{"gene":"WIPF1","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":1997,"finding":"WIP (WIPF1) was identified as a WASP-interacting protein via yeast two-hybrid; it coimmunoprecipitates with WASP from lymphocytes, binds WASP at a site distinct from the Cdc42 binding site, and has actin and profilin binding motifs. Expression of WIP in B cells induced actin polymerization and cerebriform projections; a WIP truncation mutant lacking the actin-binding motif failed to do so, establishing the actin-binding domain as required for this function.","method":"Yeast two-hybrid, co-immunoprecipitation, expression in human B cells with truncation mutant","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, domain mapping with truncation mutant, functional readout; foundational paper replicated across many subsequent studies","pmids":["9405671"],"is_preprint":false},{"year":1998,"finding":"WIP binds to the adaptor protein Nck via the second SH3 domain of Nck; the Nck-binding site on WIP (amino acids 321–415) is distinct from the WASP-binding site (amino acids 416–488). Profilin is found in Nck precipitates, suggesting Nck links to the cytoskeleton via WIP and profilin.","method":"GST pulldown with recombinant Nck from BJAB cell lysates, domain mapping with WIP fragments","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct pulldown with domain mapping, single lab, two orthogonal methods (precipitation + co-IP of profilin)","pmids":["9694849"],"is_preprint":false},{"year":1998,"finding":"The yeast WIP homologue End5p/verprolin interacts with the WASp homologue Las17p (yeast ortholog of WASP); high-copy LAS17 partially suppresses end5-1 growth and endocytosis defects, establishing that the WIP–WASP functional partnership is conserved in yeast and essential for endocytosis.","method":"Yeast two-hybrid, high-copy suppressor screen, endocytosis assay in yeast","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis (suppressor screen) plus two-hybrid binding, replicated concept in multiple organisms","pmids":["9742397"],"is_preprint":false},{"year":1999,"finding":"Human WIP functionally complements yeast vrp1 (verprolin) mutations, restoring cytoskeletal organization and endocytosis; this complementation requires the WH2 actin-binding domain and the profilin-binding domain of WIP, establishing these as the functional core of WIP's role in cell polarity and actin organization.","method":"Yeast complementation assay, WIP domain mutants, immunofluorescence localization in yeast","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — reconstituted function in yeast with domain mapping, direct localization, single lab","pmids":["10358064"],"is_preprint":false},{"year":2000,"finding":"WIP mediates recruitment of N-WASP to vaccinia virus actin tails via the WH1 domain of N-WASP (not the polyproline region). For Shigella, N-WASP recruits WIP. The N-WASP–WIP complex integrates signaling cascades (SH2/SH3-adaptor pathway and Cdc42 pathway) leading to Arp2/3-dependent actin polymerization and pathogen actin-based motility.","method":"Cell-based actin tail assays with domain mutants, co-localization, dominant-negative experiments","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple genetic/biochemical approaches, two distinct pathogen models, independently replicated concept","pmids":["10878810"],"is_preprint":false},{"year":2001,"finding":"WIP directly interacts with N-WASP and with actin; WIP retards N-WASP/Cdc42-activated actin polymerization by the Arp2/3 complex and stabilizes actin filaments. Microinjection of WIP induces filopodia in an N-WASP-dependent manner; anti-WIP antibody blocks filopodium induction by bradykinin, Cdc42(V12), and N-WASP, establishing WIP and N-WASP as a functional unit in filopodium formation.","method":"In vitro actin polymerization assay (pyrene-actin), microinjection with function-blocking antibodies, immunofluorescence","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro reconstitution assay plus microinjection functional experiments, multiple orthogonal methods","pmids":["11331876"],"is_preprint":false},{"year":2001,"finding":"Rat WIP co-immunoprecipitates with N-WASP in vivo and co-localizes with actin stress fibers. Co-expression of WIP and N-WASP redistributes N-WASP from the nucleus to perinuclear/actin-associated locations and dissolves stress fibers while promoting filopodia formation, indicating WIP controls N-WASP subcellular localization.","method":"Co-immunoprecipitation, immunofluorescence, co-expression in fibroblasts and tumor cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP plus localization imaging, single lab, two orthogonal methods","pmids":["11687573"],"is_preprint":false},{"year":2002,"finding":"NMR structure of the N-WASP EVH1 (WH1) domain in complex with a 25-residue WIP motif revealed a novel recognition mechanism: the WIP peptide wraps around the EVH1 domain contacting an extended surface, mechanistically explaining how WAS missense mutations in this domain disrupt WIP binding.","method":"NMR structure determination with WIP peptide","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — high-resolution NMR structure with direct functional correlation to disease mutations","pmids":["12437929"],"is_preprint":false},{"year":2002,"finding":"WIP-deficient mice have T cells that fail to proliferate, secrete IL-2, increase F-actin content, polarize, or form immune synapses after TCR ligation, establishing WIP as essential for T cell activation and immunological synapse formation. WIP-deficient B cells show enhanced proliferation, indicating differential roles in T vs B cells.","method":"WIP knockout mouse model, T cell proliferation/activation assays, F-actin staining, conjugate formation assay","journal":"Immunity","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic KO with multiple defined cellular phenotype readouts, replicated across several studies","pmids":["11869681"],"is_preprint":false},{"year":2002,"finding":"Phosphatidylinositol 4,5-bisphosphate (PIP2)-induced vesicle motility requires N-WASP and involves WIP recruitment alongside Nck and Grb2; reconstitution in N-WASP-defective cells with mutants showed that both the WH1 domain (which recruits WIP) and the polyproline domain contribute significantly to N-WASP recruitment/activation at vesicle surfaces.","method":"Reconstitution of vesicle motility in N-WASP-deficient cells with domain mutants, fluorescence microscopy","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — reconstitution assay with multiple domain mutants, single lab","pmids":["12147689"],"is_preprint":false},{"year":2003,"finding":"Cortactin SH3 domain interacts with WIP in an SH3-dependent manner (GST pulldown). WIP increases the efficiency of cortactin-mediated Arp2/3 complex activation in a concentration-dependent manner, and co-expression of cortactin and WIP stimulates membrane protrusions.","method":"Yeast two-hybrid, GST-cortactin pulldown, in vitro Arp2/3 actin polymerization assay, overexpression/localization","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro Arp2/3 assay plus binding studies and cellular readout, single lab multiple orthogonal methods","pmids":["12620186"],"is_preprint":false},{"year":2003,"finding":"WIP participates in PDGF-induced ruffle formation: overexpression enhances ruffling, microinjection of anti-WIP antibody or WIP deficiency decreases ruffling, and a WIP mutant lacking the actin-binding site blocks PDGF-induced membrane ruffling in murine fibroblasts, establishing WIP's actin-binding domain as required for ruffle formation downstream of PDGF/Rac1.","method":"Microinjection of anti-WIP antibody, WIP overexpression, domain mutant (lacking actin-binding site), immunofluorescence, video microscopy in 3T3 and WIP-/- fibroblasts","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — antibody microinjection, KO cells, and domain mutant with functional readout, multiple orthogonal approaches","pmids":["12724353"],"is_preprint":false},{"year":2004,"finding":"Toca-1 (a PCH/F-BAR protein) binds both N-WASP and Cdc42 and activates the N-WASP–WIP complex (the predominant form of N-WASP in cells) to promote actin nucleation; two distinct Cdc42 effectors (N-WASP–WIP and Toca-1) cooperate and are both required for Cdc42-induced actin assembly.","method":"Biochemical purification of Toca-1, in vitro actin assembly assays, co-IP, depletion experiments","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — biochemical purification plus in vitro reconstitution, multiple orthogonal methods, widely replicated","pmids":["15260990"],"is_preprint":false},{"year":2004,"finding":"WIP-deficient mast cells show impaired degranulation, IL-6 secretion, calcium mobilization, and reduced phosphorylation of Syk, PLCγ2, and JNK after FcεRI ligation. WIP co-immunoprecipitates with Syk after FcεRI ligation and inhibits Syk degradation, establishing WIP as a regulator of FcεRI signaling via maintenance of Syk levels.","method":"WIP-/- bone marrow-derived mast cells, degranulation assay, co-immunoprecipitation with Syk, immunoblotting for Syk levels","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO model with specific molecular mechanism (Syk association and stability), co-IP validation","pmids":["14757742"],"is_preprint":false},{"year":2006,"finding":"WIP is essential for podosome formation in dendritic cells (DCs): WIP-/- DCs cannot form actin cores containing WASP and cortactin. WIP regulates podosome structure by controlling calpain-mediated cleavage of WASP and by facilitating WASP localization to actin polymerization sites at podosomes.","method":"WIP-/- DCs, immunofluorescence, live imaging, calpain inhibitor experiments","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with specific molecular mechanism (calpain regulation of WASP), multiple readouts, two orthogonal approaches","pmids":["17141616"],"is_preprint":false},{"year":2006,"finding":"WIP, WASp, actin, and myosin IIA form a multiprotein complex (~1.3 MDa) in activated NK cells. Inhibitory KIR signaling decreases actin and myosin IIA recruitment to the constitutive WIP–WASp complex. PKCθ-mediated phosphorylation of WIP correlates with increased complex formation. WIP knockdown inhibits NK cell cytotoxicity.","method":"Co-immunoprecipitation/gel filtration, RNAi, kinase inhibitor studies in YTS NK cell line","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — multi-protein complex identified by co-IP/fractionation plus functional RNAi readout, single lab multiple methods","pmids":["16606694"],"is_preprint":false},{"year":2007,"finding":"WIP acts as a chaperone for WASP: WASP protein (but not mRNA) levels are severely reduced in T cells from WIP-/- mice and restored by WIP re-introduction. The WIP WASP-binding domain protects WASP from calpain-mediated degradation in vitro. Proteasome inhibitors increase WASP levels in WIP-deficient cells, indicating WASP is degraded by both calpain and the proteasome when unbound from WIP.","method":"WIP-/- mouse T cells, immunoblotting for WASP, in vitro calpain degradation assay, proteasome inhibitor treatment (MG132, bortezomib), calpain inhibitor (calpeptin)","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro degradation assay plus genetic KO plus pharmacological rescue, multiple orthogonal methods, replicated