{"gene":"DLC1","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2003,"finding":"DLC1 protein functions as a GTPase-activating protein (GAP) specific for RhoA and Cdc42, as established by in vitro GTPase activating protein activity assay.","method":"In vitro GAP activity assay","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 1 / Strong — direct in vitro enzymatic assay, replicated across multiple labs in subsequent studies","pmids":["14633684"],"is_preprint":false},{"year":2008,"finding":"Full-length DLC1 exhibits strong GAP activity for RhoA, RhoB, and RhoC but only very limited activity for Cdc42 in vitro; in contrast, the isolated RhoGAP domain showed 5- to 20-fold enhanced activity for RhoA, RhoB, RhoC, and Cdc42, suggesting the flanking domains impose autoinhibition. DLC1 reduces RhoA activity at the leading edge of cellular protrusions, and its anti-tumor activity involves both RhoGAP-dependent and RhoGAP-independent mechanisms. Unlike rat p122RhoGAP, human DLC1 was not capable of activating phospholipase C-delta1.","method":"In vitro GAP activity assay, RhoA biosensor live imaging, cell-based assays","journal":"Molecular carcinogenesis","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro biochemical assay plus live-cell biosensor imaging, multiple orthogonal methods in single study","pmids":["17932950"],"is_preprint":false},{"year":2008,"finding":"DLC1 negatively regulates the Rho/ROCK/MLC2 pathway in hepatocellular carcinoma. Ectopic DLC1 expression abrogates Rho/ROCK-mediated cytoskeletal reorganization (stress fibers, focal adhesions) and downregulates cortical phosphorylation of myosin light chain 2 (MLC2) and MYPT1 phosphorylation at Thr853. A RhoGAP-deficient mutant (K714E) abolishes these inhibitory effects, confirming RhoGAP-dependency. Dominant-active ROCK rescues cells from DLC1-induced cytoskeletal collapse.","method":"Immunofluorescence, western blot, dominant-active ROCK epistasis, RhoGAP-dead mutant","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (immunofluorescence, western blot, mutant rescue, epistasis), single lab","pmids":["18648664"],"is_preprint":false},{"year":2008,"finding":"The SAM domain of DLC1 functions as an autoinhibitory domain of intrinsic RhoGAP activity. The SAM and START domains are dispensable for DLC1 association with focal adhesions. A dominant-negative N-terminal fragment displaces endogenous DLC1 from focal adhesions and profoundly inhibits cell migration.","method":"Structure-function analysis with truncation and missense mutants, cell morphology and migration assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — systematic domain mutagenesis combined with functional assays, multiple orthogonal readouts","pmids":["18786931"],"is_preprint":false},{"year":2008,"finding":"DLC1 interacts with 14-3-3 adaptor proteins via phosphoserine recognition motifs involving Ser327 and Ser431. Phorbol-ester-induced activation of PKC/PKD stimulates this association. Binding to 14-3-3 inhibits DLC1 GAP activity, facilitates RhoA signaling, and blocks DLC1 nucleocytoplasmic shuttling by masking a nuclear localization sequence.","method":"Co-immunoprecipitation, in vitro GAP assay, site-directed mutagenesis, subcellular fractionation/imaging","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — co-IP, in vitro GAP activity, mutagenesis of binding motifs, multiple orthogonal methods","pmids":["19066281"],"is_preprint":false},{"year":2009,"finding":"p120Ras-GAP (RASA1) interacts with DLC1 and colocalizes in focal adhesions. The Ras-GAP SH3 domain binds the DLC1 RhoGAP domain and inhibits DLC1 RhoGAP activity in vitro. Overexpression of Ras-GAP impairs DLC1 growth-suppressing activity and increases RhoA activity in vivo.","method":"Co-immunoprecipitation, in vitro GAP assay with purified proteins, cell growth assay, active RhoA pulldown","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro reconstitution with purified proteins plus cellular functional assays, multiple orthogonal methods","pmids":["19151751"],"is_preprint":false},{"year":2009,"finding":"The SAM domain of DLC1 binds eukaryotic elongation factor 1A1 (EF1A1) but not the SAM domain of DLC2. The solution structure of DLC1 SAM reveals a monomeric fold with four parallel helices. A hydrophobic patch (F38, L39, F40) is required for EF1A1 interaction. SAM-EF1A1 interaction facilitates EF1A1 distribution to membrane periphery upon growth factor stimulation and contributes to DLC1-mediated suppression of cell migration independent of the RhoGAP domain.","method":"Protein precipitation, mass spectrometry, NMR structure, mutagenesis, cell migration assay","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure plus mutagenesis plus functional assays, multiple orthogonal methods in single study","pmids":["19158340"],"is_preprint":false},{"year":2009,"finding":"DLC1 activation requires binding to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] through a polybasic region (PBR) adjacent to the RhoGAP domain. PI(4,5)P2-containing membranes stimulate DLC1 GAP activity in vitro. A PBR-deficient DLC1 mutant is severely compromised in suppressing cell spreading, directed migration, and proliferation.","method":"Lipid-binding assay, in vitro GAP activity assay with PI(4,5)P2 liposomes, mutagenesis, cell-based assays","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with lipids, GAP activity assay, mutagenesis, multiple functional readouts","pmids":["19710422"],"is_preprint":false},{"year":2009,"finding":"Tensin1 requires its SH2 domain (R1488) to bind DLC1; tensin1 F302A mutation (abrogating PP1alpha binding) also reduces DLC1 association, linking PP1alpha and DLC1 to tensin1 signaling. DLC1 binding via tensin mediates Rho regulation, but PP1alpha has additional DLC1-independent effects on migration and invasion.","method":"Co-immunoprecipitation, mutagenesis, cell migration and invasion assays, MLC20 phosphorylation western blot","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with mutagenesis, multiple functional readouts, single lab","pmids":["19826001"],"is_preprint":false},{"year":2009,"finding":"Tensin2 binds DLC1 via its PTB domain at a novel binding site (residues 375–385 of DLC1). Deletion of this PTB-binding site partially reduces DLC1 RhoGAP activity and attenuates growth-suppressive activity without affecting focal adhesion localization or tensin1/cten interactions.","method":"Co-immunoprecipitation, deletion mutagenesis, RhoGAP activity assay, growth suppression assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with deletion mutants, RhoGAP activity assay, growth assay, single lab","pmids":["19440389"],"is_preprint":false},{"year":2010,"finding":"Tensin2 knockdown significantly reduces the ability of human foreskin fibroblasts to contract 3D collagen gels, associated with reduced Rho activity; this inhibition is reversed by depletion of DLC1, placing DLC1 downstream of tensin2 in regulating Rho-mediated actomyosin contraction and collagen remodeling.","method":"siRNA knockdown, 3D collagen gel contraction assay, RhoA activity pulldown, epistasis by double knockdown","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis by double knockdown with defined functional readout, single lab","pmids":["20069572"],"is_preprint":false},{"year":2011,"finding":"DLC1 binds S100A10 (p11) in the cell cytoplasm; the interaction is mediated by central sequences in DLC1 and the C-terminus of S100A10. DLC1 competes with Annexin 2 for S100A10 binding, displacing S100A10 from Annexin 2 and making it accessible to ubiquitin-dependent degradation, thereby decreasing S100A10 steady-state levels and attenuating plasminogen activation. This mechanism inhibits cell migration, invasion, and anchorage-independent growth independently of DLC1 RhoGAP activity.","method":"Co-immunoprecipitation, colocalization imaging, competition assay, RhoGAP activity assay, cell invasion/migration assays","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, multiple orthogonal functional assays, competition biochemistry, single lab","pmids":["21372205"],"is_preprint":false},{"year":2011,"finding":"DLC1 contains an 8-aa LD-like motif (residues 469LDDILYHV476) that is necessary for binding to talin and FAK. This motif is required for DLC1 localization to focal adhesions and for full tumor suppressor activity. FAK binding is independent of talin and vice versa. Mutants deficient in talin/FAK binding have impaired tumor suppressor activity despite retained ability to negatively regulate overall Rho-GTP.","method":"Co-immunoprecipitation, mutagenesis, focal adhesion localization imaging, bioassays (growth inhibition, colony formation)","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — co-IP, mutagenesis, localization imaging, multiple functional assays, single lab with multiple orthogonal methods","pmids":["21969587"],"is_preprint":false},{"year":2012,"finding":"DLC1 forms a complex with α-catenin; binding is mediated by DLC1 N-terminal residues 340–435 and α-catenin residues 117–161. DLC1-α-catenin colocalize in cytosol and plasma membrane, where they associate with E-cadherin and β-catenin. DLC1-α-catenin complex reduces RhoA-GTP at the plasma membrane, increases E-cadherin mobility, and stabilizes adherens junctions. DLC1 GAP activity is required for α-catenin accumulation at the plasma membrane.","method":"Co-immunoprecipitation, mutagenesis, colocalization immunofluorescence, active Rho pulldown, E-cadherin FRAP, cell-based functional assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with domain mapping, multiple orthogonal methods, single lab","pmids":["22473989"],"is_preprint":false},{"year":2012,"finding":"Tensin3 activates DLC1 RhoGAP activity by binding DLC1 through its actin-binding domain, thereby releasing an autoinhibitory interaction between the SAM and RhoGAP domains of DLC1. Cten (C-terminal tensin-like protein), which lacks the actin-binding domain, does not activate DLC1. Tensin3 depletion augments actin stress fibers and focal adhesions and enhances cell motility via RhoA/ROCK signaling.","method":"Co-immunoprecipitation, in vitro RhoGAP activity assay, siRNA knockdown, actin/focal adhesion imaging, ROCK inhibitor rescue","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro RhoGAP activity assay plus cellular epistasis with multiple orthogonal methods","pmids":["22307599"],"is_preprint":false},{"year":2012,"finding":"DLC1 forms a complex with caveolin-1 (CAV-1); the interaction is mapped to the DLC1 START domain. Mutation of the START domain disrupts interaction and colocalization with CAV-1 and abolishes suppression of neoplastic growth, even though RhoA regulation is retained. This defines a RhoGAP-independent tumor suppressor mechanism mediated by the START domain-CAV-1 interaction.","method":"Co-immunoprecipitation, domain mutagenesis, colocalization imaging, anchorage-independent growth assay, active RhoA pulldown","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, domain