receptor and oncogene input to Ras/ERK, Rap1, and Rac1/Cdc42 outputs, principally by membrane-recruiting C3G to activate Rap1 and integrins [#11, #21, #22] and by recruiting DOCK family GEFs at focal adhesions to activate Rac1/Cdc42 during integrin- and Src/p130Cas-driven migration [#30, #33]. In BCR-ABL leukemia CRKL is the major phosphorylated adaptor, forming a constitutive complex with Abl that explains Bcr-Abl's preference for CRKL over CrkII as resolved by NMR; phosphorylation at Y207 acts as a negative-regulatory autoinhibitory site, and CRKL is required for BCR-ABL-driven factor-independent growth [#1, #8, #16, #46, #47]. Beyond the cytoplasm, IFNα-activated Tyk-2/Jak-1 phosphorylate STAT5, which docks the CRKL SH2 domain; the CRKL–STAT5 complex translocates to the nucleus and binds GAS-element promoters, making CRKL a nuclear transcriptional adaptor [#13, #18, #32]. CRKL is dosage-sensitive in development: Crkl loss phenocopies DiGeorge syndrome, genetically interacts with Tbx1 through retinoic-acid signaling, and mediates FGF8/FGFR, Reelin/Dab1, and MuSK/Dok-7 signaling required for neural crest, cardiac outflow tract, neuromuscular synapse, podocyte, and genitourinary development [#26, #35, #36, #37, #45, #51, #53, #57]. In cancer, CRKL amplification activates SOS1-Ras-ERK and Src-C3G-Rap1 to drive NSCLC growth and EGFR-inhibitor resistance, promotes invasion and metastasis, and confers anti-PD-1 resistance in hepatocellular carcinoma by stabilizing β-catenin through displacement of Axin1 from APC [#42, #55, #58, #59].\",\n  \"teleology\": [\n    {\n      \"year\": 1993,\n      \"claim\": \"Established CRKL as a distinct adaptor-encoding gene, defining its SH2-SH3-SH3 architecture and predicting a signal-transduction role before any function was known.\",\n      \"evidence\": \"cDNA cloning and sequence/domain analysis of a 22q11 gene\",\n      \"pmids\": [\"8361759\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No functional validation beyond sequence\", \"No binding partners or pathway defined\"]\n    },\n    {\n      \"year\": 1994,\n      \"claim\": \"Identified CRKL as the principal tyrosine-phosphorylated adaptor in BCR-ABL leukemia and a direct ABL interactor, anchoring CRKL within oncogenic kinase signaling.\",\n      \"evidence\": \"Anti-phosphotyrosine immunoblotting of CML cells, yeast two-hybrid, co-IP, and in vitro kinase assays\",\n      \"pmids\": [\"8083188\", \"7524758\", \"7521685\", \"8168080\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphorylation site on CRKL not yet mapped\", \"Functional consequence of complex formation undefined\"]\n    },\n    {\n      \"year\": 1996,\n      \"claim\": \"Mapped CRKL's bipartite binding logic—SH2 to phospho-CBL/paxillin/p130Cas, SH3 to ABL/SOS—and showed CRKL transforms fibroblasts and activates Ras/JNK, defining it as a functional signaling node not a passive scaffold.\",\n      \"evidence\": \"GST-domain pulldowns, site-directed mutagenesis, co-IP, fibroblast transformation and Ras/JNK activation assays\",\n      \"pmids\": [\"7545163\", \"7493940\", \"8632906\", \"8798523\", \"8810278\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endogenous (non-overexpression) requirement not yet tested\", \"Domain contributions to transformation not fully dissected\"]\n    },\n    {\n      \"year\": 1997,\n      \"claim\": \"Defined Y207 as the major BCR/ABL phosphorylation site acting as an autoinhibitory negative regulator, and showed the N-SH3–C3G axis drives Rap1 activation via membrane recruitment, establishing the core regulatory and effector mechanisms.\",\n      \"evidence\": \"Phosphopeptide mapping, Y207F mutagenesis, C3G co-expression with Rap1 GTP-binding and farnesylation-rescue assays\",\n      \"pmids\": [\"9053848\", \"9268367\", \"9162067\", \"9092574\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of Y207 autoinhibition not resolved\", \"Physiological triggers regulating Y207 phosphorylation unclear\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Confirmed Y207 phosphorylation suppresses CRKL function while SH2 and N-SH3 domains are essential for activity, and identified a CrkL/Crk-selective SH3-binding peptide, providing tools and mechanistic clarity on domain requirements.