by multiple labs","pmids":["17213309"],"is_preprint":false},{"year":2007,"finding":"Drosophila D-WIP (WIP ortholog) is expressed specifically in myoblasts and bridges the WASp-Arp2/3 actin nucleation system to the myoblast adhesion molecules Dumbfounded and Sticks and Stones, recruiting the actin polymerization machinery to fusion sites. Loss of D-WIP or Wsp blocks myoblast fusion at the stage of fusion pore enlargement.","method":"Drosophila genetic analysis, immunoprecipitation, immunofluorescence in embryos","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis in Drosophila ortholog plus biochemical interaction, defined cellular phenotype","pmids":["17419994"],"is_preprint":false},{"year":2007,"finding":"WASP expression requires WIP: WASP gene transfer yields high WASP expression only when WIP is co-expressed in K562 cells; WIP knockdown in T cells reduces WASP levels. The minimal WIP region that rescues WASP expression is the WASP-binding domain, but this minimal domain is insufficient to rescue WASP-dependent NFAT-mediated IL-2 transcription.","method":"Co-expression in K562 cells, siRNA knockdown in T cells, domain-mapping WIP mutants, reporter assay","journal":"International immunology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — RNAi plus re-expression plus domain mapping, single lab, multiple orthogonal approaches","pmids":["17205972"],"is_preprint":false},{"year":2007,"finding":"WIP–WASP complex mediates TCR-induced NFAT activation without dissociation: PKCθ-mediated phosphorylation of WIP Ser488 does not cause WIP–WASP dissociation; WIP–WASP complexes persist after TCR stimulation; a WIP–WASP fusion protein efficiently mediates NFAT activation. The WIP N-terminus (polyproline and WH2 domain) is inhibitory for TCR-mediated NFAT activation.","method":"Co-IP after TCR stimulation, WIP-WASP fusion protein, domain truncation, NFAT reporter assay in Jurkat cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple WIP mutants and fusion constructs, co-IP, reporter assay, single lab","pmids":["17711847"],"is_preprint":false},{"year":2008,"finding":"WIP is indispensable for NK cell cytotoxicity: WIP knockdown completely inhibits cytolysis; WIP overexpression enhances cytolytic ability. WIP co-localizes with lytic granules and segregates to the lysosomal fraction. WIP knockdown inhibits polarization of lytic granules to the immune synapse (but not conjugate formation), and granule-WIP interaction is independent of WASp.","method":"RNAi knockdown and overexpression in YTS NK cells, cytotoxicity assays, subcellular fractionation, immunofluorescence co-localization","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Moderate — RNAi plus overexpression plus fractionation, multiple orthogonal methods, single lab","pmids":["18258743"],"is_preprint":false},{"year":2008,"finding":"EFC/F-BAR proteins (FBP17 and Toca-1) activate N-WASP–WIP complex-mediated actin polymerization in a membrane curvature-dependent manner, requiring phosphatidylserine-containing membranes and Toca-1/FBP17 but not Cdc42 or PIP2. Toca-1/FBP17 recruit N-WASP–WIP to the membrane via conserved acidic residues near their SH3 domains.","method":"In vitro actin polymerization assays with liposomes of defined curvature, domain mutants","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution with defined membrane curvature plus domain mapping, single lab","pmids":["18923421"],"is_preprint":false},{"year":2009,"finding":"WIP is essential for IL-2 signaling in T cells: WIP/WASP double-KO (DKO) T cells (unlike WASP-KO alone) fail to respond to IL-2, as evidenced by failure to up-regulate CD25, phosphorylate STAT5, or induce STAT5-dependent genes after antigen stimulation. DKO T cells have a disrupted subcortical actin cytoskeleton and impaired TCR-triggered actin polymerization.","method":"WIP/WASP double-KO mouse model, IL-2 signaling assays (STAT5 phosphorylation, CD25 up-regulation), F-actin content measurements","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean double-KO genetic model with multiple signaling readouts compared to single KO controls","pmids":["19359486"],"is_preprint":false},{"year":2010,"finding":"The cortactin-binding domain of WIP (residues 110–170) is essential for podosome formation and MMP-mediated extracellular matrix degradation by dendritic cells; WIP-/- DCs can synthesize MMPs but fail to degrade matrix. Lentiviral rescue with WIPΔ110–170 restores disorganized podosomes but not matrix degradation.","method":"WIP-/- DCs, lentiviral rescue with domain deletion mutant, gelatin degradation assay, immunofluorescence for MMP localization","journal":"European journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with domain-specific rescue, functional matrix degradation readout, single lab","pmids":["20952093"],"is_preprint":false},{"year":2011,"finding":"Blown fuse (Blow), an FCM-specific Drosophila protein, modulates the stability of the WASP–WIP complex by competing with WASP for WIP binding; this competition drives rapid exchange of WASP, WIP, and G-actin within the podosome-like structure, which is required for fusion pore formation in myoblast fusion.","method":"Drosophila genetics, biochemical competition assays, co-IP, FRAP","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic and biochemical competition experiments, defined cellular process, Drosophila ortholog","pmids":["21571220"],"is_preprint":false},{"year":2012,"finding":"WIP deficiency caused by a homozygous WIPF1 stop codon mutation (c.1301C>G) results in undetectable WASP protein (despite normal WAS mRNA), establishing that WIP stabilizes WASP in human T cells. Introduction of WIP into patient T cells restored WASP expression.","method":"Patient cells with WIPF1 mutation, immunoblotting for WASP, lentiviral WIP re-introduction","journal":"The Journal of experimental medicine","confidence":"High","confidence_rationale":"Tier 2 / Strong — natural human genetic experiment with molecular rescue, replicated mechanistic concept established in mice","pmids":["22231303"],"is_preprint":false},{"year":2013,"finding":"WIP (or its homolog WIRE) is an essential link between Nck and N-WASP for Arp2/3-dependent actin assembly: N-WASP recruitment to vaccinia virus depends on WIP (not on direct Nck–N-WASP interaction). WIP contains two Nck-binding sites and is recruited to virus by the second SH3 domain of Nck while bound to N-WASP. The first and third SH3 domains of Nck are required to stimulate actin assembly but not to recruit the WIP:N-WASP complex.","method":"MEFs lacking Nck, WIP, or N-WASP; vaccinia actin-tail assay; domain mutants of Nck and WIP; co-IP","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Strong — triple-KO MEF analysis plus domain mapping plus functional actin assembly assay, multiple orthogonal approaches","pmids":["23707428"],"is_preprint":false},{"year":2014,"finding":"WIP and WASp form two distinct molecular interfaces in cells: (i) WH1 domain of WASp with C-terminal WIP, dependent on PKCθ-mediated WIP phosphorylation (in response to TCR activation); (ii) VCA domain of WASp with N-terminal WIP, dependent on actin (inhibited by latrunculin A). WASp activation involves dissociation of interface (i) while interface (ii) remains, exposing the WASp ubiquitylation site and promoting degradation.","method":"Triple-color FRET (3FRET) in live T cells, PKCθ inhibitors, latrunculin A, phosphomimetic WIP mutants","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — novel 3FRET technology with pharmacological validations and mutant controls, single lab multiple orthogonal methods","pmids":["24962707"],"is_preprint":false},{"year":2014,"finding":"Tyrosine phosphorylation of WIP (mediated by Bruton's tyrosine kinase, Btk) releases bound WASP from the WIP–WASP complex; in the absence of WIP–WASP binding, WASP is rapidly degraded. WIP phosphomimics abolish WIP–WASP interaction and disrupt podosomes; WIP lacking tyrosine phosphorylation extends podosome lifetimes. Btk was identified as a kinase regulating WIP tyrosine phosphorylation.","method":"WIP knockdown, phosphomimic/phosphonull WIP mutants, kinase screen with inhibitors, podosome assay, matrix degradation assay in macrophages","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — phosphomimetic domain mutants, kinase identification, functional podosome/degradation readouts, single lab multiple methods","pmids":["25413351"],"is_preprint":false},{"year":2014,"finding":"WIP binding to actin (via its actin-binding domain, ABD), independently of its binding to WASp, is critical for the integrity of the actin cytoskeleton in T cells and for their migration; WIPΔABD mice have T cells with normal WASp levels but decreased F-actin, disorganized actin cytoskeleton, impaired chemotaxis, and defective homing to lymph nodes.","method":"Knock-in mice expressing WIP lacking the ABD (WIPΔABD), F-actin staining, chemotaxis assays, lymph node homing assays","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean knock-in mouse model isolating actin-binding from WASp-binding, multiple functional readouts","pmids":["25246631"],"is_preprint":false},{"year":2014,"finding":"WIP binding to F-actin (via ABD) is required for focal adhesion assembly and stress fiber formation in fibroblasts. WIP-/- fibroblasts have defective focal adhesions, increased G-actin levels, and reduced nuclear MRTF-A/SRF activity; constitutively nuclear MRTF-A or active SRF restores these defects, establishing a WIP–actin–MRTF–SRF axis in cell adhesion.","method":"WIP-/- fibroblasts, knock-in WIP mutant (fails to bind actin), MRTF-A nuclear translocation, SRF reporter, focal adhesion immunostaining","journal":"Molecular and cellular biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO plus knock-in domain mutant plus downstream transcription factor rescue, multiple orthogonal methods","pmids":["24797074"],"is_preprint":false},{"year":2014,"finding":"NMR structural characterization showed WIP N-terminal ABM is intrinsically disordered but has residual helical (residues 30–42) and β-strand (residues 44–62) propensities that echo the actin-bound conformation; residues 17–25 preceding the canonical ABM also show β-strand propensity, suggesting the WIP–actin interaction epitope extends to the N-terminal polyproline region.","method":"NMR (protonless 13C'-detected spectroscopy), secondary chemical shifts, RDC measurements","journal":"The FEBS journal","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — structural NMR data in isolated domain, no functional mutagenesis validation, single lab","pmids":["25495558"],"is_preprint":false},{"year":2015,"finding":"Intersectin adaptor proteins ITSN1 and ITSN2 interact with WIP via SH3 domain–proline-rich motif interactions (middle part of WIP proline-rich region). ITSN1, WIP, and N-WASP form a trimeric complex in cells. Endogenous ITSN1 co-localizes with WIP at invadopodia in breast cancer cells.","method":"Co-immunoprecipitation, GST-SH3 pulldown, immunofluorescence co-localization in MDA-MB-231 cells","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP plus domain mapping plus co-localization, single lab","pmids":["25797047"],"is_preprint":false},{"year":2016,"finding":"DOCK8 is connected to WASp and actin in T cells through WIP acting as a bridge: WIP co-immunoprecipitates with DOCK8 and WASp. DOCK8 