mutagenesis, multiple functional assays, single lab","pmids":["22693251"],"is_preprint":false},{"year":2013,"finding":"DLC1 is ubiquitinated and degraded by the CRL4A (cullin 4A-RING ubiquitin ligase) complex via DDB1 and FBXW5 substrate receptor, providing a post-translational mechanism for DLC1 loss in NSCLC. siRNA suppression of cullin 4A, DDB1, or FBXW5 restores DLC1 protein, reduces RhoA-GTP, and causes DLC1-dependent decreases in NSCLC proliferation.","method":"siRNA knockdown, co-immunoprecipitation (ubiquitin ligase complex), ubiquitination assay, active RhoA pulldown, cell proliferation assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — co-IP of ubiquitin ligase complex, siRNA epistasis, multiple functional readouts, single lab","pmids":["24082123"],"is_preprint":false},{"year":2013,"finding":"PKA phosphorylates DLC1 at Ser549, enhancing its RhoGAP activity and promoting its tumor suppressor functions (suppression of hepatoma cell growth, motility, and metastasis). Ser549 phosphorylation induces dimerization of DLC1, and inducible dimerization alone can rescue tumor suppressive and RhoGAP activities of a Ser549-deletion DLC1 mutant.","method":"Site-directed mutagenesis, kinase assay, co-immunoprecipitation (dimerization), RhoGAP activity assay, in vitro and in vivo tumor suppression assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — kinase assay, mutagenesis, dimerization co-IP, in vitro RhoGAP activity, multiple functional assays","pmids":["23511482"],"is_preprint":false},{"year":2013,"finding":"DLC1 expression in metastatic prostate carcinoma cells induces E-cadherin expression at the mRNA level through suppression of RhoA-GTP and RhoC-GTP via its RhoGAP function. Rho/ROCK inhibitors mimic this effect. Knockdown of RhoC (more than RhoA) increases E-cadherin. Constitutively active RhoA(V14) and RhoC(V14) could not be reversed by DLC1, confirming epistasis.","method":"Gene expression (RT-PCR/western blot), ROCK inhibitors, shRNA, constitutively active Rho mutants (epistasis), cell invasion assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with constitutively active mutants, RhoGAP-dependent mechanism validated, multiple orthogonal methods","pmids":["23376848"],"is_preprint":false},{"year":2014,"finding":"CDK5 is a major regulator of DLC1. CDK5 phosphorylates four serines in DLC1 N-terminal to the RhoGAP domain. When unphosphorylated, this N-terminal region acts as an autoinhibitory domain that binds the RhoGAP domain and places DLC1 in a closed, inactive conformation. CDK5 phosphorylation reduces this autoinhibitory binding and coordinately activates DLC1 localization to focal adhesions, RhoGAP activity, and ability to bind tensin and talin.","method":"Kinase assay (CDK5 phosphorylation), mutagenesis, co-immunoprecipitation (autoinhibitory domain interaction), focal adhesion localization imaging, RhoGAP activity assay, tensin/talin binding assay","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro kinase assay, co-IP autoinhibition, mutagenesis, multiple functional readouts, single lab with multiple orthogonal methods","pmids":["25452387"],"is_preprint":false},{"year":2014,"finding":"Loss of DLC1 activates Rho-ROCK signaling, which mediates SMAD3 linker region phosphorylation and TGF-β-induced expression of parathyroid hormone-like hormone (PTHLH), leading to osteoclast maturation and osteolytic bone metastasis. Pharmacological inhibition of Rho-ROCK reduces PTHLH production and breast cancer bone metastasis in vitro and in vivo.","method":"siRNA knockdown, ROCK inhibitor, phosphorylation analysis, PTHLH expression assay, in vivo mouse metastasis model","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (knockdown, inhibitor, in vivo model), mechanistic pathway defined, single lab","pmids":["24590291"],"is_preprint":false},{"year":2016,"finding":"The crystal structure of the talin R8 domain bound to the DLC1 LD motif reveals that the DLC1 LD helix binds the four-helix bundle of talin R8 in a canonical triple-helix arrangement. The same R8 surface interacts with paxillin LD1 and LD2 motifs. Key charged residues stabilize R8-LD interactions, and mutations at this interface disrupt DLC1 binding and function in cells.","method":"X-ray crystallography, mutagenesis, in vitro binding assay, cell-based functional assays","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure plus mutagenesis validated in vitro and in cells","pmids":["27265849"],"is_preprint":false},{"year":2017,"finding":"Multiple RTK ligands increase RhoA-GTP in cells via AKT activation. AKT phosphorylates three serines (S298, S329, S567) in DLC1 N-terminal to the RhoGAP domain, inducing strong binding of that N-terminal region to the RhoGAP domain and converting DLC1 from an open, active dimer to a closed, inactive monomer. This reduces RhoA-GTP hydrolysis, binding of other DLC1 ligands, colocalization with focal adhesions, and tumor suppressor activity. AKT inhibition has potent antitumor activity specifically in DLC1-positive cancers.","method":"Kinase assay, mutagenesis, co-immunoprecipitation, active RhoA pulldown, focal adhesion localization imaging, in vivo tumor model, isogenic DLC1+/- cell lines","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro kinase assay, mutagenesis, co-IP mechanism validation, multiple cell and in vivo models, multiple orthogonal methods","pmids":["29114068"],"is_preprint":false},{"year":2018,"finding":"Talin R8 domain unfolding by mechanical force is required for DLC1 downstream signaling. Using a talin mutant resistant to force-induced R8 unfolding, DLC1 signaling (RhoA regulation) and cell mechanics are dependent on talin unfolding status, defining a mechanotransduction mechanism where talin acts as a force-sensitive scaffold for DLC1.","method":"Protein engineering (force-resistant talin mutant), atomic force microscopy, traction force microscopy, biophysical assays","journal":"PLoS biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — protein engineering with biophysical validation (AFM), functional consequences measured, multiple orthogonal methods","pmids":["30028837"],"is_preprint":false},{"year":2018,"finding":"DLC1 depletion in endothelial cells on stiff substrates reduces cell stiffness and impairs ICAM-1 adhesome formation by preventing recruitment of filamin B, α-actinin-4, and cortactin to clustered ICAM-1, thereby impairing leukocyte spreading. DLC1 overexpression rescues ICAM-1 adhesome stabilization. This function is independent of the DLC1 GAP domain.","method":"siRNA knockdown, DLC1 overexpression rescue, ICAM-1 clustering assay, co-immunoprecipitation, stiffness measurement (AFM), leukocyte adhesion assay","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA knockdown with rescue, functional readout, GAP-independence shown, single lab","pmids":["30231995"],"is_preprint":false},{"year":2019,"finding":"The DLC1 SAM domain binds to specific peptide motifs within the C2 domains of tensin3 and PTEN. This SAM-C2 interaction mediates DLC1 activation. Peptides containing the C2 binding motifs block the C2-SAM interaction, promote DLC1 RhoGAP activity, decrease RhoA activation, and reduce tumor cell growth and migration. A cyclic version of the TNS3 C2-derived peptide enters cancer cells and effectively inhibits migration.","method":"Co-immunoprecipitation, in vitro RhoGAP activity assay, cell-penetrating peptide functional assays, soft agar growth, migration assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro biochemical reconstitution, RhoGAP activity assay, multiple functional cellular assays, single lab","pmids":["31806702"],"is_preprint":false},{"year":2020,"finding":"DLC1 is a direct transcriptional target of the activated YAP/TAZ-TEAD complex. Substrate stiffening and VEGF stimulate DLC1 expression in endothelial cells in a YAP/TAZ-dependent manner. DLC1 limits F-actin fiber formation, integrin-based focal adhesion lifetime, and traction forces. DLC1 depletion impairs endothelial cell polarization in directed collective migration and inhibits angiogenic sprouting; ectopic DLC1 expression rescues migration and sprouting in YAP-depleted cells.","method":"Chromatin immunoprecipitation (YAP/TAZ-TEAD binding to DLC1 promoter), siRNA depletion, rescue overexpression, live imaging of migration, sprouting assay, traction force microscopy","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — ChIP demonstrating direct transcriptional regulation, siRNA + rescue, multiple orthogonal functional assays","pmids":["31964713"],"is_preprint":false},{"year":2017,"finding":"DLC1 localizes asymmetrically to the cytoplasm at the cell front in avian trunk neural crest cells. This asymmetric localization depends on association of DLC1 with NEDD9. Asymmetric DLC1 creates differential RhoA activity (high at rear, fluctuating at front), determining polarized morphology and directional migration. SOX10 regulates DLC1 expression; SOX9 regulates NEDD9.","method":"RhoA FRET biosensor in vivo and in vitro, DLC1 overexpression/knockdown, NEDD9 co-immunoprecipitation and knockdown, in vivo neural crest imaging","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Moderate — in vivo FRET biosensor, co-IP, genetic knockdown in vivo, multiple orthogonal methods","pmids":["29084958"],"is_preprint":false},{"year":2022,"finding":"The co-crystal structure of the p120RasGAP SH3 domain bound directly to DLC1 RhoGAP shows the SH3 domain binds at a site partially overlapping the RhoA binding site and impinging on the catalytic arginine finger, directly inhibiting GAP activity. Mutations at this interface biochemically relieve SH3-mediated inhibition of DLC1 RhoGAP activity.","method":"X-ray co-crystallography, in vitro RhoGAP activity assay, mutagenesis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with mutagenesis validated by in vitro activity assay","pmids":["35970859"],"is_preprint":false},{"year":2005,"finding":"DLC1 knockout mice (DLC1-/-, Arhgap7) are embryonic lethal by day 10.5 post coitum, with defects in neural tube, brain, heart, and placenta. Cultured DLC1-deficient embryonic fibroblasts display alterations in actin filament organization and focal adhesions, demonstrating an essential developmental role in cytoskeletal regulation.","method":"Homologous recombination gene knockout, histological analysis, immunofluorescence (actin/focal adhesions) in primary fibroblasts","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean knockout mouse with defined phenotype, replicated in independent mouse model (PMID 20199662)","pmids":["15710412","20199662"],"is_preprint":false},{"year":2017,"finding":"Dlc1 is required for white and brown adipocyte differentiation. Dlc1 knockdown reduces lipid droplet formation and fat marker gene expression in white adipocytes, and reduces brown fat-specific gene expression and mitochondrial respiration in brown adipocytes. Dlc1-/- MEFs cannot differentiate into adipocytes, but this is