\",\n      \"evidence\": \"Y207F mutagenesis with transformation/JNK/adhesion readouts and selective SH3-binding peptide complex-disruption assays\",\n      \"pmids\": [\"9710592\", \"9591773\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endogenous regulation of phospho-Y207 in normal cells unaddressed\", \"Peptide selectivity in vivo not established\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Revealed CRKL as a dual cytoplasmic-nuclear adaptor—coupling Epo/IL-3/integrin signals to Ras/ERK, Rap1, and adhesion via C3G, while also forming nuclear STAT5–DNA complexes at GAS-like elements to regulate IFN-stimulated transcription.\",\n      \"evidence\": \"Migration/adhesion assays with domain mutants, dominant-negative C3G, ERK/Ras assays, EMSA, nuclear fractionation, HPK1-JNK assays\",\n      \"pmids\": [\"10608804\", \"10339478\", \"10514505\", \"9872990\", \"9891069\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of CRKL nuclear import unknown\", \"Direct vs indirect DNA contact by CRKL not resolved\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Demonstrated CRKL activates Rac1 through DOCK GEFs and is genetically required for IFNα-dependent GAS transcription and Rap1 activation, extending the effector repertoire to actin-regulatory GTPases and confirming nuclear function in loss-of-function cells.\",\n      \"evidence\": \"DOCK2 co-IP and Rac1-GTP assays, CrkL-null MEF reporter and Rap1 assays with dominant-negative kinases\",\n      \"pmids\": [\"12393632\", \"11866427\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Selectivity among DOCK family GEFs unclear\", \"How one adaptor coordinates Rap1 vs Rac1 outputs spatially undefined\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Showed Src-dependent, p130Cas-mediated recruitment of CRKL to focal adhesions activates Rac1/Cdc42 and is required for integrin-induced migration even when CrkII is present, establishing a non-redundant CRKL function in adhesion signaling.\",\n      \"evidence\": \"CrkL-null MEF rescue, forced-localization chimeras, GTPase and haptotaxis assays, ASAP1 pulldown/MS\",\n      \"pmids\": [\"12665586\", \"12522101\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Basis for CrkL/CrkII functional non-equivalence at this stage unexplained\", \"In vivo relevance of FA recruitment not yet shown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Established CRKL as a dosage-sensitive 22q11 gene whose loss phenocopies DiGeorge syndrome, acting in FGF8/FGFR and Tbx1/retinoic-acid pathways during neural crest and pharyngeal development.\",\n      \"evidence\": \"Crkl knockout and compound-heterozygous mouse genetics, Fgf8 stimulation/receptor binding, ERK assays, RA-pathway epistasis\",\n      \"pmids\": [\"11242111\", \"16399080\", \"16399079\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct molecular link between CRKL and RA signaling unresolved\", \"Cell-autonomous vs non-autonomous neural crest roles not separated\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified CRKL amplification as a lung cancer driver activating SOS1-Ras-ERK and Src-C3G-Rap1 and conferring EGFR-inhibitor resistance, and resolved the FGFR1 pY463 SH2 interaction structurally to explain CRKL's ~30-fold selectivity over CRK.\",\n      \"evidence\": \"shRNA/overexpression in NSCLC with pathway inhibitors, molecular dynamics and phosphopeptide binding affinity, Crkl-null ERK/growth assays\",\n      \"pmids\": [\"22586683\", \"19307307\", \"19454690\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Generality of CRKL amplification across tumor types unquantified\", \"Structural model not validated by crystallography\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Resolved by NMR why Bcr-Abl prefers CRKL over CrkII—CRKL forms a constitutive Abl complex with distinct domain regulation—providing the structural basis for CRKL's central role in CML.\",\n      \"evidence\": \"NMR structural analysis of CRKL and phospho-CRKL with binding-activity assays\",\n      \"pmids\": [\"22581121\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full-length CRKL conformational dynamics not captured\", \"Structural state in cells inferred, not directly observed\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Established CRKL as functionally required for BCR-ABL leukemogenesis and as a tractable SH2-domain target, linking adaptor function to c-MYC and transformed growth.