guanine nucleotide exchange factor activity is essential for WASp activation, F-actin assembly, immune synapse/actin foci formation, mechanotransduction, T cell transendothelial migration, and lymph node homing—all of which also depend on WASp, placing DOCK8 and WASp in the same actin-regulatory pathway via WIP.","method":"Co-immunoprecipitation, T cell functional assays (immune synapse, foci, migration), T cells from DOCK8-deficient and WAS patients","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP establishing DOCK8–WIP–WASp complex plus multiple functional assays in human and mouse primary cells","pmids":["27599296"],"is_preprint":false},{"year":2016,"finding":"WIP and WICH/WIRE play non-redundant roles in invadopodium formation in breast cancer cells: WIP interacts with N-WASP and cortactin and is essential for invadopodium assembly, while WICH/WIRE regulates N-WASP activation to control invadopodium maturation and degradative activity. Nck interaction with WIP modulates invadopodium maturation.","method":"RNAi knockdown of WIP and WICH/WIRE, co-immunoprecipitation, invadopodium formation/matrix degradation assays, TIRF microscopy","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — RNAi with rescue, co-IP, functional invadopodium assay, multiple orthogonal methods","pmids":["27009365"],"is_preprint":false},{"year":2016,"finding":"WIP controls tumor growth by stabilizing the YAP/TAZ complex via the endocytic/endosomal system: when WIP levels are high, the β-catenin destruction complex (APC–axin–GSK3) is sequestered to multi-vesicular body compartments, inhibiting YAP/TAZ degradation. YAP/TAZ stability is dependent on Rac, PAK, and mDia, and is Hippo-independent.","method":"WIP knockdown/overexpression in cancer cells, subcellular fractionation, co-IP for destruction complex, Rac/PAK/mDia inhibitors, in vivo xenograft","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — fractionation plus co-IP plus pharmacological inhibitors plus in vivo, single lab","pmids":["27851961"],"is_preprint":false},{"year":2017,"finding":"Mutant p53 oncogenic activity is driven by WIP: WIP is phosphorylated by AKT2 downstream of mtp53/p63-enhanced PI3K/AKT2-mediated integrin/receptor recycling pathways. WIP regulates YAP/TAZ stability; WIP knockdown reduces CSC markers (CD133, CD44, YAP/TAZ) and tumor growth in vivo.","method":"WIP knockdown/overexpression, mtp53 overexpression, co-IP for AKT2-WIP interaction, in vivo tumor growth assays","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP showing AKT2–WIP interaction plus phosphorylation plus functional knockdown with in vivo validation, single lab","pmids":["28166194"],"is_preprint":false},{"year":2017,"finding":"NMR and FRET analysis of the WIP C-terminal (residues 442–492)–WASp (residues 20–158) complex revealed a pleckstrin homology-like domain with mixed α/β fold; WIP residues 454–456 are the major contributor to WASp affinity, and residues 449–451 have the largest effect on WASp ubiquitylation and degradation. WIP binding to WASp is inversely linked to WASp ubiquitylation.","method":"NMR structure of complex, FRET in vivo, biochemical ubiquitylation assays, WIP peptide mutants","journal":"ACS chemical biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure plus FRET plus ubiquitylation assay, multiple orthogonal methods, single lab","pmids":["29215267"],"is_preprint":false},{"year":2017,"finding":"At yeast endocytic sites, WASP and WIP accumulate to a threshold level through multivalent SH3 domain–PRM interactions involving linker proteins; Arp2/3-mediated actin assembly initiation is tightly coupled to reaching threshold levels of WASP and WIP (not to recruitment kinetics or autoinhibition release), giving actin assembly onset switch-like behavior.","method":"Quantitative live-cell fluorescence imaging of endocytic sites, yeast genetics (SH3/PRM mutants)","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 / Moderate — quantitative imaging with genetic perturbations in yeast ortholog system, single lab","pmids":["28813247"],"is_preprint":false},{"year":2018,"finding":"WIP interacts with RhoA: in lung adenocarcinoma cells, WIP knockdown reduces RhoA levels and WIP co-immunoprecipitates with RhoA; WIP regulates invasion, EMT, and anchorage-independent growth via RhoA.","method":"Co-immunoprecipitation, siRNA knockdown, invasion assays, RhoA immunoblotting","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single co-IP and knockdown, single lab, cancer cell line only","pmids":["27939884"],"is_preprint":false},{"year":2018,"finding":"WIP/ITSN1 complex co-localizes with RAB4-positive fast-recycling endosomes and is involved in transferrin receptor recycling. ITSN1 recruits WIP to RAB4-positive vesicles. WIP enhances N-WASP–ITSN1 interaction and ITSN1/β-actin association, and the WIP/ITSN1-L complex promotes filopodia-like protrusion formation.","method":"Co-immunoprecipitation, transferrin recycling assay, co-localization with Rab4, overexpression in MCF-7 cells","journal":"Gene","confidence":"Low","confidence_rationale":"Tier 3 / Weak — co-IP and co-localization, single lab, no mechanistic dissection of which WIP domain mediates vesicle function","pmids":["29958948"],"is_preprint":false},{"year":2019,"finding":"In C. elegans intestine, WIP-1 promotes scission of clathrin-coated pits by directly binding G-actin (independent of WSP-1/WASP); the cortactin-binding domain of WIP-1 serves as the binding interface for DBN-1 (Abp1), and DBN-1–F-actin interaction is essential for Dynamin-1 (DYN-1) recruitment at endocytic sites.","method":"RNAi knockdown, live imaging of CCP scission in C. elegans intestine, domain mapping for WIP-1–DBN-1 interaction","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — C. elegans ortholog, RNAi, live imaging, domain-specific interaction, WASP-independent mechanism established","pmids":["31118234"],"is_preprint":false},{"year":2020,"finding":"WIP depletion increases reactive oxygen species and reduces NRF2 levels in glioblastoma cells. WIP stabilizes NRF2 by restraining KEAP1 E3 ligase activity; increased KEAP1 activity in WIP-depleted cells depends on actin cytoskeleton organization (via KEAP1–F-actin binding), not protection of KEAP1 from autophagic degradation.","method":"WIP knockdown, KEAP1 knockdown, NRF2ΔETGE overexpression, ROS measurements, co-IP of KEAP1–F-actin","journal":"Antioxidants (Basel, Switzerland)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic rescue experiments plus co-IP identifying KEAP1–actin connection, single lab","pmids":["32825452"],"is_preprint":false},{"year":2023,"finding":"WIPF1 interacts with ACTN4 to regulate podosome formation, matrix degradation, and actin polymerization in extravillous trophoblasts (EVTs); the ARG54 site of WIPF1 is implicated in this interaction. WIPF1 knockdown impairs EVT cell migration and trophoblast differentiation; WIPF1 is downregulated in RSA patient EVTs.","method":"Co-immunoprecipitation, WIPF1 knockdown in hTSC-derived EVTs, podosome/matrix degradation assays, site-directed mutagenesis (ARG54)","journal":"Genes & diseases","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with mutagenesis plus functional podosome/migration assay, single lab","pmids":["40821124"],"is_preprint":false},{"year":2023,"finding":"WIPF1 promotes gastric cancer cell proliferation, invasion, and migration in a myocardin (MYOCD)-dependent manner by activating the PI3K/AKT signaling pathway; MYOCD transactivates WIPF1 transcription and silencing WIPF1 significantly represses PI3K/AKT activation.","method":"siRNA knockdown, overexpression, in vitro and xenograft assays, PI3K/AKT pathway immunoblotting, MYOCD rescue experiments","journal":"iScience","confidence":"Low","confidence_rationale":"Tier 3 / Weak — knockdown/overexpression with pathway readout, single lab, mechanism not dissected at molecular level","pmids":["38026208"],"is_preprint":false},{"year":2005,"finding":"C. elegans WIP-1 physically interacts with WSP-1 (WASP/N-WASP homolog) by yeast two-hybrid. RNAi knockdown of wip-1 decreases WSP-1 protein levels (not mRNA), and wsp-1 RNAi decreases WIP-1 protein levels (not mRNA), establishing mutual protein stabilization. WIP-1 RNAi causes embryonic lethality with hypodermal cell migration defects (ventral enclosure) similar to wsp-1 RNAi.","method":"Yeast two-hybrid, RNAi, Western blot, immunostaining in C. elegans embryos","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — ortholog functional studies with yeast two-hybrid, RNAi, protein level quantitation, and defined morphogenetic phenotype","pmids":["16378591"],"is_preprint":false},{"year":2011,"finding":"WIP-deficient hippocampal neurons show enlarged somas, overgrowth of neuritic and dendritic branches at early developmental stages, increased dendritic arborization, and increased amplitude and frequency of miniature excitatory postsynaptic currents, identifying WIP as a negative regulator of neuronal maturation and synaptic activity.","method":"WIP-/- mouse hippocampal neurons, morphometric analysis, electrophysiology (mEPSC recording)","journal":"Cerebral cortex (New York, N.Y. : 1991)","confidence":"High","confidence_rationale":"Tier 2 / Moderate — genetic KO with quantitative morphology and electrophysiology, two orthogonal readouts, single lab","pmids":["21810783"],"is_preprint":false},{"year":2014,"finding":"WIP absence in dendritic spines increases spine size and F-actin levels through a RhoA/ROCK/profilinIIa-dependent (N-WASP/Arp2/3-independent) mechanism. WIP deficiency causes transcriptional upregulation of neutral sphingomyelinase (NSM) via active RhoA, reducing membrane sphingomyelin, which in turn enhances RhoA membrane association and raft partitioning. NSM inhibition or sphingomyelin addition reverses RhoA, F-actin, and functional anomalies in WIP-/- synapses.","method":"WIP-/- mouse neurons, pharmacological inhibition of NSM, sphingomyelin supplementation, RhoA localization assay, F-actin staining, Arp2/3 co-IP","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO plus pharmacological rescue plus lipid supplementation, single lab, novel mechanism","pmids":["24698977"],"is_preprint":false}],"current_model":"WIPF1 (WIP) is an intrinsically disordered, multifunctional scaffold protein that binds WASP/N-WASP (through its C-terminal WASp-binding domain), actin (through its N-terminal WH2/ABD domain), profilin, Nck, cortactin, DOCK8, and other SH3-containing adaptors, thereby acting as a core hub that (i) stabilizes WASP/N-WASP from calpain- and proteasome-mediated degradation, (ii) recruits and positions the N-WASP–Arp2/3 actin nucleation machinery to sites of actin polymerization (immune synapses, podosomes, invadopodia, filopodia, endocytic pits, pathogen tails), (iii) directly stabilizes F-actin filaments, (iv) integrates upstream signals (Cdc42 via Toca-1/FBP17, Nck, PKCθ phosphorylation, Btk-mediated tyrosine phosphorylation, AKT2) to regulate when and where WASP is activated or degraded, and (v) exerts WASP-independent functions including lytic granule polarization in NK cells, MRTF-SRF-driven focal adhesion assembly, YAP/TAZ stabilization through endosomal sequestration of the destruction complex, and regulation of NRF2 oxidative-stress response through actin-dependent KEAP1 restraint."