rescued by ROCK and F-actin inhibitors, implicating the Rho pathway. PPARγ binds the Dlc1 promoter to regulate its expression during adipogenesis.","method":"siRNA knockdown, Dlc1-/- MEFs, ROCK/F-actin inhibitor rescue, ChIP (PPARγ binding), adipocyte differentiation assays, mitochondrial respiration measurement","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Moderate — KO cells with pathway rescue, ChIP, multiple orthogonal assays, single lab","pmids":["28358928"],"is_preprint":false},{"year":2020,"finding":"DLC1 negatively regulates TCTP (translationally controlled tumor protein) in a RhoGAP-independent manner. DLC1 and TCTP colocalize at focal adhesions and form a complex. Depletion of DLC1 increases TCTP expression; transfection with either WT or GAP-dead DLC1 (R718A) decreases TCTP levels. DLC1/TCTP interaction modulates Cdc42-JNK/NF-κB and N-WASP signaling to regulate cancer cell migration.","method":"Co-immunoprecipitation, colocalization imaging, GAP-dead mutant, siRNA knockdown, western blot, cell migration assay","journal":"Food & function","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, GAP-dead mutant establishes independence of GAP activity, multiple readouts, single lab","pmids":["33150340"],"is_preprint":false},{"year":2004,"finding":"p122 (rat DLC1 ortholog) localizes to caveolin-enriched membrane domains (caveolae) via its C-terminal GAP domain-containing region. Expression of p122/RhoGAP causes internalization of caveolin-1, and the GAP domain is responsible for its patchy plasma membrane distribution. This localization and function are cholesterol-dependent.","method":"EGFP tagging and live/fixed cell imaging, sucrose density gradient fractionation, immunostaining, cholesterol depletion","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — subcellular fractionation and imaging with functional consequence (caveolin internalization), single lab","pmids":["14723705"],"is_preprint":false},{"year":2008,"finding":"Focal adhesion localization of DLC1 (START-GAP1) is mediated by residues 265–459 ('FAT domain'). Expression of the FAT domain alone as a dominant-negative disrupts endogenous DLC1 localization, reduces cell migration, and impairs cell spreading. Focal adhesion localization is required for DLC1-mediated cell morphology changes.","method":"Immunofluorescence, deletion mutagenesis, dominant-negative expression, cell migration assay","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mutagenesis with dominant-negative approach and functional readouts, single lab","pmids":["19170769"],"is_preprint":false},{"year":2007,"finding":"Restoration of DLC1 expression in DLC1-null hepatocellular carcinoma cells causes inhibition of cell proliferation, disassembly of stress fibers, membrane protrusion, inhibition of cell migration, and dephosphorylation of focal adhesion proteins FAK, p130Cas, and paxillin.","method":"Adenoviral DLC1 transduction, cell proliferation assay, immunofluorescence, western blot of phospho-focal adhesion proteins, migration assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — adenoviral re-expression with multiple phenotypic readouts, single lab","pmids":["17292327"],"is_preprint":false},{"year":2010,"finding":"DLC1 silencing in non-malignant prostate epithelial cells does not promote more aggressive tumor phenotypes but promotes pro-angiogenic responses through VEGF upregulation. DLC1 loss leads to accumulation and nuclear localization of HIF-1α. VEGF modulation by DLC1 loss is dependent on EGFR-MEK-HIF-1α signaling, not RhoA.","method":"shRNA knockdown, VEGF ELISA, HIF-1α immunofluorescence/fractionation, EGFR/MEK inhibitors, RhoA inhibitor comparison","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA knockdown with pathway inhibitor dissection, multiple readouts, single lab","pmids":["20861185"],"is_preprint":false},{"year":2011,"finding":"Silencing of DLC1 in normal prostate epithelial cells reduces (not increases) cell migration due to upregulation of plasminogen activator inhibitor 1 (PAI-1). PAI-1 silencing rescues the migration defect. DLC1-K714E (GAP-inactive mutant) cannot decrease PAI-1 or rescue migration, indicating GAP-dependent PAI-1 regulation. DLC1-Y442F (tensin-binding defective) suppresses PAI-1 but does not restore migration, defining two independent DLC1 functions in normal cells.","method":"shRNA knockdown, DLC1 mutant re-expression, PAI-1 western blot, Transwell and wound-healing migration assays, PAI-1 siRNA rescue","journal":"Molecular cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA with domain-specific mutants, siRNA rescue, multiple assays, single lab","pmids":["22064653"],"is_preprint":false},{"year":2016,"finding":"Dlc1 interacts with non-muscle myosin heavy chain II-A (Myh9) in multiprotein complexes. Dlc1 overexpression increases phosphorylation of Myh9 and activates Rac1 GTPase, contributing to induced cell elongation morphology.","method":"Mass spectrometry interactome, co-immunoprecipitation validation, western blot for Myh9 phosphorylation, Rac1 activity pulldown","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP validation of MS interactome hit, functional readouts, single lab","pmids":["26977077"],"is_preprint":false},{"year":2007,"finding":"DLC1 protein undergoes nuclear translocation in a fraction of cells; this process requires the RhoGAP domain and a bipartite nuclear localization sequence. Nuclear DLC1 functions as an inducer of caspase-3-dependent apoptosis, while cytoplasmic DLC1 inhibits tumor cell proliferation and migration.","method":"DLC1 mutant analysis, immunofluorescence/subcellular fractionation, caspase-3 assay, cell migration/proliferation assays","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mutagenesis with functional consequences, subcellular localization with mechanistic link, single lab","pmids":["17888903"],"is_preprint":false},{"year":2008,"finding":"DLC1 tumor suppressor protein, when reintroduced into hepatoma cells with low DLC1 levels, reduces levels of GTP-bound RhoA; enforced expression of constitutively active RhoA mimics DLC1 loss in promoting hepatocellular carcinogenesis; down-regulation of RhoA selectively inhibits tumor growth of DLC1-disabled hepatoma cells, establishing DLC1 acts through the RhoA pathway in vivo.","method":"RNAi knockdown, constitutively active RhoA expression, shRNA RhoA knockdown, in vivo mouse hepatocellular carcinoma model","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with constitutively active and dominant-negative alleles in vivo and in vitro, multiple labs/groups converging","pmids":["18519636"],"is_preprint":false}],"current_model":"DLC1 is a multidomain focal adhesion protein (containing SAM, RhoGAP, and START domains) that functions as a RhoGAP with specificity for RhoA, RhoB, and RhoC (and limited Cdc42 activity); its RhoGAP activity is regulated by autoinhibition (SAM domain suppresses the RhoGAP domain), relieved by CDK5 phosphorylation, tensin3/PTEN binding, and PKA-induced dimerization, and inhibited by AKT phosphorylation, 14-3-3 protein binding, and p120RasGAP SH3 domain interaction; DLC1 localizes to focal adhesions via an LD-like motif that binds talin (in a force-sensitive manner dependent on talin R8 unfolding) and FAK, and via a FAT domain; its tumor suppressor activity requires both RhoGAP-dependent mechanisms (reducing RhoA-GTP, disrupting actin stress fibers and focal adhesions, suppressing Rho/ROCK/MLC2 signaling) and RhoGAP-independent mechanisms (START domain binding to caveolin-1, SAM domain binding to EF1A1, central domain binding to S100A10 to promote its degradation, α-catenin binding to stabilize adherens junctions); DLC1 is also a direct transcriptional target of YAP/TAZ-TEAD and is subject to post-translational regulation by CRL4A-FBXW5-mediated ubiquitin-proteasomal degradation; loss of DLC1 is embryonic lethal in mice due to defects in actin organization and development, and in cancer cells promotes RhoA-dependent metastasis including via TGF-β/SMAD3/PTHLH-driven bone metastasis."},"narrative":{"mechanistic_narrative":"DLC1 is a multidomain focal adhesion protein that functions as a tumor suppressor by acting as a GTPase-activating protein (GAP) for the Rho subfamily, with strong activity toward RhoA, RhoB, and RhoC and only limited activity toward Cdc42 [PMID:14633684, PMID:17932950]. Through this RhoGAP activity it lowers RhoA-GTP, disassembles actin stress fibers and focal adhesions, and suppresses downstream Rho/ROCK/MLC2 signaling, and these effects require its catalytic arginine finger [PMID:18648664, PMID:18519636]. DLC1 RhoGAP activity is held in a closed, autoinhibited conformation in which an N-terminal region (including the SAM domain) folds back onto the RhoGAP domain [PMID:18786931, PMID:25452387]; this autoinhibition is relieved by CDK5 phosphorylation, by binding of tensin family C2/actin-binding domains and PTEN, by PI(4,5)P2 engagement of a polybasic region, and by PKA-driven Ser549 phosphorylation that promotes activating dimerization, whereas AKT phosphorylation, 14-3-3 binding, and the p120RasGAP SH3 domain hold or convert DLC1 into the inactive state [PMID:19066281, PMID:19151751, PMID:19710422, PMID:22307599, PMID:23511482, PMID:25452387, PMID:29114068, PMID:31806702, PMID:35970859]. DLC1 is recruited to focal adhesions through an LD-like motif that binds talin and FAK and through a FAT domain, with talin R8 unfolding under mechanical force acting as a force-sensitive scaffold for downstream RhoA signaling [PMID:21969587, PMID:27265849, PMID:30028837, PMID:19170769]. Beyond catalysis, DLC1 also acts through RhoGAP-independent routes — START-domain binding to caveolin-1, SAM-domain binding to EF1A1, displacement of S100A10 to promote its degradation, and complex formation with α-catenin to stabilize adherens junctions [PMID:19158340, PMID:21372205, PMID:22473989, PMID:22693251]. Its expression is controlled by YAP/TAZ-TEAD transcription and its protein level by CRL4A-DDB1-FBXW5-mediated ubiquitin-proteasomal degradation [PMID:24082123, PMID:31964713]. DLC1 is essential for development, as its loss is embryonic lethal in mice with defective actin organization, and its loss in cancer cells drives RhoA-dependent metastasis, including TGF-β/SMAD3/PTHLH-driven osteolytic bone metastasis [PMID:24590291, PMID:15710412, PMID:20199662].","teleology":[{"year":2003,"claim":"Established the core molecular identity of DLC1 as a Rho-family GAP, defining the enzymatic activity around which all later mechanism was built.","evidence":"In vitro GAP activity assay showing specificity for RhoA and Cdc42","pmids":["14633684"],"confidence":"High","gaps":["Cellular substrate selectivity not yet resolved","No information on regulation of the activity"]},{"year":2008,"claim":"Resolved the substrate spectrum and revealed that flanking domains autoinhibit the catalytic domain, reframing DLC1 as a conformationally regulated enzyme.","evidence":"In vitro GAP assays