\",\n      \"evidence\": \"CrkL-null progenitor transplantation, SH2-blocking peptide, immunoprecipitation\",\n      \"pmids\": [\"20807813\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Therapeutic window of SH2 blockade untested\", \"Downstream effectors beyond c-MYC not enumerated\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Placed CRKL downstream of Reelin/Dab1 and MuSK/Dok-7 in vivo, establishing essential, partly redundant Crk/CrkL roles in cortical neuron positioning, dendritogenesis, and neuromuscular synapse formation.\",\n      \"evidence\": \"Tissue-specific Crk/CrkL knockouts, Dab1/Dok-7 phosphotyrosine mapping, Rap1 assays, NMJ and dendrite phenotyping\",\n      \"pmids\": [\"15062102\", \"18477607\", \"21041412\", \"18809728\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional distinction between Crk and CrkL at synapses unresolved\", \"Direct effectors at the NMJ not fully mapped\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated broad essential adaptor function via Crk/CrkL double knockout—loss of focal adhesions, stress fibers, and motility—and tissue-specific roles in podocyte and genitourinary development, showing overlapping but indispensable cytoskeletal regulation.\",\n      \"evidence\": \"Conditional double-knockout fibroblasts and podocytes, morphology/motility/lamellipodia assays, Crkl allelic series and RNA-seq\",\n      \"pmids\": [\"24166500\", \"24499776\", \"25658046\", \"26527617\", \"28439006\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular reason single deletions are mild not fully explained\", \"Direct transcriptional targets in GU development (e.g., Foxd1 regulation) mechanism unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined CRKL as a metastasis and EMT effector regulated by Zeb1/miR-200 and antagonized by SASH1, linking CRKL-driven Src/p130Cas-PI3K signaling to invasion and poor therapy response.\",\n      \"evidence\": \"miR-200 modulation, CRKL knockdown, in vivo metastasis models, SASH1 yeast two-hybrid/co-IP/MS, CRISPR epistasis, PTEN-null PI3K assays\",\n      \"pmids\": [\"26728244\", \"30480076\", \"28723560\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CRKL's adaptor or scaffold activity drives EMT is unresolved\", \"Context-dependence across tumor types not systematically tested\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Uncovered a non-canonical CRKL function in immune evasion—stabilizing β-catenin by displacing Axin1 from APC to drive a VEGFα/CXCL1/neutrophil axis that confers anti-PD-1 resistance—extending CRKL biology beyond tyrosine-kinase adaptation.\",\n      \"evidence\": \"Pooled CRISPR screen, transgenic HCC mouse model, mass cytometry, co-IP, patient-derived tumor spheroids\",\n      \"pmids\": [\"38403027\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of CRKL–APC/Axin1 competition unresolved\", \"How tyrosine-adaptor functions relate to this β-catenin role unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How CRKL's spatial and temporal switching among Rap1, Rac1/Cdc42, ERK, nuclear STAT5, and β-catenin outputs is selected in a given cell context remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No unified model linking domain occupancy to pathway choice\", \"Mechanism of nuclear import undefined\", \"Endogenous regulation of Y207 phosphorylation in normal physiology unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 10, 11, 18, 33, 49]},\n      {\"term_id\": \"GO:0005515\", \"supporting_discovery_ids\": [3, 4, 7, 23, 35, 44]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [10, 33]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [18, 24, 32]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [30, 33, 49]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [11, 22, 33, 42]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [26, 36, 37, 45, 53, 57]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [1, 42, 58, 59]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [32, 40, 43]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [18, 24, 32]}\n    ],\n    \"complexes\": [\n      \"BCR/ABL–CRKL–CBL complex\",\n      \"CRKL–STAT5 nuclear complex\",\n      \"CrkL–C3G complex\"\n    ],\n    \"partners\": [\n      \"ABL1\",\n      \"BCR\",\n      \"CBL\",\n      \"RAPGEF1\",\n      \"BCAR1\",\n      \"STAT5\",\n      \"DOCK2\",\n      \"PIK3R1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":7,"faith_total":7,"faith_pct":100.0}}