},"narrative":{"mechanistic_narrative":"WIPF1 (WIP) is an intrinsically disordered scaffold protein that couples actin nucleation machinery to upstream signals and controls the stability and localization of WASP/N-WASP at sites of actin polymerization [PMID:9405671, PMID:11331876]. It was first identified through its direct interaction with WASP, binding at a site distinct from the Cdc42-binding region while carrying its own N-terminal actin- and profilin-binding motifs required to drive actin polymerization [PMID:9405671]; the structural basis of the C-terminal interaction was resolved as a peptide that wraps around the N-WASP EVH1/WH1 domain, explaining how WAS disease mutations disrupt WIP binding [PMID:12437929, PMID:29215267]. A central function of WIP is to act as a chaperone that protects WASP from calpain- and proteasome-mediated degradation: WASP protein but not mRNA is lost in WIP-deficient cells and restored by re-expression, a mechanism conserved from C. elegans through mouse to a human WIPF1 stop-codon patient in whom WASP was undetectable [PMID:17213309, PMID:22231303, PMID:16378591]. WIP positions and activates the N-WASP–Arp2/3 system at diverse actin structures, integrating signals from Nck, cortactin, the F-BAR proteins Toca-1/FBP17 in a membrane-curvature-dependent manner, and DOCK8 [PMID:9694849, PMID:12620186, PMID:15260990, PMID:18923421, PMID:23707428, PMID:27599296]. Through these activities WIP drives filopodia, membrane ruffling, pathogen actin tails, podosomes and invadopodia with associated matrix degradation, and clathrin-mediated endocytosis [PMID:10878810, PMID:11331876, PMID:12724353, PMID:17141616, PMID:20952093, PMID:27009365, PMID:28813247]. In the immune system WIP is essential for T-cell activation, immune-synapse formation, and IL-2 signaling, for FcεRI-driven mast cell signaling via stabilization of Syk, and for NK-cell cytotoxicity through WASP-independent polarization of lytic granules [PMID:11869681, PMID:14757742, PMID:18258743, PMID:19359486]. WIP also exerts WASP-independent functions through direct actin binding, supporting cytoskeletal integrity and T-cell migration, focal adhesion assembly via an actin–MRTF–SRF axis, and regulation of neuronal maturation and synaptic activity [PMID:25246631, PMID:24797074, PMID:21810783]. Loss-of-function WIPF1 mutation in humans causes a WAS-like primary immunodeficiency with absent WASP [PMID:22231303].","teleology":[{"year":1997,"claim":"Established WIP as a physical and functional partner of WASP, defining its core domain architecture (actin/profilin-binding N-terminus, distinct WASP-binding site) and its capacity to drive actin polymerization.","evidence":"Yeast two-hybrid, co-IP from lymphocytes, and truncation-mutant expression in B cells","pmids":["9405671"],"confidence":"High","gaps":["Did not establish whether WIP regulates WASP stability or only localization","Mechanism of actin polymerization induction not resolved biochemically"]},{"year":1998,"claim":"Showed WIP links the adaptor Nck and profilin to the cytoskeleton through domain-specific interactions, placing WIP within receptor-proximal signaling adaptor networks.","evidence":"GST-Nck pulldown and WIP fragment domain mapping in B-cell lysates","pmids":["9694849"],"confidence":"Medium","gaps":["Single lab, no functional readout for the Nck–WIP interaction","Did not test reciprocity in vivo"]},{"year":1999,"claim":"Demonstrated functional conservation of the WIP–WASP partnership in endocytosis and cell polarity, mapping the requirement to the WH2 actin- and profilin-binding domains.","evidence":"Yeast genetic suppression/complementation with verprolin orthologs and WIP domain mutants","pmids":["9742397","10358064"],"confidence":"High","gaps":["Yeast system may not capture mammalian-specific regulatory inputs","Did not address WASP stabilization role"]},{"year":2000,"claim":"Defined WIP as the recruiter of N-WASP to actin-polymerization sites via the N-WASP WH1 domain, integrating Cdc42 and SH2/SH3-adaptor pathways during pathogen actin-based motility.","evidence":"Cell-based vaccinia and Shigella actin-tail assays with domain mutants and dominant negatives","pmids":["10878810"],"confidence":"High","gaps":["Did not resolve how upstream signals choose between recruitment and activation","Quantitative contribution of WIP vs other recruiters unclear"]},{"year":2001,"claim":"Established the biochemical activity of WIP: direct N-WASP and actin binding, F-actin stabilization, and a required role with N-WASP in filopodium formation.","evidence":"In vitro pyrene-actin assays, microinjection with function-blocking antibodies, and co-IP/imaging","pmids":["11331876","11687573"],"confidence":"High","gaps":["Apparent inhibition of Arp2/3 nucleation in vitro vs activation in cells not reconciled","Regulation of the WIP–N-WASP unit by phosphorylation not yet addressed"]},{"year":2002,"claim":"Genetic loss-of-function established WIP as essential for T-cell activation and immune synapse formation, with cell-type-specific (T vs B) consequences, and showed WIP supports N-WASP recruitment at vesicle surfaces.","evidence":"WIP-knockout mouse T/B cell assays and N-WASP-deficient cell reconstitution with domain mutants","pmids":["11869681","12147689"],"confidence":"High","gaps":["Did not distinguish actin-scaffolding from WASP-stabilizing contributions to the T-cell phenotype","Molecular basis of the divergent B-cell phenotype unresolved"]},{"year":2002,"claim":"Resolved the structural recognition mechanism between WIP and the N-WASP EVH1/WH1 domain, providing a molecular explanation for WAS-associated missense mutations that disrupt WIP binding.","evidence":"NMR structure of the N-WASP WH1 domain bound to a WIP peptide","pmids":["12437929"],"confidence":"High","gaps":["Structure of full-length disordered WIP not determined","Did not address the actin-dependent second interface"]},{"year":2003,"claim":"Extended WIP's effector network to cortactin and to Rac1/PDGF-driven membrane dynamics, with the actin-binding domain required for ruffle formation.","evidence":"Cortactin SH3 pulldowns and in vitro Arp2/3 assays; anti-WIP microinjection and domain mutants in fibroblasts","pmids":["12620186","12724353"],"confidence":"High","gaps":["How cortactin and N-WASP inputs are coordinated on WIP not resolved","In vivo relevance of cortactin synergy to specific actin structures untested"]},{"year":2004,"claim":"Identified F-BAR protein Toca-1 as an activator of the predominant cellular N-WASP–WIP complex, establishing cooperative Cdc42 effector logic, and broadened WIP's signaling roles to mast cell FcεRI signaling through Syk stabilization.","evidence":"Biochemical purification and in vitro actin assembly; WIP-/- mast cells with Syk co-IP and stability assays","pmids":["15260990","14757742"],"confidence":"High","gaps":["Whether Syk stabilization is mechanistically analogous to WASP chaperoning untested","Toca-1 vs FBP17 specificity not yet defined"]},{"year":2006,"claim":"Established WIP as essential for podosome formation and revealed its role in regulating calpain-mediated WASP cleavage, and identified a WASP-containing actin/myosin megacomplex in NK cells modulated by PKCθ phosphorylation.","evidence":"WIP-/- dendritic cells with calpain inhibitors; co-IP/gel filtration and RNAi in NK cell lines","pmids":["17141616","16606694"],"confidence":"High","gaps":["How phosphorylation controls complex composition not mechanistically resolved","Link between podosome defect and WASP degradation only correlative"]},{"year":2007,"claim":"Defined WIP's chaperone function in molecular terms—the WASP-binding domain protects WASP from calpain and proteasomal degradation—while showing this minimal protection is insufficient for full WASP-dependent signaling, and extended conserved WIP–WASP partnership to Drosophila myoblast fusion.","evidence":"WIP-/- T cells with in vitro calpain assays and proteasome/calpain inhibitors; K562 co-expression and domain mapping; Drosophila genetics","pmids":["17213309","17205972","17419994","17711847"],"confidence":"High","gaps":["Identity of the proteasomal E3 ligase for free WASP not defined","How the WIP N-terminus inhibits NFAT activation mechanistically unclear"]},{"year":2008,"claim":"Established a WASP-independent role for WIP in NK-cell cytotoxicity through lytic granule polarization, demonstrating WIP functions beyond the WIP–WASP axis.","evidence":"RNAi/overexpression in NK cell lines with cytotoxicity assays, subcellular fractionation, and co-localization with lytic granules","pmids":["18258743","18923421"],"confidence":"High","gaps":["Molecular tether linking WIP to lytic granules unidentified","Membrane-curvature activation studied in vitro only"]},{"year":2009,"claim":"Used a WIP/WASP double knockout to show WIP is required for IL-2/STAT5 signaling and subcortical actin integrity beyond WASP loss alone, separating WIP-specific from WASP-shared functions.","evidence":"WIP/WASP double-KO mouse T cells with STAT5 phosphorylation, CD25, and F-actin readouts versus single-KO controls","pmids":["19359486"],"confidence":"High","gaps":["Direct molecular link between WIP and the IL-2 receptor signaling apparatus not defined","Whether actin defect causes the signaling defect untested"]},{"year":2010,"claim":"Mapped the cortactin-binding region of WIP as essential for matrix-degrading function of podosomes, dissociating podosome assembly from degradative activity.","evidence":"WIP-/- dendritic cells rescued with a domain-deletion mutant and gelatin degradation assays","pmids":["20952093"],"confidence":"High","gaps":["How the cortactin-binding region directs MMP delivery not resolved","Direct cortactin requirement not genetically isolated"]},{"year":2011,"claim":"Revealed dynamic competitive regulation of the WASP–WIP complex (by Blow in Drosophila) driving rapid component exchange required for fusion-pore formation, and identified WIP as a negative regulator of neuronal maturation and synaptic activity.","evidence":"Drosophila genetics with competition and FRAP assays; WIP-/- mouse hippocampal neuron morphometry and electrophysiology","pmids":["21571220","21810783"],"confidence":"High","gaps":["Whether mammalian factors analogous to Blow regulate WASP–WIP exchange unknown","Molecular pathway underlying the neuronal phenotype not yet defined here"]},{"year":2012,"claim":"Confirmed in humans that WIPF1 loss-of-function abolishes WASP protein and causes a WAS-like immunodeficiency, validating the chaperone model as clinically relevant.","evidence":"Patient cells with a homozygous WIPF1 stop-codon mutation, WASP immunoblotting, and lentiviral WIP rescue","pmids":["22231303"],"confidence":"High","gaps":["Genotype-phenotype range of WIPF1 deficiency not delineated","Did not address WASP-independent contributions to patient phenotype"]},{"year":2013,"claim":"Established WIP (or WIRE) as the obligatory link between Nck and N-WASP for Arp2/3-dependent actin assembly, separating the recruitment step from the actin-stimulation step within Nck.","evidence":"Nck-, WIP-, and N-WASP-deficient MEFs in vaccinia actin-tail assays with Nck/WIP domain mutants","pmids":["23707428"],"confidence":"High","gaps":["How the first/third Nck SH3 domains stimulate assembly mechanistically unresolved","Generality beyond pathogen tails to physiological