comparing full-length versus isolated RhoGAP domain plus RhoA biosensor imaging; SAM-domain truncation/missense mutagenesis","pmids":["17932950","18786931"],"confidence":"High","gaps":["Trigger that relieves SAM autoinhibition not identified","Mechanism of focal adhesion recruitment independent of SAM/START not defined"]},{"year":2008,"claim":"Connected DLC1 catalytic activity to a defined cytoskeletal signaling output, showing it suppresses the Rho/ROCK/MLC2 axis in a GAP-dependent manner.","evidence":"Immunofluorescence, western blot, RhoGAP-dead (K714E) mutant, and dominant-active ROCK epistasis in hepatocellular carcinoma cells","pmids":["18648664"],"confidence":"High","gaps":["Did not address RhoGAP-independent contributions","In vivo relevance not tested in this study"]},{"year":2009,"claim":"Identified multiple inputs that tune DLC1 activity and localization — negative (14-3-3, p120RasGAP SH3) and positive (PI(4,5)P2, tensin binding) — establishing DLC1 as a signaling hub.","evidence":"Co-IP, in vitro GAP assays with purified proteins and PI(4,5)P2 liposomes, mutagenesis of binding motifs, RhoA pulldowns","pmids":["19066281","19151751","19710422","19826001","19440389"],"confidence":"High","gaps":["How these regulators are integrated in time and space unresolved","Some interactions characterized in single labs without reciprocal validation"]},{"year":2009,"claim":"Demonstrated that DLC1 suppresses migration through RhoGAP-independent protein interactions, beginning the dissection of catalytic versus scaffolding functions.","evidence":"NMR structure of the SAM domain, mass spectrometry identification of EF1A1, mutagenesis of a hydrophobic patch, and migration assays","pmids":["19158340"],"confidence":"High","gaps":["Quantitative contribution of EF1A1 axis versus RhoGAP activity unclear","Mechanism linking EF1A1 redistribution to migration not fully defined"]},{"year":2011,"claim":"Expanded the RhoGAP-independent tumor suppressor repertoire by showing DLC1 destabilizes S100A10 to limit plasminogen activation.","evidence":"Reciprocal co-IP, Annexin 2 competition assay, RhoGAP-independence controls, and invasion/migration assays","pmids":["21372205"],"confidence":"High","gaps":["Identity of the ubiquitin ligase for S100A10 not established","In vivo relevance of this axis not demonstrated"]},{"year":2011,"claim":"Defined the structural basis of DLC1 focal adhesion targeting through an LD-like motif binding talin and FAK, linking localization to tumor suppression.","evidence":"Co-IP, mutagenesis, focal adhesion imaging, and growth/colony-formation assays","pmids":["21969587"],"confidence":"High","gaps":["Functional separation of talin versus FAK binding not fully resolved","How localization potentiates GAP output mechanistically unclear"]},{"year":2012,"claim":"Established additional RhoGAP-independent (caveolin-1, α-catenin) and activating (tensin3) mechanisms, clarifying how distinct domains drive distinct anti-tumor outputs.","evidence":"Reciprocal co-IP with domain mapping, in vitro RhoGAP assays, FRAP, active Rho pulldown, and functional growth/junction assays","pmids":["22693251","22473989","22307599"],"confidence":"High","gaps":["Relative weighting of GAP-dependent and GAP-independent mechanisms in tumors unclear","Whether these complexes coexist on the same DLC1 molecule unknown"]},{"year":2013,"claim":"Defined opposing post-translational controls on DLC1 — activating PKA phosphorylation/dimerization and inactivating CRL4A-FBXW5 degradation — explaining DLC1 loss in cancer.","evidence":"Kinase assay, dimerization co-IP, ubiquitination assay, siRNA epistasis, RhoA pulldown, and tumor suppression assays","pmids":["23511482","24082123","23376848"],"confidence":"High","gaps":["Crosstalk between phospho-activation and ubiquitin-driven turnover not integrated","Upstream signals controlling FBXW5 recruitment unknown"]},{"year":2014,"claim":"Identified the kinase switches (CDK5 activating, AKT inhibiting) that toggle DLC1 between open dimeric active and closed monomeric inactive states, and linked DLC1 loss to bone metastasis.","evidence":"Kinase assays, autoinhibition co-IP, mutagenesis, focal adhesion imaging, and in vivo metastasis model with ROCK inhibition","pmids":["25452387","29114068","24590291"],"confidence":"High","gaps":["How CDK5 and AKT phosphorylation are coordinated in vivo unresolved","Therapeutic window of AKT inhibition in DLC1-positive tumors not defined here"]},{"year":2016,"claim":"Provided atomic-resolution definition of the talin R8–DLC1 LD interaction, anchoring the recruitment model structurally.","evidence":"X-ray crystallography of talin R8–DLC1 LD complex with interface mutagenesis validated in cells","pmids":["27265849"],"confidence":"High","gaps":["Competition with paxillin LD motifs for the same R8 surface in cells not quantified","Dynamics of binding under load not addressed by static structure"]},{"year":2018,"claim":"Established DLC1 as part of a mechanotransduction circuit in which force-induced talin R8 unfolding gates DLC1 signaling, and revealed a GAP-independent role in endothelial ICAM-1 adhesome assembly.","evidence":"Force-resistant talin mutant with AFM and traction force microscopy; ICAM-1 clustering, co-IP, and leukocyte adhesion assays","pmids":["30028837","30231995"],"confidence":"High","gaps":["How unfolding-dependent recruitment is converted to local RhoA inactivation mechanistically unclear","ICAM-1 adhesome role characterized in a single lab"]},{"year":2019,"claim":"Refined the activation mechanism by mapping SAM-domain engagement of tensin3 and PTEN C2 domains and demonstrating peptide-based reactivation of DLC1.","evidence":"Co-IP, in vitro RhoGAP assays, and cell-penetrating C2-derived peptide functional assays","pmids":["31806702"],"confidence":"High","gaps":["Structural basis of SAM–C2 binding not resolved","Specificity and off-target effects of the cyclic peptide not fully characterized"]},{"year":2020,"claim":"Placed DLC1 within mechanosensitive transcriptional control by identifying it as a direct YAP/TAZ-TEAD target governing endothelial migration and angiogenic sprouting.","evidence":"ChIP of YAP/TAZ-TEAD on the DLC1 promoter, siRNA depletion with rescue, traction force microscopy, and sprouting assays","pmids":["31964713"],"confidence":"High","gaps":["Feedback between DLC1-controlled mechanics and YAP/TAZ activity not resolved","Generalizability beyond endothelial context untested in this study"]},{"year":2022,"claim":"Provided the structural mechanism for p120RasGAP SH3 inhibition, showing it occludes the RhoA-binding site and catalytic arginine finger.","evidence":"Co-crystal structure of p120RasGAP SH3–DLC1 RhoGAP with interface mutagenesis and in vitro activity assays","pmids":["35970859"],"confidence":"High","gaps":["Cellular conditions selecting SH3-bound versus active state unknown","Interplay with other regulators at the same surface not defined"]},{"year":null,"claim":"How the many competing activating and inhibitory inputs are integrated spatiotemporally on a single DLC1 molecule to set local RhoA activity in a given cellular context remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No unified model reconciling kinase, lipid, scaffold, and ubiquitin inputs","Limited structural data on full-length autoinhibited DLC1","Relative in vivo contribution of GAP-dependent versus GAP-independent functions across tissues unclear"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,2,28]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,5,14,28]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,29,34]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[11,13,15]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[7]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[11,13,38]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[13,32]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4,38]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,18,39,22]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[16,20,39]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[29,27,30]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[26,30]}],"complexes":["focal adhesion","adherens junction (E-cadherin/β-catenin/α-catenin)","ICAM-1 adhesome"],"partners":["TLN1","PTK2","RASA1","TNS3","PTEN","CAV1","CTNNA1","S100A10"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q96QB1","full_name":"Rho GTPase-activating protein 7","aliases":["Deleted in liver cancer 1 protein","DLC-1","HP protein","Rho-type GTPase-activating protein 7","START domain-containing protein 12","StARD12","StAR-related lipid transfer protein 12"],"length_aa":1528,"mass_kda":170.6,"function":"Functions as a GTPase-activating protein for the small GTPases RHOA, RHOB, RHOC and CDC42, terminating their downstream signaling. This induces morphological changes and detachment through cytoskeletal reorganization, playing a critical role in biological processes such as cell migration and proliferation. Also functions in vivo as an activator of the phospholipase PLCD1. Active DLC1 increases cell migration velocity but reduces directionality. Required for growth factor-induced epithelial cell migration; in resting cells, interacts with TNS3 while PTEN interacts with the p85 regulatory subunit of the PI3K kinase complex but growth factor stimulation induces phosphorylation of TNS3 and PTEN, causing them to change their binding preference so that PTEN interacts with DLC1 and TNS3 interacts with p85 (PubMed:26166433). The PTEN-DLC1 complex translocates to the posterior of migrating cells to activate RHOA while the TNS3-p85 complex translocates to the leading edge of migrating cells to promote RAC1 activation (PubMed:26166433)","subcellular_location":"Cytoplasm; Cell junction, focal adhesion; Membrane","url":"https://www.uniprot.org/uniprotkb/Q96QB1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DLC1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DLC1","total_profiled":1310},"omim":[{"mim_id":"610533","title":"WW AND C2 DOMAINS-CONTAINING PROTEIN 1; WWC1","url":"https://www.omim.org/entry/610533"},{"mim_id":"609866","title":"START DOMAIN-CONTAINING PROTEIN 13; STARD13","url":"https://www.omim.org/entry/609866"},{"mim_id":"608942","title":"DYNEIN, LIGHT CHAIN, LC8 TYPE, 2; DYNLL2","url":"https://www.omim.org/entry/608942"},{"mim_id":"606266","title":"BCL2-MODIFYING FACTOR; BMF","url":"https://www.omim.org/entry/606266"},{"mim_id":"604258","title":"DLC1 RHO GTPase ACTIVATING PROTEIN; DLC1","url":"https://www.omim.org/entry/604258"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Endoplasmic reticulum","reliability":"Additional"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/DLC1"},"hgnc":{"alias_symbol":["HP","ARHGAP7","STARD12","DLC-1","p122-RhoGAP"],"prev_symbol":[]},"alphafold":{"accession":"Q96QB1","domains":[{"cath_id":"1.10.287.2070","chopping":"447-518","consensus_level":"medium","plddt":84.6142,"start":447,"end":518},{"cath_id":"1.10.555.10","chopping":"1021-1053_1074-1241_1262-1301","consensus_level":"medium","plddt":87.2215,"start":1021,"end":1301},{"cath_id":"3.30.530.20","chopping":"1308-1518","consensus_level":"high","plddt":87.8307,"start":1308,"end":1518}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96QB1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96QB1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96QB1-F1-predicted_aligned_error_v6.png","plddt_mean":55.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DLC1","jax_strain_url":"https://www.jax.org/strain/search?query=DLC1"},"sequence":{"accession":"Q96QB1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96QB1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96QB1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96QB1"}},"corpus_meta":[{"pmid":"14633684","id":"PMC_14633684","title":"Genetic 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assay, replicated across multiple labs in subsequent studies\",\n      \"pmids\": [\"14633684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Full-length DLC1 exhibits strong GAP activity for RhoA, RhoB, and RhoC but only very limited activity for Cdc42 in vitro; in contrast, the isolated RhoGAP domain showed 5- to 20-fold enhanced activity for RhoA, RhoB, RhoC, and Cdc42, suggesting the flanking domains impose autoinhibition. DLC1 reduces RhoA activity at the leading edge of cellular protrusions, and its anti-tumor activity involves both RhoGAP-dependent and RhoGAP-independent mechanisms. Unlike rat p122RhoGAP, human DLC1 was not capable of activating phospholipase C-delta1.\",\n      \"method\": \"In vitro GAP activity assay, RhoA biosensor live imaging, cell-based assays\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro biochemical assay plus live-cell biosensor imaging, multiple orthogonal methods in single study\",\n      \"pmids\": [\"17932950\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"DLC1 negatively regulates the Rho/ROCK/MLC2 pathway in hepatocellular carcinoma. Ectopic DLC1 expression abrogates Rho/ROCK-mediated cytoskeletal reorganization (stress fibers, focal adhesions) and downregulates cortical phosphorylation of myosin light chain 2 (MLC2) and MYPT1 phosphorylation at Thr853. A RhoGAP-deficient mutant (K714E) abolishes these inhibitory effects, confirming RhoGAP-dependency. Dominant-active ROCK rescues cells from DLC1-induced cytoskeletal collapse.\",\n      \"method\": \"Immunofluorescence, western blot, dominant-active ROCK epistasis, RhoGAP-dead mutant\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (immunofluorescence, western blot, mutant rescue, epistasis), single lab\",\n      \"pmids\": [\"18648664\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The SAM domain of DLC1 functions as an autoinhibitory domain of intrinsic RhoGAP activity. The SAM and START domains are dispensable for DLC1 association with focal adhesions. A dominant-negative N-terminal fragment displaces endogenous DLC1 from focal adhesions and profoundly inhibits cell migration.\",\n      \"method\": \"Structure-function analysis with truncation and missense mutants, cell morphology and migration assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic domain mutagenesis combined with functional assays, multiple orthogonal readouts\",\n      \"pmids\": [\"18786931\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"DLC1 interacts with 14-3-3 adaptor proteins via phosphoserine recognition motifs involving Ser327 and Ser431. Phorbol-ester-induced activation of PKC/PKD stimulates this association. Binding to 14-3-3 inhibits DLC1 GAP activity, facilitates RhoA signaling, and blocks DLC1 nucleocytoplasmic shuttling by masking a nuclear localization sequence.\",\n      \"method\": \"Co-immunoprecipitation, in vitro GAP assay, site-directed mutagenesis, subcellular fractionation/imaging\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — co-IP, in vitro GAP activity, mutagenesis of binding motifs, multiple orthogonal methods\",\n      \"pmids\": [\"19066281\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"p120Ras-GAP (RASA1) interacts with DLC1 and colocalizes in focal adhesions. The Ras-GAP SH3 domain binds the DLC1 RhoGAP domain and inhibits DLC1 RhoGAP activity in vitro. Overexpression of Ras-GAP impairs DLC1 growth-suppressing activity and increases RhoA activity in vivo.\",\n      \"method\": \"Co-immunoprecipitation, in vitro GAP assay with purified proteins, cell growth assay, active RhoA pulldown\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro reconstitution with purified proteins plus cellular functional assays, multiple orthogonal methods\",\n      \"pmids\": [\"19151751\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"The SAM domain of DLC1 binds eukaryotic elongation factor 1A1 (EF1A1) but not the SAM domain of DLC2. The solution structure of DLC1 SAM reveals a monomeric fold with four parallel helices. A hydrophobic patch (F38, L39, F40) is required for EF1A1 interaction. SAM-EF1A1 interaction facilitates EF1A1 distribution to membrane periphery upon growth factor stimulation and contributes to DLC1-mediated suppression of cell migration independent of the RhoGAP domain.\",\n      \"method\": \"Protein precipitation, mass spectrometry, NMR structure, mutagenesis, cell migration assay\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure plus mutagenesis plus functional assays, multiple orthogonal methods in single study\",\n      \"pmids\": [\"19158340\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"DLC1 activation requires binding to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] through a polybasic region (PBR) adjacent to the RhoGAP domain. PI(4,5)P2-containing membranes stimulate DLC1 GAP activity in vitro. A PBR-deficient DLC1 mutant is severely compromised in suppressing cell spreading, directed migration, and proliferation.\",\n      \"method\": \"Lipid-binding assay, in vitro GAP activity assay with PI(4,5)P2 liposomes, mutagenesis, cell-based assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with lipids, GAP activity assay, mutagenesis, multiple functional readouts\",\n      \"pmids\": [\"19710422\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Tensin1 requires its SH2 domain (R1488) to bind DLC1; tensin1 F302A mutation (abrogating PP1alpha binding) also reduces DLC1 association, linking PP1alpha and DLC1 to tensin1 signaling. DLC1 binding via tensin mediates Rho regulation, but PP1alpha has additional DLC1-independent effects on migration and invasion.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis, cell migration and invasion assays, MLC20 phosphorylation western blot\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with mutagenesis, multiple functional readouts, single lab\",\n      \"pmids\": [\"19826001\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Tensin2 binds DLC1 via its PTB domain at a novel binding site (residues 375–385 of DLC1). Deletion of this PTB-binding site partially reduces DLC1 RhoGAP activity and attenuates growth-suppressive activity without affecting focal adhesion localization or tensin1/cten interactions.\",\n      \"method\": \"Co-immunoprecipitation, deletion mutagenesis, RhoGAP activity assay, growth suppression assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with deletion mutants, RhoGAP activity assay, growth assay, single lab\",\n      \"pmids\": [\"19440389\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Tensin2 knockdown significantly reduces the ability of human foreskin fibroblasts to contract 3D collagen gels, associated with reduced Rho activity; this inhibition is reversed by depletion of DLC1, placing DLC1 downstream of tensin2 in regulating Rho-mediated actomyosin contraction and collagen remodeling.\",\n      \"method\": \"siRNA knockdown, 3D collagen gel contraction assay, RhoA activity pulldown, epistasis by double knockdown\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis by double knockdown with defined functional readout, single lab\",\n      \"pmids\": [\"20069572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DLC1 binds S100A10 (p11) in the cell cytoplasm; the interaction is mediated by central sequences in DLC1 and the C-terminus of S100A10. DLC1 competes with Annexin 2 for S100A10 binding, displacing S100A10 from Annexin 2 and making it accessible to ubiquitin-dependent degradation, thereby decreasing S100A10 steady-state levels and attenuating plasminogen activation. This mechanism inhibits cell migration, invasion, and anchorage-independent growth independently of DLC1 RhoGAP activity.\",\n      \"method\": \"Co-immunoprecipitation, colocalization imaging, competition assay, RhoGAP activity assay, cell invasion/migration assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, multiple orthogonal functional assays, competition biochemistry, single lab\",\n      \"pmids\": [\"21372205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DLC1 contains an 8-aa LD-like motif (residues 469LDDILYHV476) that is necessary for binding to talin and FAK. This motif is required for DLC1 localization to focal adhesions and for full tumor suppressor activity. FAK binding is independent of talin and vice versa. Mutants deficient in talin/FAK binding have impaired tumor suppressor activity despite retained ability to negatively regulate overall Rho-GTP.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis, focal adhesion localization imaging, bioassays (growth inhibition, colony formation)\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, mutagenesis, localization imaging, multiple functional assays, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"21969587\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"DLC1 forms a complex with α-catenin; binding is mediated by DLC1 N-terminal residues 340–435 and α-catenin residues 117–161. DLC1-α-catenin colocalize in cytosol and plasma membrane, where they associate with E-cadherin and β-catenin. DLC1-α-catenin complex reduces RhoA-GTP at the plasma membrane, increases E-cadherin mobility, and stabilizes adherens junctions. DLC1 GAP activity is required for α-catenin accumulation at the plasma membrane.\",\n      \"method\": \"Co-immunoprecipitation, mutagenesis, colocalization immunofluorescence, active Rho pulldown, E-cadherin FRAP, cell-based functional assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with domain mapping, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"22473989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Tensin3 activates DLC1 RhoGAP activity by binding DLC1 through its actin-binding domain, thereby releasing an autoinhibitory interaction between the SAM and RhoGAP domains of DLC1. Cten (C-terminal tensin-like protein), which lacks the actin-binding domain, does not activate DLC1. Tensin3 depletion augments actin stress fibers and focal adhesions and enhances cell motility via RhoA/ROCK signaling.