structures not tested"]},{"year":2014,"claim":"Defined two distinct WIP–WASP interfaces (a phosphorylation-dependent WH1 interface and an actin-dependent VCA interface) and showed WASP activation couples interface remodeling to exposure of its ubiquitylation site, mechanistically linking activation to degradation; also isolated WIP's WASP-independent actin-binding functions in T-cell migration and focal-adhesion/MRTF–SRF signaling, and resolved the disordered actin-binding motif by NMR.","evidence":"Triple-color FRET in live T cells with PKCθ inhibitors/phosphomimetics; Btk kinase identification with phosphomutants; WIPΔABD knock-in mice; WIP-/- fibroblasts with MRTF/SRF rescue; NMR of the ABM","pmids":["24962707","25413351","25246631","24797074","25495558"],"confidence":"High","gaps":["How phospho- and actin-dependent interfaces are temporally coordinated in vivo not fully resolved","Identity of the WASP ubiquitin ligase still unaddressed"]},{"year":2015,"claim":"Extended the WIP scaffold to intersectin adaptors, forming an ITSN1–WIP–N-WASP trimer that localizes to invadopodia.","evidence":"Co-IP, GST-SH3 pulldown, and co-localization in breast cancer cells","pmids":["25797047"],"confidence":"Medium","gaps":["Functional requirement of ITSN1 for invadopodia not tested here","Single lab, no reciprocal in vivo validation"]},{"year":2016,"claim":"Bridged DOCK8 to WASP-dependent actin regulation via WIP in T cells, distinguished non-redundant WIP versus WIRE roles in invadopodium assembly versus maturation, and uncovered a WASP-independent oncogenic role for WIP in stabilizing YAP/TAZ via endosomal sequestration of the β-catenin destruction complex.","evidence":"Co-IP and functional T-cell assays in DOCK8/WAS patient cells; RNAi with invadopodium assays; WIP knockdown/overexpression with fractionation, inhibitors, and xenografts","pmids":["27599296","27009365","27851961"],"confidence":"High","gaps":["Mechanism of endosomal sequestration of the destruction complex not resolved at molecular detail","How DOCK8 GEF activity is coupled to the WIP scaffold unclear"]},{"year":2017,"claim":"Provided high-resolution structure of the WIP C-terminus–WASp complex linking specific residues to WASp affinity and ubiquitylation, and placed WIP downstream of AKT2 in mutant-p53-driven tumorigenesis controlling YAP/TAZ and cancer stem-cell markers.","evidence":"NMR/FRET of the WIP–WASp complex with peptide mutants and ubiquitylation assays; co-IP, phosphorylation, and in vivo tumor assays","pmids":["29215267","28166194"],"confidence":"High","gaps":["Structural data did not capture the full intact complex in cells","AKT2-WIP phosphosite functional consequences from single lab"]},{"year":2018,"claim":"Implicated WIP in RAB4-positive recycling endosome trafficking with ITSN1 and in RhoA-dependent cancer cell invasion, broadening its membrane-trafficking and tumor-promoting roles.","evidence":"Co-IP, transferrin recycling and co-localization assays in MCF-7; co-IP and invasion assays in lung adenocarcinoma","pmids":["29958948","27939884"],"confidence":"Low","gaps":["Co-IP/co-localization only without domain dissection of vesicle function","RhoA interaction shown by single co-IP without reciprocal validation"]},{"year":2019,"claim":"Demonstrated a WASP-independent endocytic function of WIP-1 in scission of clathrin-coated pits via direct G-actin binding and cortactin-domain-mediated recruitment of DBN-1/Abp1 and dynamin.","evidence":"RNAi and live imaging in C. elegans intestine with domain mapping of the WIP-1–DBN-1 interaction","pmids":["31118234"],"confidence":"Medium","gaps":["Conservation of this WASP-independent scission role in mammals untested","How G-actin binding triggers scission mechanistically unresolved"]},{"year":2020,"claim":"Connected WIP to the NRF2 oxidative-stress response, showing WIP restrains KEAP1 E3 activity through actin-cytoskeleton-dependent KEAP1–F-actin binding rather than autophagic protection.","evidence":"WIP/KEAP1 knockdown, NRF2 mutant overexpression, ROS measurement, and KEAP1–F-actin co-IP in glioblastoma cells","pmids":["32825452"],"confidence":"Medium","gaps":["Single lab, no in vivo confirmation","Direct mechanism by which actin organization controls KEAP1 activity unresolved"]},{"year":2023,"claim":"Extended WIPF1 actin functions to extravillous trophoblast podosomes via ACTN4 interaction and to MYOCD-driven PI3K/AKT signaling in gastric cancer, implicating WIPF1 in trophoblast differentiation/disease and tumor progression.","evidence":"Co-IP with ARG54 site mutagenesis and migration/podosome assays in trophoblasts; knockdown/overexpression with pathway readouts and MYOCD rescue in gastric cancer","pmids":["40821124","38026208"],"confidence":"Medium","gaps":["ACTN4 interaction from single lab without reciprocal in vivo validation","Gastric cancer mechanism not dissected at molecular level"]},{"year":null,"claim":"How the multiple WIP interfaces, phosphorylation inputs (PKCθ, Btk, AKT2), and partner choices are integrated in space and time to switch between WASP stabilization, WASP activation/degradation, and WASP-independent functions remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified model coupling signal input to interface remodeling in vivo","E3 ligase mediating WASP/WIP-regulated ubiquitylation unidentified","Structural basis of full-length WIP scaffolding not determined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,5,29,30,41]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[1,26,33,32]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[16,5,12,37]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[16,25,45]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[5,6,29,30]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,6]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[20,40,41]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[11,14,21]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[8,13,20,22,33]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[2,9,38,41]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[12,26,27,30]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[25,35,36]}],"complexes":["WIP–WASP/N-WASP complex","N-WASP–WIP–Arp2/3 nucleation complex","WIP–WASp–actin–myosin IIA megacomplex (NK cells)","ITSN1–WIP–N-WASP trimeric complex"],"partners":["WASP","N-WASP","NCK","CORTACTIN","TOCA-1","DOCK8","ITSN1","PROFILIN"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O43516","full_name":"WAS/WASL-interacting protein family member 1","aliases":["Protein PRPL-2","Wiskott-Aldrich syndrome protein-interacting protein","WASP-interacting protein"],"length_aa":503,"mass_kda":51.3,"function":"Plays a role in the reorganization of the actin cytoskeleton. Contributes with NCK1 and GRB2 in the recruitment and activation of WASL. May participate in regulating the subcellular localization of WASL, resulting in the disassembly of stress fibers in favor of filopodia formation. Plays a role in the formation of cell ruffles (By similarity). Plays an important role in the intracellular motility of vaccinia virus by functioning as an adapter for recruiting WASL to vaccinia virus","subcellular_location":"Cytoplasmic vesicle; Cytoplasm, cytoskeleton; Cell projection, ruffle","url":"https://www.uniprot.org/uniprotkb/O43516/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WIPF1","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"WASL","stoichiometry":10.0}],"url":"https://opencell.sf.czbiohub.org/search/WIPF1","total_profiled":1310},"omim":[{"mim_id":"617398","title":"FK506-BINDING PROTEIN 15; FKBP15","url":"https://www.omim.org/entry/617398"},{"mim_id":"616714","title":"HMG-BOX TRANSCRIPTION FACTOR 1; HBP1","url":"https://www.omim.org/entry/616714"},{"mim_id":"614493","title":"WISKOTT-ALDRICH SYNDROME 2; WAS2","url":"https://www.omim.org/entry/614493"},{"mim_id":"606420","title":"ENGULFMENT AND CELL MOTILITY GENE 1; ELMO1","url":"https://www.omim.org/entry/606420"},{"mim_id":"605035","title":"WASP PROTEIN FAMILY, MEMBER 1; WASF1","url":"https://www.omim.org/entry/605035"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":107.0}],"url":"https://www.proteinatlas.org/search/WIPF1"},"hgnc":{"alias_symbol":["WIP"],"prev_symbol":["WASPIP"]},"alphafold":{"accession":"O43516","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O43516","model_url":"https://alphafold.ebi.ac.uk/files/AF-O43516-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O43516-F1-predicted_aligned_error_v6.png","plddt_mean":58.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WIPF1","jax_strain_url":"https://www.jax.org/strain/search?query=WIPF1"},"sequence":{"accession":"O43516","fasta_url":"https://rest.uniprot.org/uniprotkb/O43516.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O43516/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O43516"}},"corpus_meta":[{"pmid":"15260990","id":"PMC_15260990","title":"Toca-1 mediates Cdc42-dependent actin nucleation by activating the N-WASP-WIP complex.","date":"2004","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/15260990","citation_count":368,"is_preprint":false},{"pmid":"9405671","id":"PMC_9405671","title":"WIP, a protein associated with wiskott-aldrich syndrome protein, induces actin polymerization and redistribution in lymphoid cells.","date":"1997","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/9405671","citation_count":311,"is_preprint":false},{"pmid":"10878810","id":"PMC_10878810","title":"A complex of N-WASP and WIP integrates signalling cascades that lead to actin polymerization.","date":"2000","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/10878810","citation_count":282,"is_preprint":false},{"pmid":"11331876","id":"PMC_11331876","title":"WIP regulates N-WASP-mediated actin polymerization and filopodium formation.","date":"2001","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/11331876","citation_count":238,"is_preprint":false},{"pmid":"18923421","id":"PMC_18923421","title":"EFC/F-BAR proteins and the N-WASP-WIP complex induce membrane curvature-dependent actin polymerization.","date":"2008","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/18923421","citation_count":152,"is_preprint":false},{"pmid":"12620186","id":"PMC_12620186","title":"Cortactin interacts with WIP in regulating Arp2/3 activation and membrane protrusion.","date":"2003","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/12620186","citation_count":151,"is_preprint":false},{"pmid":"11782451","id":"PMC_11782451","title":"A. thaliana TRANSPARENT TESTA 1 is involved in seed coat development and defines the WIP subfamily of plant zinc finger proteins.","date":"2002","source":"Genes & 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N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/39329352","citation_count":4,"is_preprint":false},{"pmid":"40821124","id":"PMC_40821124","title":"Unraveling the role of the WIPF1/ACTN4 complex in podosome formation of human placental EVTs: Insights into recurrent spontaneous abortion.","date":"2025","source":"Genes & diseases","url":"https://pubmed.ncbi.nlm.nih.gov/40821124","citation_count":2,"is_preprint":false},{"pmid":"40107543","id":"PMC_40107543","title":"Camellia sinensis WIP domain protein 3 (CsWIP3), a C2H2 zinc finger protein, mediates lignin content and regulates plant growth in tea plants.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40107543","citation_count":2,"is_preprint":false},{"pmid":"32189449","id":"PMC_32189449","title":"Controlling Smad4 signaling with a Wip.","date":"2020","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/32189449","citation_count":1,"is_preprint":false},{"pmid":"40415347","id":"PMC_40415347","title":"WIPF1 regulates stemness in small cell lung cancer.","date":"2025","source":"Science progress","url":"https://pubmed.ncbi.nlm.nih.gov/40415347","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":46558,"output_tokens":12138,"usd":0.160872,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":23251,"output_tokens":7976,"usd":0.157828,"stage2_stop_reason":"end_turn"},"total_usd":0.3187,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1997,\n      \"finding\": \"WIP (WIPF1) was identified as a WASP-interacting protein via yeast two-hybrid; it coimmunoprecipitates with WASP from lymphocytes, binds WASP at a site distinct from the Cdc42 binding site, and has actin and profilin binding motifs. Expression of WIP in B cells induced actin polymerization and cerebriform projections; a WIP truncation mutant lacking the actin-binding motif failed to do so, establishing the actin-binding domain as required for this function.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, expression in human B cells with truncation mutant\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, domain mapping with truncation mutant, functional readout; foundational paper replicated across many subsequent studies\",\n      \"pmids\": [\"9405671\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"WIP binds to the adaptor protein Nck via the second SH3 domain of Nck; the Nck-binding site on WIP (amino acids 321–415) is distinct from the WASP-binding site (amino acids 416–488). Profilin is found in Nck precipitates, suggesting Nck links to the cytoskeleton via WIP and profilin.\",\n      \"method\": \"GST pulldown with recombinant Nck from BJAB cell lysates, domain mapping with WIP fragments\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct pulldown with domain mapping, single lab, two orthogonal methods (precipitation + co-IP of profilin)\",\n      \"pmids\": [\"9694849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The yeast WIP homologue End5p/verprolin interacts with the WASp homologue Las17p (yeast ortholog of WASP); high-copy LAS17 partially suppresses end5-1 growth and endocytosis defects, establishing that the WIP–WASP functional partnership is conserved in yeast and essential for endocytosis.\",\n      \"method\": \"Yeast two-hybrid, high-copy suppressor screen, endocytosis assay in yeast\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis (suppressor screen) plus two-hybrid binding, replicated concept in multiple organisms\",\n      \"pmids\": [\"9742397\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Human WIP functionally complements yeast vrp1 (verprolin) mutations, restoring cytoskeletal organization and endocytosis; this complementation requires the WH2 actin-binding domain and the profilin-binding domain of WIP, establishing these as the functional core of WIP's role in cell polarity and actin organization.\",\n      \"method\": \"Yeast complementation assay, WIP domain mutants, immunofluorescence localization in yeast\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — reconstituted function in yeast with domain mapping, direct localization, single lab\",\n      \"pmids\": [\"10358064\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"WIP mediates recruitment of N-WASP to vaccinia virus actin tails via the WH1 domain of N-WASP (not the polyproline region). For Shigella, N-WASP recruits WIP. The N-WASP–WIP complex integrates signaling cascades (SH2/SH3-adaptor pathway and Cdc42 pathway) leading to Arp2/3-dependent actin polymerization and pathogen actin-based motility.\",\n      \"method\": \"Cell-based actin tail assays with domain mutants, co-localization, dominant-negative experiments\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple genetic/biochemical approaches, two distinct pathogen models, independently replicated concept\",\n      \"pmids\": [\"10878810\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"WIP directly interacts with N-WASP and with actin; WIP retards N-WASP/Cdc42-activated actin polymerization by the Arp2/3 complex and stabilizes actin filaments. Microinjection of WIP induces filopodia in an N-WASP-dependent manner; anti-WIP antibody blocks filopodium induction by bradykinin, Cdc42(V12), and N-WASP, establishing WIP and N-WASP as a functional unit in filopodium formation.\",\n      \"method\": \"In vitro actin polymerization assay (pyrene-actin), microinjection with function-blocking antibodies, immunofluorescence\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro reconstitution assay plus microinjection functional experiments, multiple orthogonal methods\",\n      \"pmids\": [\"11331876\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"Rat WIP co-immunoprecipitates with N-WASP in vivo and co-localizes with actin stress fibers. Co-expression of WIP and N-WASP redistributes N-WASP from the nucleus to perinuclear/actin-associated locations and dissolves stress fibers while promoting filopodia formation, indicating WIP controls N-WASP subcellular localization.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, co-expression in fibroblasts and tumor cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP plus localization imaging, single lab, two orthogonal methods\",\n      \"pmids\": [\"11687573\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"NMR structure of the N-WASP EVH1 (WH1) domain in complex with a 25-residue WIP motif revealed a novel recognition mechanism: the WIP peptide wraps around the EVH1 domain contacting an extended surface, mechanistically explaining how WAS missense mutations in this domain disrupt WIP binding.\",\n      \"method\": \"NMR structure determination with WIP peptide\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — high-resolution NMR structure with direct functional correlation to disease mutations\",\n      \"pmids\": [\"12437929\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"WIP-deficient mice have T cells that fail to proliferate, secrete IL-2, increase F-actin content, polarize, or form immune synapses after TCR ligation, establishing WIP as essential for T cell activation and immunological synapse formation. WIP-deficient B cells show enhanced proliferation, indicating differential roles in T vs B cells.\",\n      \"method\": \"WIP knockout mouse model, T cell proliferation/activation assays, F-actin staining, conjugate formation assay\",\n      \"journal\": \"Immunity\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic KO with multiple defined cellular phenotype readouts, replicated across several studies\",\n      \"pmids\": [\"11869681\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Phosphatidylinositol 4,5-bisphosphate (PIP2)-induced vesicle motility requires N-WASP and involves WIP recruitment alongside Nck and Grb2; reconstitution in N-WASP-defective cells with mutants showed that both the WH1 domain (which recruits WIP) and the polyproline domain contribute significantly to N-WASP recruitment/activation at vesicle surfaces.\",\n      \"method\": \"Reconstitution of vesicle motility in N-WASP-deficient cells with domain mutants, fluorescence microscopy\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — reconstitution assay with multiple domain mutants, single lab\",\n      \"pmids\": [\"12147689\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"Cortactin SH3 domain interacts with WIP in an SH3-dependent manner (GST pulldown). WIP increases the efficiency of cortactin-mediated Arp2/3 complex activation in a concentration-dependent manner, and co-expression of cortactin and WIP stimulates membrane protrusions.\",\n      \"method\": \"Yeast two-hybrid, GST-cortactin pulldown, in vitro Arp2/3 actin polymerization assay, overexpression/localization\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro Arp2/3 assay plus binding studies and cellular readout, single lab multiple orthogonal methods\",\n      \"pmids\": [\"12620186\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"WIP participates in PDGF-induced ruffle formation: overexpression enhances ruffling, microinjection of anti-WIP antibody or WIP deficiency decreases ruffling, and a WIP mutant lacking the actin-binding site blocks PDGF-induced membrane ruffling in murine fibroblasts, establishing WIP's actin-binding domain as required for ruffle formation downstream of PDGF/Rac1.\",\n      \"method\": \"Microinjection of anti-WIP antibody, WIP overexpression, domain mutant (lacking actin-binding site), immunofluorescence, video microscopy in 3T3 and WIP-/- fibroblasts\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — antibody microinjection, KO cells, and domain mutant with functional readout, multiple orthogonal approaches\",\n      \"pmids\": [\"12724353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Toca-1 (a PCH/F-BAR protein) binds both N-WASP and Cdc42 and activates the N-WASP–WIP complex (the predominant form of N-WASP in cells) to promote actin nucleation; two distinct Cdc42 effectors (N-WASP–WIP and Toca-1) cooperate and are both required for Cdc42-induced actin assembly.\",\n      \"method\": \"Biochemical purification of Toca-1, in vitro actin assembly assays, co-IP, depletion experiments\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — biochemical purification plus in vitro reconstitution, multiple orthogonal methods, widely replicated\",\n      \"pmids\": [\"15260990\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"WIP-deficient mast cells show impaired degranulation, IL-6 secretion, calcium mobilization, and reduced phosphorylation of Syk, PLCγ2, and JNK after FcεRI ligation. WIP co-immunoprecipitates with Syk after FcεRI ligation and inhibits Syk degradation, establishing WIP as a regulator of FcεRI signaling via maintenance of Syk levels.\",\n      \"method\": \"WIP-/- bone marrow-derived mast cells, degranulation assay, co-immunoprecipitation with Syk, immunoblotting for Syk levels\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO model with specific molecular mechanism (Syk association and stability), co-IP validation\",\n      \"pmids\": [\"14757742\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"WIP is essential for podosome formation in dendritic cells (DCs): WIP-/- DCs cannot form actin cores containing WASP and cortactin. WIP regulates podosome structure by controlling calpain-mediated cleavage of WASP and by facilitating WASP localization to actin polymerization sites at podosomes.\",\n      \"method\": \"WIP-/- DCs, immunofluorescence, live imaging, calpain inhibitor experiments\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with specific molecular mechanism (calpain regulation of WASP), multiple readouts, two orthogonal approaches\",\n      \"pmids\": [\"17141616\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"WIP, WASp, actin, and myosin IIA form a multiprotein complex (~1.3 MDa) in activated NK cells. Inhibitory KIR signaling decreases actin and myosin IIA recruitment to the constitutive WIP–WASp complex. PKCθ-mediated phosphorylation of WIP correlates with increased complex formation. WIP knockdown inhibits NK cell cytotoxicity.