\",\n      \"method\": \"Co-immunoprecipitation, in vitro RhoGAP activity assay, siRNA knockdown, actin/focal adhesion imaging, ROCK inhibitor rescue\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro RhoGAP activity assay plus cellular epistasis with multiple orthogonal methods\",\n      \"pmids\": [\"22307599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"DLC1 forms a complex with caveolin-1 (CAV-1); the interaction is mapped to the DLC1 START domain. Mutation of the START domain disrupts interaction and colocalization with CAV-1 and abolishes suppression of neoplastic growth, even though RhoA regulation is retained. This defines a RhoGAP-independent tumor suppressor mechanism mediated by the START domain-CAV-1 interaction.\",\n      \"method\": \"Co-immunoprecipitation, domain mutagenesis, colocalization imaging, anchorage-independent growth assay, active RhoA pulldown\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, domain mutagenesis, multiple functional assays, single lab\",\n      \"pmids\": [\"22693251\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DLC1 is ubiquitinated and degraded by the CRL4A (cullin 4A-RING ubiquitin ligase) complex via DDB1 and FBXW5 substrate receptor, providing a post-translational mechanism for DLC1 loss in NSCLC. siRNA suppression of cullin 4A, DDB1, or FBXW5 restores DLC1 protein, reduces RhoA-GTP, and causes DLC1-dependent decreases in NSCLC proliferation.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation (ubiquitin ligase complex), ubiquitination assay, active RhoA pulldown, cell proliferation assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP of ubiquitin ligase complex, siRNA epistasis, multiple functional readouts, single lab\",\n      \"pmids\": [\"24082123\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"PKA phosphorylates DLC1 at Ser549, enhancing its RhoGAP activity and promoting its tumor suppressor functions (suppression of hepatoma cell growth, motility, and metastasis). Ser549 phosphorylation induces dimerization of DLC1, and inducible dimerization alone can rescue tumor suppressive and RhoGAP activities of a Ser549-deletion DLC1 mutant.\",\n      \"method\": \"Site-directed mutagenesis, kinase assay, co-immunoprecipitation (dimerization), RhoGAP activity assay, in vitro and in vivo tumor suppression assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — kinase assay, mutagenesis, dimerization co-IP, in vitro RhoGAP activity, multiple functional assays\",\n      \"pmids\": [\"23511482\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"DLC1 expression in metastatic prostate carcinoma cells induces E-cadherin expression at the mRNA level through suppression of RhoA-GTP and RhoC-GTP via its RhoGAP function. Rho/ROCK inhibitors mimic this effect. Knockdown of RhoC (more than RhoA) increases E-cadherin. Constitutively active RhoA(V14) and RhoC(V14) could not be reversed by DLC1, confirming epistasis.\",\n      \"method\": \"Gene expression (RT-PCR/western blot), ROCK inhibitors, shRNA, constitutively active Rho mutants (epistasis), cell invasion assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with constitutively active mutants, RhoGAP-dependent mechanism validated, multiple orthogonal methods\",\n      \"pmids\": [\"23376848\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CDK5 is a major regulator of DLC1. CDK5 phosphorylates four serines in DLC1 N-terminal to the RhoGAP domain. When unphosphorylated, this N-terminal region acts as an autoinhibitory domain that binds the RhoGAP domain and places DLC1 in a closed, inactive conformation. CDK5 phosphorylation reduces this autoinhibitory binding and coordinately activates DLC1 localization to focal adhesions, RhoGAP activity, and ability to bind tensin and talin.\",\n      \"method\": \"Kinase assay (CDK5 phosphorylation), mutagenesis, co-immunoprecipitation (autoinhibitory domain interaction), focal adhesion localization imaging, RhoGAP activity assay, tensin/talin binding assay\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro kinase assay, co-IP autoinhibition, mutagenesis, multiple functional readouts, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"25452387\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Loss of DLC1 activates Rho-ROCK signaling, which mediates SMAD3 linker region phosphorylation and TGF-β-induced expression of parathyroid hormone-like hormone (PTHLH), leading to osteoclast maturation and osteolytic bone metastasis. Pharmacological inhibition of Rho-ROCK reduces PTHLH production and breast cancer bone metastasis in vitro and in vivo.\",\n      \"method\": \"siRNA knockdown, ROCK inhibitor, phosphorylation analysis, PTHLH expression assay, in vivo mouse metastasis model\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (knockdown, inhibitor, in vivo model), mechanistic pathway defined, single lab\",\n      \"pmids\": [\"24590291\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The crystal structure of the talin R8 domain bound to the DLC1 LD motif reveals that the DLC1 LD helix binds the four-helix bundle of talin R8 in a canonical triple-helix arrangement. The same R8 surface interacts with paxillin LD1 and LD2 motifs. Key charged residues stabilize R8-LD interactions, and mutations at this interface disrupt DLC1 binding and function in cells.\",\n      \"method\": \"X-ray crystallography, mutagenesis, in vitro binding assay, cell-based functional assays\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure plus mutagenesis validated in vitro and in cells\",\n      \"pmids\": [\"27265849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Multiple RTK ligands increase RhoA-GTP in cells via AKT activation. AKT phosphorylates three serines (S298, S329, S567) in DLC1 N-terminal to the RhoGAP domain, inducing strong binding of that N-terminal region to the RhoGAP domain and converting DLC1 from an open, active dimer to a closed, inactive monomer. This reduces RhoA-GTP hydrolysis, binding of other DLC1 ligands, colocalization with focal adhesions, and tumor suppressor activity. AKT inhibition has potent antitumor activity specifically in DLC1-positive cancers.\",\n      \"method\": \"Kinase assay, mutagenesis, co-immunoprecipitation, active RhoA pulldown, focal adhesion localization imaging, in vivo tumor model, isogenic DLC1+/- cell lines\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro kinase assay, mutagenesis, co-IP mechanism validation, multiple cell and in vivo models, multiple orthogonal methods\",\n      \"pmids\": [\"29114068\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Talin R8 domain unfolding by mechanical force is required for DLC1 downstream signaling. Using a talin mutant resistant to force-induced R8 unfolding, DLC1 signaling (RhoA regulation) and cell mechanics are dependent on talin unfolding status, defining a mechanotransduction mechanism where talin acts as a force-sensitive scaffold for DLC1.\",\n      \"method\": \"Protein engineering (force-resistant talin mutant), atomic force microscopy, traction force microscopy, biophysical assays\",\n      \"journal\": \"PLoS biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — protein engineering with biophysical validation (AFM), functional consequences measured, multiple orthogonal methods\",\n      \"pmids\": [\"30028837\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"DLC1 depletion in endothelial cells on stiff substrates reduces cell stiffness and impairs ICAM-1 adhesome formation by preventing recruitment of filamin B, α-actinin-4, and cortactin to clustered ICAM-1, thereby impairing leukocyte spreading. DLC1 overexpression rescues ICAM-1 adhesome stabilization. This function is independent of the DLC1 GAP domain.\",\n      \"method\": \"siRNA knockdown, DLC1 overexpression rescue, ICAM-1 clustering assay, co-immunoprecipitation, stiffness measurement (AFM), leukocyte adhesion assay\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA knockdown with rescue, functional readout, GAP-independence shown, single lab\",\n      \"pmids\": [\"30231995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The DLC1 SAM domain binds to specific peptide motifs within the C2 domains of tensin3 and PTEN. This SAM-C2 interaction mediates DLC1 activation. Peptides containing the C2 binding motifs block the C2-SAM interaction, promote DLC1 RhoGAP activity, decrease RhoA activation, and reduce tumor cell growth and migration. A cyclic version of the TNS3 C2-derived peptide enters cancer cells and effectively inhibits migration.\",\n      \"method\": \"Co-immunoprecipitation, in vitro RhoGAP activity assay, cell-penetrating peptide functional assays, soft agar growth, migration assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro biochemical reconstitution, RhoGAP activity assay, multiple functional cellular assays, single lab\",\n      \"pmids\": [\"31806702\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DLC1 is a direct transcriptional target of the activated YAP/TAZ-TEAD complex. Substrate stiffening and VEGF stimulate DLC1 expression in endothelial cells in a YAP/TAZ-dependent manner. DLC1 limits F-actin fiber formation, integrin-based focal adhesion lifetime, and traction forces. DLC1 depletion impairs endothelial cell polarization in directed collective migration and inhibits angiogenic sprouting; ectopic DLC1 expression rescues migration and sprouting in YAP-depleted cells.\",\n      \"method\": \"Chromatin immunoprecipitation (YAP/TAZ-TEAD binding to DLC1 promoter), siRNA depletion, rescue overexpression, live imaging of migration, sprouting assay, traction force microscopy\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP demonstrating direct transcriptional regulation, siRNA + rescue, multiple orthogonal functional assays\",\n      \"pmids\": [\"31964713\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"DLC1 localizes asymmetrically to the cytoplasm at the cell front in avian trunk neural crest cells. This asymmetric localization depends on association of DLC1 with NEDD9. Asymmetric DLC1 creates differential RhoA activity (high at rear, fluctuating at front), determining polarized morphology and directional migration. SOX10 regulates DLC1 expression; SOX9 regulates NEDD9.\",\n      \"method\": \"RhoA FRET biosensor in vivo and in vitro, DLC1 overexpression/knockdown, NEDD9 co-immunoprecipitation and knockdown, in vivo neural crest imaging\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo FRET biosensor, co-IP, genetic knockdown in vivo, multiple orthogonal methods\",\n      \"pmids\": [\"29084958\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The co-crystal structure of the p120RasGAP SH3 domain bound directly to DLC1 RhoGAP shows the SH3 domain binds at a site partially overlapping the RhoA binding site and impinging on the catalytic arginine finger, directly inhibiting GAP activity. Mutations at this interface biochemically relieve SH3-mediated inhibition of DLC1 RhoGAP activity.