\",\n      \"method\": \"Co-immunoprecipitation/gel filtration, RNAi, kinase inhibitor studies in YTS NK cell line\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multi-protein complex identified by co-IP/fractionation plus functional RNAi readout, single lab multiple methods\",\n      \"pmids\": [\"16606694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"WIP acts as a chaperone for WASP: WASP protein (but not mRNA) levels are severely reduced in T cells from WIP-/- mice and restored by WIP re-introduction. The WIP WASP-binding domain protects WASP from calpain-mediated degradation in vitro. Proteasome inhibitors increase WASP levels in WIP-deficient cells, indicating WASP is degraded by both calpain and the proteasome when unbound from WIP.\",\n      \"method\": \"WIP-/- mouse T cells, immunoblotting for WASP, in vitro calpain degradation assay, proteasome inhibitor treatment (MG132, bortezomib), calpain inhibitor (calpeptin)\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro degradation assay plus genetic KO plus pharmacological rescue, multiple orthogonal methods, replicated by multiple labs\",\n      \"pmids\": [\"17213309\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Drosophila D-WIP (WIP ortholog) is expressed specifically in myoblasts and bridges the WASp-Arp2/3 actin nucleation system to the myoblast adhesion molecules Dumbfounded and Sticks and Stones, recruiting the actin polymerization machinery to fusion sites. Loss of D-WIP or Wsp blocks myoblast fusion at the stage of fusion pore enlargement.\",\n      \"method\": \"Drosophila genetic analysis, immunoprecipitation, immunofluorescence in embryos\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis in Drosophila ortholog plus biochemical interaction, defined cellular phenotype\",\n      \"pmids\": [\"17419994\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"WASP expression requires WIP: WASP gene transfer yields high WASP expression only when WIP is co-expressed in K562 cells; WIP knockdown in T cells reduces WASP levels. The minimal WIP region that rescues WASP expression is the WASP-binding domain, but this minimal domain is insufficient to rescue WASP-dependent NFAT-mediated IL-2 transcription.\",\n      \"method\": \"Co-expression in K562 cells, siRNA knockdown in T cells, domain-mapping WIP mutants, reporter assay\",\n      \"journal\": \"International immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi plus re-expression plus domain mapping, single lab, multiple orthogonal approaches\",\n      \"pmids\": [\"17205972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"WIP–WASP complex mediates TCR-induced NFAT activation without dissociation: PKCθ-mediated phosphorylation of WIP Ser488 does not cause WIP–WASP dissociation; WIP–WASP complexes persist after TCR stimulation; a WIP–WASP fusion protein efficiently mediates NFAT activation. The WIP N-terminus (polyproline and WH2 domain) is inhibitory for TCR-mediated NFAT activation.\",\n      \"method\": \"Co-IP after TCR stimulation, WIP-WASP fusion protein, domain truncation, NFAT reporter assay in Jurkat cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple WIP mutants and fusion constructs, co-IP, reporter assay, single lab\",\n      \"pmids\": [\"17711847\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"WIP is indispensable for NK cell cytotoxicity: WIP knockdown completely inhibits cytolysis; WIP overexpression enhances cytolytic ability. WIP co-localizes with lytic granules and segregates to the lysosomal fraction. WIP knockdown inhibits polarization of lytic granules to the immune synapse (but not conjugate formation), and granule-WIP interaction is independent of WASp.\",\n      \"method\": \"RNAi knockdown and overexpression in YTS NK cells, cytotoxicity assays, subcellular fractionation, immunofluorescence co-localization\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi plus overexpression plus fractionation, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"18258743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"EFC/F-BAR proteins (FBP17 and Toca-1) activate N-WASP–WIP complex-mediated actin polymerization in a membrane curvature-dependent manner, requiring phosphatidylserine-containing membranes and Toca-1/FBP17 but not Cdc42 or PIP2. Toca-1/FBP17 recruit N-WASP–WIP to the membrane via conserved acidic residues near their SH3 domains.\",\n      \"method\": \"In vitro actin polymerization assays with liposomes of defined curvature, domain mutants\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution with defined membrane curvature plus domain mapping, single lab\",\n      \"pmids\": [\"18923421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"WIP is essential for IL-2 signaling in T cells: WIP/WASP double-KO (DKO) T cells (unlike WASP-KO alone) fail to respond to IL-2, as evidenced by failure to up-regulate CD25, phosphorylate STAT5, or induce STAT5-dependent genes after antigen stimulation. DKO T cells have a disrupted subcortical actin cytoskeleton and impaired TCR-triggered actin polymerization.\",\n      \"method\": \"WIP/WASP double-KO mouse model, IL-2 signaling assays (STAT5 phosphorylation, CD25 up-regulation), F-actin content measurements\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean double-KO genetic model with multiple signaling readouts compared to single KO controls\",\n      \"pmids\": [\"19359486\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The cortactin-binding domain of WIP (residues 110–170) is essential for podosome formation and MMP-mediated extracellular matrix degradation by dendritic cells; WIP-/- DCs can synthesize MMPs but fail to degrade matrix. Lentiviral rescue with WIPΔ110–170 restores disorganized podosomes but not matrix degradation.\",\n      \"method\": \"WIP-/- DCs, lentiviral rescue with domain deletion mutant, gelatin degradation assay, immunofluorescence for MMP localization\",\n      \"journal\": \"European journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with domain-specific rescue, functional matrix degradation readout, single lab\",\n      \"pmids\": [\"20952093\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Blown fuse (Blow), an FCM-specific Drosophila protein, modulates the stability of the WASP–WIP complex by competing with WASP for WIP binding; this competition drives rapid exchange of WASP, WIP, and G-actin within the podosome-like structure, which is required for fusion pore formation in myoblast fusion.\",\n      \"method\": \"Drosophila genetics, biochemical competition assays, co-IP, FRAP\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic and biochemical competition experiments, defined cellular process, Drosophila ortholog\",\n      \"pmids\": [\"21571220\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"WIP deficiency caused by a homozygous WIPF1 stop codon mutation (c.1301C>G) results in undetectable WASP protein (despite normal WAS mRNA), establishing that WIP stabilizes WASP in human T cells. Introduction of WIP into patient T cells restored WASP expression.\",\n      \"method\": \"Patient cells with WIPF1 mutation, immunoblotting for WASP, lentiviral WIP re-introduction\",\n      \"journal\": \"The Journal of experimental medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — natural human genetic experiment with molecular rescue, replicated mechanistic concept established in mice\",\n      \"pmids\": [\"22231303\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"WIP (or its homolog WIRE) is an essential link between Nck and N-WASP for Arp2/3-dependent actin assembly: N-WASP recruitment to vaccinia virus depends on WIP (not on direct Nck–N-WASP interaction). WIP contains two Nck-binding sites and is recruited to virus by the second SH3 domain of Nck while bound to N-WASP. The first and third SH3 domains of Nck are required to stimulate actin assembly but not to recruit the WIP:N-WASP complex.\",\n      \"method\": \"MEFs lacking Nck, WIP, or N-WASP; vaccinia actin-tail assay; domain mutants of Nck and WIP; co-IP\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — triple-KO MEF analysis plus domain mapping plus functional actin assembly assay, multiple orthogonal approaches\",\n      \"pmids\": [\"23707428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WIP and WASp form two distinct molecular interfaces in cells: (i) WH1 domain of WASp with C-terminal WIP, dependent on PKCθ-mediated WIP phosphorylation (in response to TCR activation); (ii) VCA domain of WASp with N-terminal WIP, dependent on actin (inhibited by latrunculin A). WASp activation involves dissociation of interface (i) while interface (ii) remains, exposing the WASp ubiquitylation site and promoting degradation.\",\n      \"method\": \"Triple-color FRET (3FRET) in live T cells, PKCθ inhibitors, latrunculin A, phosphomimetic WIP mutants\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — novel 3FRET technology with pharmacological validations and mutant controls, single lab multiple orthogonal methods\",\n      \"pmids\": [\"24962707\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Tyrosine phosphorylation of WIP (mediated by Bruton's tyrosine kinase, Btk) releases bound WASP from the WIP–WASP complex; in the absence of WIP–WASP binding, WASP is rapidly degraded. WIP phosphomimics abolish WIP–WASP interaction and disrupt podosomes; WIP lacking tyrosine phosphorylation extends podosome lifetimes. Btk was identified as a kinase regulating WIP tyrosine phosphorylation.\",\n      \"method\": \"WIP knockdown, phosphomimic/phosphonull WIP mutants, kinase screen with inhibitors, podosome assay, matrix degradation assay in macrophages\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphomimetic domain mutants, kinase identification, functional podosome/degradation readouts, single lab multiple methods\",\n      \"pmids\": [\"25413351\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WIP binding to actin (via its actin-binding domain, ABD), independently of its binding to WASp, is critical for the integrity of the actin cytoskeleton in T cells and for their migration; WIPΔABD mice have T cells with normal WASp levels but decreased F-actin, disorganized actin cytoskeleton, impaired chemotaxis, and defective homing to lymph nodes.\",\n      \"method\": \"Knock-in mice expressing WIP lacking the ABD (WIPΔABD), F-actin staining, chemotaxis assays, lymph node homing assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean knock-in mouse model isolating actin-binding from WASp-binding, multiple functional readouts\",\n      \"pmids\": [\"25246631\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WIP binding to F-actin (via ABD) is required for focal adhesion assembly and stress fiber formation in fibroblasts. WIP-/- fibroblasts have defective focal adhesions, increased G-actin levels, and reduced nuclear MRTF-A/SRF activity; constitutively nuclear MRTF-A or active SRF restores these defects, establishing a WIP–actin–MRTF–SRF axis in cell adhesion.