\",\n      \"method\": \"X-ray co-crystallography, in vitro RhoGAP activity assay, mutagenesis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with mutagenesis validated by in vitro activity assay\",\n      \"pmids\": [\"35970859\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"DLC1 knockout mice (DLC1-/-, Arhgap7) are embryonic lethal by day 10.5 post coitum, with defects in neural tube, brain, heart, and placenta. Cultured DLC1-deficient embryonic fibroblasts display alterations in actin filament organization and focal adhesions, demonstrating an essential developmental role in cytoskeletal regulation.\",\n      \"method\": \"Homologous recombination gene knockout, histological analysis, immunofluorescence (actin/focal adhesions) in primary fibroblasts\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean knockout mouse with defined phenotype, replicated in independent mouse model (PMID 20199662)\",\n      \"pmids\": [\"15710412\", \"20199662\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Dlc1 is required for white and brown adipocyte differentiation. Dlc1 knockdown reduces lipid droplet formation and fat marker gene expression in white adipocytes, and reduces brown fat-specific gene expression and mitochondrial respiration in brown adipocytes. Dlc1-/- MEFs cannot differentiate into adipocytes, but this is rescued by ROCK and F-actin inhibitors, implicating the Rho pathway. PPARγ binds the Dlc1 promoter to regulate its expression during adipogenesis.\",\n      \"method\": \"siRNA knockdown, Dlc1-/- MEFs, ROCK/F-actin inhibitor rescue, ChIP (PPARγ binding), adipocyte differentiation assays, mitochondrial respiration measurement\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO cells with pathway rescue, ChIP, multiple orthogonal assays, single lab\",\n      \"pmids\": [\"28358928\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"DLC1 negatively regulates TCTP (translationally controlled tumor protein) in a RhoGAP-independent manner. DLC1 and TCTP colocalize at focal adhesions and form a complex. Depletion of DLC1 increases TCTP expression; transfection with either WT or GAP-dead DLC1 (R718A) decreases TCTP levels. DLC1/TCTP interaction modulates Cdc42-JNK/NF-κB and N-WASP signaling to regulate cancer cell migration.\",\n      \"method\": \"Co-immunoprecipitation, colocalization imaging, GAP-dead mutant, siRNA knockdown, western blot, cell migration assay\",\n      \"journal\": \"Food & function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, GAP-dead mutant establishes independence of GAP activity, multiple readouts, single lab\",\n      \"pmids\": [\"33150340\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"p122 (rat DLC1 ortholog) localizes to caveolin-enriched membrane domains (caveolae) via its C-terminal GAP domain-containing region. Expression of p122/RhoGAP causes internalization of caveolin-1, and the GAP domain is responsible for its patchy plasma membrane distribution. This localization and function are cholesterol-dependent.\",\n      \"method\": \"EGFP tagging and live/fixed cell imaging, sucrose density gradient fractionation, immunostaining, cholesterol depletion\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — subcellular fractionation and imaging with functional consequence (caveolin internalization), single lab\",\n      \"pmids\": [\"14723705\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Focal adhesion localization of DLC1 (START-GAP1) is mediated by residues 265–459 ('FAT domain'). Expression of the FAT domain alone as a dominant-negative disrupts endogenous DLC1 localization, reduces cell migration, and impairs cell spreading. Focal adhesion localization is required for DLC1-mediated cell morphology changes.\",\n      \"method\": \"Immunofluorescence, deletion mutagenesis, dominant-negative expression, cell migration assay\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mutagenesis with dominant-negative approach and functional readouts, single lab\",\n      \"pmids\": [\"19170769\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Restoration of DLC1 expression in DLC1-null hepatocellular carcinoma cells causes inhibition of cell proliferation, disassembly of stress fibers, membrane protrusion, inhibition of cell migration, and dephosphorylation of focal adhesion proteins FAK, p130Cas, and paxillin.\",\n      \"method\": \"Adenoviral DLC1 transduction, cell proliferation assay, immunofluorescence, western blot of phospho-focal adhesion proteins, migration assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — adenoviral re-expression with multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"17292327\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"DLC1 silencing in non-malignant prostate epithelial cells does not promote more aggressive tumor phenotypes but promotes pro-angiogenic responses through VEGF upregulation. DLC1 loss leads to accumulation and nuclear localization of HIF-1α. VEGF modulation by DLC1 loss is dependent on EGFR-MEK-HIF-1α signaling, not RhoA.\",\n      \"method\": \"shRNA knockdown, VEGF ELISA, HIF-1α immunofluorescence/fractionation, EGFR/MEK inhibitors, RhoA inhibitor comparison\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA knockdown with pathway inhibitor dissection, multiple readouts, single lab\",\n      \"pmids\": [\"20861185\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Silencing of DLC1 in normal prostate epithelial cells reduces (not increases) cell migration due to upregulation of plasminogen activator inhibitor 1 (PAI-1). PAI-1 silencing rescues the migration defect. DLC1-K714E (GAP-inactive mutant) cannot decrease PAI-1 or rescue migration, indicating GAP-dependent PAI-1 regulation. DLC1-Y442F (tensin-binding defective) suppresses PAI-1 but does not restore migration, defining two independent DLC1 functions in normal cells.\",\n      \"method\": \"shRNA knockdown, DLC1 mutant re-expression, PAI-1 western blot, Transwell and wound-healing migration assays, PAI-1 siRNA rescue\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA with domain-specific mutants, siRNA rescue, multiple assays, single lab\",\n      \"pmids\": [\"22064653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Dlc1 interacts with non-muscle myosin heavy chain II-A (Myh9) in multiprotein complexes. Dlc1 overexpression increases phosphorylation of Myh9 and activates Rac1 GTPase, contributing to induced cell elongation morphology.\",\n      \"method\": \"Mass spectrometry interactome, co-immunoprecipitation validation, western blot for Myh9 phosphorylation, Rac1 activity pulldown\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP validation of MS interactome hit, functional readouts, single lab\",\n      \"pmids\": [\"26977077\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"DLC1 protein undergoes nuclear translocation in a fraction of cells; this process requires the RhoGAP domain and a bipartite nuclear localization sequence. Nuclear DLC1 functions as an inducer of caspase-3-dependent apoptosis, while cytoplasmic DLC1 inhibits tumor cell proliferation and migration.\",\n      \"method\": \"DLC1 mutant analysis, immunofluorescence/subcellular fractionation, caspase-3 assay, cell migration/proliferation assays\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mutagenesis with functional consequences, subcellular localization with mechanistic link, single lab\",\n      \"pmids\": [\"17888903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"DLC1 tumor suppressor protein, when reintroduced into hepatoma cells with low DLC1 levels, reduces levels of GTP-bound RhoA; enforced expression of constitutively active RhoA mimics DLC1 loss in promoting hepatocellular carcinogenesis; down-regulation of RhoA selectively inhibits tumor growth of DLC1-disabled hepatoma cells, establishing DLC1 acts through the RhoA pathway in vivo.\",\n      \"method\": \"RNAi knockdown, constitutively active RhoA expression, shRNA RhoA knockdown, in vivo mouse hepatocellular carcinoma model\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with constitutively active and dominant-negative alleles in vivo and in vitro, multiple labs/groups converging\",\n      \"pmids\": [\"18519636\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DLC1 is a multidomain focal adhesion protein (containing SAM, RhoGAP, and START domains) that functions as a RhoGAP with specificity for RhoA, RhoB, and RhoC (and limited Cdc42 activity); its RhoGAP activity is regulated by autoinhibition (SAM domain suppresses the RhoGAP domain), relieved by CDK5 phosphorylation, tensin3/PTEN binding, and PKA-induced dimerization, and inhibited by AKT phosphorylation, 14-3-3 protein binding, and p120RasGAP SH3 domain interaction; DLC1 localizes to focal adhesions via an LD-like motif that binds talin (in a force-sensitive manner dependent on talin R8 unfolding) and FAK, and via a FAT domain; its tumor suppressor activity requires both RhoGAP-dependent mechanisms (reducing RhoA-GTP, disrupting actin stress fibers and focal adhesions, suppressing Rho/ROCK/MLC2 signaling) and RhoGAP-independent mechanisms (START domain binding to caveolin-1, SAM domain binding to EF1A1, central domain binding to S100A10 to promote its degradation, α-catenin binding to stabilize adherens junctions); DLC1 is also a direct transcriptional target of YAP/TAZ-TEAD and is subject to post-translational regulation by CRL4A-FBXW5-mediated ubiquitin-proteasomal degradation; loss of DLC1 is embryonic lethal in mice due to defects in actin organization and development, and in cancer cells promotes RhoA-dependent metastasis including via TGF-β/SMAD3/PTHLH-driven bone metastasis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DLC1 is a multidomain focal adhesion protein that functions as a tumor suppressor by acting as a GTPase-activating protein (GAP) for the Rho subfamily, with strong activity toward RhoA, RhoB, and RhoC and only limited activity toward Cdc42 [#0, #1]. Through this RhoGAP activity it lowers RhoA-GTP, disassembles actin stress fibers and focal adhesions, and suppresses downstream Rho/ROCK/MLC2 signaling, and these effects require its catalytic arginine finger [#2, #39]. DLC1 RhoGAP activity is held in a closed, autoinhibited conformation in which an N-terminal region (including the SAM domain) folds back onto the RhoGAP domain [#3, #19]; this autoinhibition is relieved by CDK5 phosphorylation, by binding of tensin family C2/actin-binding domains and PTEN, by PI(4,5)P2 engagement of a polybasic region, and by PKA-driven Ser549 phosphorylation that promotes activating dimerization, whereas AKT phosphorylation, 14-3-3 binding, and the p120RasGAP SH3 domain hold or convert DLC1 into the inactive state [#4, #5, #7, #14, #17, #19, #22, #25, #28]. DLC1 is recruited to focal adhesions through an LD-like motif that binds talin and FAK and through a FAT domain, with talin R8 unfolding under mechanical force acting as a force-sensitive scaffold for downstream RhoA signaling [#12, #21, #23, #33]. Beyond catalysis, DLC1 also acts through RhoGAP-independent routes — START-domain binding to caveolin-1, SAM-domain binding to EF1A1, displacement of S100A10 to promote its degradation, and complex formation with \\u03b1-catenin to stabilize adherens junctions [#6, #11, #13, #15]. Its expression is controlled by YAP/TAZ-TEAD transcription and its protein level by CRL4A-DDB1-FBXW5-mediated ubiquitin-proteasomal degradation [#16, #26]. DLC1 is essential for development, as its loss is embryonic lethal in mice with defective actin organization, and its loss in cancer cells drives RhoA-dependent metastasis, including TGF-\\u03b2/SMAD3/PTHLH-driven osteolytic bone metastasis [#20, #29].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established the core molecular identity of DLC1 as a Rho-family GAP, defining the enzymatic activity around which all later mechanism was built.\",\n      \"evidence\": \"In vitro GAP activity assay showing specificity for RhoA and Cdc42\",\n      \"pmids\": [\"14633684\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular substrate selectivity not yet resolved\", \"No information on regulation of the activity\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Resolved the substrate spectrum and revealed that flanking domains autoinhibit the catalytic domain, reframing DLC1 as a conformationally regulated enzyme.\",\n      \"evidence\": \"In vitro GAP assays comparing full-length versus isolated RhoGAP domain plus RhoA biosensor imaging; SAM-domain truncation/missense mutagenesis\",\n      \"pmids\": [\"17932950\", \"18786931\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Trigger that relieves SAM autoinhibition not identified\", \"Mechanism of focal adhesion recruitment independent of SAM/START not defined\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Connected DLC1 catalytic activity to a defined cytoskeletal signaling output, showing it suppresses the Rho/ROCK/MLC2 axis in a GAP-dependent manner.\",\n      \"evidence\": \"Immunofluorescence, western blot, RhoGAP-dead (K714E) mutant, and dominant-active ROCK epistasis in hepatocellular carcinoma cells\",\n      \"pmids\": [\"18648664\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address RhoGAP-independent contributions\", \"In vivo relevance not tested in this study\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified multiple inputs that tune DLC1 activity and localization — negative (14-3-3, p120RasGAP SH3) and positive (PI(4,5)P2, tensin binding) — establishing DLC1 as a signaling hub.\",\n      \"evidence\": \"Co-IP, in vitro GAP assays with purified proteins and PI(4,5)P2 liposomes, mutagenesis of binding motifs, RhoA pulldowns\",\n      \"pmids\": [\"19066281\", \"19151751\", \"19710422\", \"19826001\", \"19440389\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How these regulators are integrated in time and space unresolved\", \"Some interactions characterized in single labs without reciprocal validation\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Demonstrated that DLC1 suppresses migration through RhoGAP-independent protein interactions, beginning the dissection of catalytic versus scaffolding functions.\",\n      \"evidence\": \"NMR structure of the SAM domain, mass spectrometry identification of EF1A1, mutagenesis of a hydrophobic patch, and migration assays\",\n      \"pmids\": [\"19158340\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative contribution of EF1A1 axis versus RhoGAP activity unclear\", \"Mechanism linking EF1A1 redistribution to migration not fully defined\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Expanded the RhoGAP-independent tumor suppressor repertoire by showing DLC1 destabilizes S100A10 to limit plasminogen activation.\",\n      \"evidence\": \"Reciprocal co-IP, Annexin 2 competition assay, RhoGAP-independence controls, and invasion/migration assays\",\n      \"pmids\": [\"21372205\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the ubiquitin ligase for S100A10 not established\", \"In vivo relevance of this axis not demonstrated\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the structural basis of DLC1 focal adhesion targeting through an LD-like motif binding talin and FAK, linking localization to tumor suppression.\",\n      \"evidence\": \"Co-IP, mutagenesis, focal adhesion imaging, and growth/colony-formation assays\",\n      \"pmids\": [\"21969587\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional separation of talin versus FAK binding not fully resolved\", \"How localization potentiates GAP output mechanistically unclear\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Established additional RhoGAP-independent (caveolin-1, \\u03b1-catenin) and activating (tensin3) mechanisms, clarifying how distinct domains drive distinct anti-tumor outputs.\",\n      \"evidence\": \"Reciprocal co-IP with domain mapping, in vitro RhoGAP assays, FRAP, active Rho pulldown, and functional growth/junction assays\",\n      \"pmids\": [\"22693251\", \"22473989\", \"22307599\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Relative weighting of GAP-dependent and GAP-independent mechanisms in tumors unclear\", \"Whether these complexes coexist on the same DLC1 molecule unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined opposing post-translational controls on DLC1 — activating PKA phosphorylation/dimerization and inactivating CRL4A-FBXW5 degradation — explaining DLC1 loss in cancer.\",\n      \"evidence\": \"Kinase assay, dimerization co-IP, ubiquitination assay, siRNA epistasis, RhoA pulldown, and tumor suppression assays\",\n      \"pmids\": [\"23511482\", \"24082123\", \"23376848\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Crosstalk between phospho-activation and ubiquitin-driven turnover not integrated\", \"Upstream signals controlling FBXW5 recruitment unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified the kinase switches (CDK5 activating, AKT inhibiting) that toggle DLC1 between open dimeric active and closed monomeric inactive states, and linked DLC1 loss to bone metastasis.\",\n      \"evidence\": \"Kinase assays, autoinhibition co-IP, mutagenesis, focal adhesion imaging, and in vivo metastasis model with ROCK inhibition\",\n      \"pmids\": [\"25452387\", \"29114068\", \"24590291\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CDK5 and AKT phosphorylation are coordinated in vivo unresolved\", \"Therapeutic window of AKT inhibition in DLC1-positive tumors not defined here\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Provided atomic-resolution definition of the talin R8–DLC1 LD interaction, anchoring the recruitment model structurally.\",\n      \"evidence\": \"X-ray crystallography of talin R8–DLC1 LD complex with interface mutagenesis validated in cells\",\n      \"pmids\": [\"27265849\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Competition with paxillin LD motifs for the same R8 surface in cells not quantified\", \"Dynamics of binding under load not addressed by static structure\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Established DLC1 as part of a mechanotransduction circuit in which force-induced talin R8 unfolding gates DLC1 signaling, and revealed a GAP-independent role in endothelial ICAM-1 adhesome assembly.\",\n      \"evidence\": \"Force-resistant talin mutant with AFM and traction force microscopy; ICAM-1 clustering, co-IP, and leukocyte adhesion assays\",\n      \"pmids\": [\"30028837\", \"30231995\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How unfolding-dependent recruitment is converted to local RhoA inactivation mechanistically unclear\", \"ICAM-1 adhesome role characterized in a single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Refined the activation mechanism by mapping SAM-domain engagement of tensin3 and PTEN C2 domains and demonstrating peptide-based reactivation of DLC1.\",\n      \"evidence\": \"Co-IP, in vitro RhoGAP assays, and cell-penetrating C2-derived peptide functional assays\",\n      \"pmids\": [\"31806702\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of SAM–C2 binding not resolved\", \"Specificity and off-target effects of the cyclic peptide not fully characterized\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Placed DLC1 within mechanosensitive transcriptional control by identifying it as a direct YAP/TAZ-TEAD target governing endothelial migration and angiogenic sprouting.\",\n      \"evidence\": \"ChIP of YAP/TAZ-TEAD on the DLC1 promoter, siRNA depletion with rescue, traction force microscopy, and sprouting assays\",\n      \"pmids\": [\"31964713\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Feedback between DLC1-controlled mechanics and YAP/TAZ activity not resolved\", \"Generalizability beyond endothelial context untested in this study\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Provided the structural mechanism for p120RasGAP SH3 inhibition, showing it occludes the RhoA-binding site and catalytic arginine finger.\",\n      \"evidence\": \"Co-crystal structure of p120RasGAP SH3–DLC1 RhoGAP with interface mutagenesis and in vitro activity assays\",\n      \"pmids\": [\"35970859\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cellular conditions selecting SH3-bound versus active state unknown\", \"Interplay with other regulators at the same surface not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the many competing activating and inhibitory inputs are integrated spatiotemporally on a single DLC1 molecule to set local RhoA activity in a given cellular context remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No unified model reconciling kinase, lipid, scaffold, and ubiquitin inputs\", \"Limited structural data on full-length autoinhibited DLC1\", \"Relative in vivo contribution of GAP-dependent versus GAP-independent functions across tissues unclear\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 28]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 5, 14, 28]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 29, 34]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [11, 13, 15]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005925\", \"supporting_discovery_ids\": [3, 12, 33, 21]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [11, 13, 38]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [13, 32]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4, 38]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 18, 39, 22]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [16, 20, 39]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [29, 27, 30]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [26, 30]}\n    ],\n    \"complexes\": [\"focal adhesion\", \"adherens junction (E-cadherin/\\u03b2-catenin/\\u03b1-catenin)\", \"ICAM-1 adhesome\"],\n    \"partners\": [\"TLN1\", \"PTK2\", \"RASA1\", \"TNS3\", \"PTEN\", \"CAV1\", \"CTNNA1\", \"S100A10\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}