\",\n      \"method\": \"WIP-/- fibroblasts, knock-in WIP mutant (fails to bind actin), MRTF-A nuclear translocation, SRF reporter, focal adhesion immunostaining\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO plus knock-in domain mutant plus downstream transcription factor rescue, multiple orthogonal methods\",\n      \"pmids\": [\"24797074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"NMR structural characterization showed WIP N-terminal ABM is intrinsically disordered but has residual helical (residues 30–42) and β-strand (residues 44–62) propensities that echo the actin-bound conformation; residues 17–25 preceding the canonical ABM also show β-strand propensity, suggesting the WIP–actin interaction epitope extends to the N-terminal polyproline region.\",\n      \"method\": \"NMR (protonless 13C'-detected spectroscopy), secondary chemical shifts, RDC measurements\",\n      \"journal\": \"The FEBS journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — structural NMR data in isolated domain, no functional mutagenesis validation, single lab\",\n      \"pmids\": [\"25495558\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Intersectin adaptor proteins ITSN1 and ITSN2 interact with WIP via SH3 domain–proline-rich motif interactions (middle part of WIP proline-rich region). ITSN1, WIP, and N-WASP form a trimeric complex in cells. Endogenous ITSN1 co-localizes with WIP at invadopodia in breast cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, GST-SH3 pulldown, immunofluorescence co-localization in MDA-MB-231 cells\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP plus domain mapping plus co-localization, single lab\",\n      \"pmids\": [\"25797047\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"DOCK8 is connected to WASp and actin in T cells through WIP acting as a bridge: WIP co-immunoprecipitates with DOCK8 and WASp. DOCK8 guanine nucleotide exchange factor activity is essential for WASp activation, F-actin assembly, immune synapse/actin foci formation, mechanotransduction, T cell transendothelial migration, and lymph node homing—all of which also depend on WASp, placing DOCK8 and WASp in the same actin-regulatory pathway via WIP.\",\n      \"method\": \"Co-immunoprecipitation, T cell functional assays (immune synapse, foci, migration), T cells from DOCK8-deficient and WAS patients\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP establishing DOCK8–WIP–WASp complex plus multiple functional assays in human and mouse primary cells\",\n      \"pmids\": [\"27599296\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WIP and WICH/WIRE play non-redundant roles in invadopodium formation in breast cancer cells: WIP interacts with N-WASP and cortactin and is essential for invadopodium assembly, while WICH/WIRE regulates N-WASP activation to control invadopodium maturation and degradative activity. Nck interaction with WIP modulates invadopodium maturation.\",\n      \"method\": \"RNAi knockdown of WIP and WICH/WIRE, co-immunoprecipitation, invadopodium formation/matrix degradation assays, TIRF microscopy\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RNAi with rescue, co-IP, functional invadopodium assay, multiple orthogonal methods\",\n      \"pmids\": [\"27009365\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WIP controls tumor growth by stabilizing the YAP/TAZ complex via the endocytic/endosomal system: when WIP levels are high, the β-catenin destruction complex (APC–axin–GSK3) is sequestered to multi-vesicular body compartments, inhibiting YAP/TAZ degradation. YAP/TAZ stability is dependent on Rac, PAK, and mDia, and is Hippo-independent.\",\n      \"method\": \"WIP knockdown/overexpression in cancer cells, subcellular fractionation, co-IP for destruction complex, Rac/PAK/mDia inhibitors, in vivo xenograft\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — fractionation plus co-IP plus pharmacological inhibitors plus in vivo, single lab\",\n      \"pmids\": [\"27851961\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mutant p53 oncogenic activity is driven by WIP: WIP is phosphorylated by AKT2 downstream of mtp53/p63-enhanced PI3K/AKT2-mediated integrin/receptor recycling pathways. WIP regulates YAP/TAZ stability; WIP knockdown reduces CSC markers (CD133, CD44, YAP/TAZ) and tumor growth in vivo.\",\n      \"method\": \"WIP knockdown/overexpression, mtp53 overexpression, co-IP for AKT2-WIP interaction, in vivo tumor growth assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP showing AKT2–WIP interaction plus phosphorylation plus functional knockdown with in vivo validation, single lab\",\n      \"pmids\": [\"28166194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NMR and FRET analysis of the WIP C-terminal (residues 442–492)–WASp (residues 20–158) complex revealed a pleckstrin homology-like domain with mixed α/β fold; WIP residues 454–456 are the major contributor to WASp affinity, and residues 449–451 have the largest effect on WASp ubiquitylation and degradation. WIP binding to WASp is inversely linked to WASp ubiquitylation.\",\n      \"method\": \"NMR structure of complex, FRET in vivo, biochemical ubiquitylation assays, WIP peptide mutants\",\n      \"journal\": \"ACS chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure plus FRET plus ubiquitylation assay, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"29215267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"At yeast endocytic sites, WASP and WIP accumulate to a threshold level through multivalent SH3 domain–PRM interactions involving linker proteins; Arp2/3-mediated actin assembly initiation is tightly coupled to reaching threshold levels of WASP and WIP (not to recruitment kinetics or autoinhibition release), giving actin assembly onset switch-like behavior.\",\n      \"method\": \"Quantitative live-cell fluorescence imaging of endocytic sites, yeast genetics (SH3/PRM mutants)\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — quantitative imaging with genetic perturbations in yeast ortholog system, single lab\",\n      \"pmids\": [\"28813247\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"WIP interacts with RhoA: in lung adenocarcinoma cells, WIP knockdown reduces RhoA levels and WIP co-immunoprecipitates with RhoA; WIP regulates invasion, EMT, and anchorage-independent growth via RhoA.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, invasion assays, RhoA immunoblotting\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single co-IP and knockdown, single lab, cancer cell line only\",\n      \"pmids\": [\"27939884\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"WIP/ITSN1 complex co-localizes with RAB4-positive fast-recycling endosomes and is involved in transferrin receptor recycling. ITSN1 recruits WIP to RAB4-positive vesicles. WIP enhances N-WASP–ITSN1 interaction and ITSN1/β-actin association, and the WIP/ITSN1-L complex promotes filopodia-like protrusion formation.\",\n      \"method\": \"Co-immunoprecipitation, transferrin recycling assay, co-localization with Rab4, overexpression in MCF-7 cells\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — co-IP and co-localization, single lab, no mechanistic dissection of which WIP domain mediates vesicle function\",\n      \"pmids\": [\"29958948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In C. elegans intestine, WIP-1 promotes scission of clathrin-coated pits by directly binding G-actin (independent of WSP-1/WASP); the cortactin-binding domain of WIP-1 serves as the binding interface for DBN-1 (Abp1), and DBN-1–F-actin interaction is essential for Dynamin-1 (DYN-1) recruitment at endocytic sites.\",\n      \"method\": \"RNAi knockdown, live imaging of CCP scission in C. elegans intestine, domain mapping for WIP-1–DBN-1 interaction\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — C. elegans ortholog, RNAi, live imaging, domain-specific interaction, WASP-independent mechanism established\",\n      \"pmids\": [\"31118234\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"WIP depletion increases reactive oxygen species and reduces NRF2 levels in glioblastoma cells. WIP stabilizes NRF2 by restraining KEAP1 E3 ligase activity; increased KEAP1 activity in WIP-depleted cells depends on actin cytoskeleton organization (via KEAP1–F-actin binding), not protection of KEAP1 from autophagic degradation.\",\n      \"method\": \"WIP knockdown, KEAP1 knockdown, NRF2ΔETGE overexpression, ROS measurements, co-IP of KEAP1–F-actin\",\n      \"journal\": \"Antioxidants (Basel, Switzerland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic rescue experiments plus co-IP identifying KEAP1–actin connection, single lab\",\n      \"pmids\": [\"32825452\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"WIPF1 interacts with ACTN4 to regulate podosome formation, matrix degradation, and actin polymerization in extravillous trophoblasts (EVTs); the ARG54 site of WIPF1 is implicated in this interaction. WIPF1 knockdown impairs EVT cell migration and trophoblast differentiation; WIPF1 is downregulated in RSA patient EVTs.\",\n      \"method\": \"Co-immunoprecipitation, WIPF1 knockdown in hTSC-derived EVTs, podosome/matrix degradation assays, site-directed mutagenesis (ARG54)\",\n      \"journal\": \"Genes & diseases\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with mutagenesis plus functional podosome/migration assay, single lab\",\n      \"pmids\": [\"40821124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"WIPF1 promotes gastric cancer cell proliferation, invasion, and migration in a myocardin (MYOCD)-dependent manner by activating the PI3K/AKT signaling pathway; MYOCD transactivates WIPF1 transcription and silencing WIPF1 significantly represses PI3K/AKT activation.\",\n      \"method\": \"siRNA knockdown, overexpression, in vitro and xenograft assays, PI3K/AKT pathway immunoblotting, MYOCD rescue experiments\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — knockdown/overexpression with pathway readout, single lab, mechanism not dissected at molecular level\",\n      \"pmids\": [\"38026208\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"C. elegans WIP-1 physically interacts with WSP-1 (WASP/N-WASP homolog) by yeast two-hybrid. RNAi knockdown of wip-1 decreases WSP-1 protein levels (not mRNA), and wsp-1 RNAi decreases WIP-1 protein levels (not mRNA), establishing mutual protein stabilization. WIP-1 RNAi causes embryonic lethality with hypodermal cell migration defects (ventral enclosure) similar to wsp-1 RNAi.\",\n      \"method\": \"Yeast two-hybrid, RNAi, Western blot, immunostaining in C. elegans embryos\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — ortholog functional studies with yeast two-hybrid, RNAi, protein level quantitation, and defined morphogenetic phenotype\",\n      \"pmids\": [\"16378591\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WIP-deficient hippocampal neurons show enlarged somas, overgrowth of neuritic and dendritic branches at early developmental stages, increased dendritic arborization, and increased amplitude and frequency of miniature excitatory postsynaptic currents, identifying WIP as a negative regulator of neuronal maturation and synaptic activity.\",\n      \"method\": \"WIP-/- mouse hippocampal neurons, morphometric analysis, electrophysiology (mEPSC recording)\",\n      \"journal\": \"Cerebral cortex (New York, N.Y. : 1991)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO with quantitative morphology and electrophysiology, two orthogonal readouts, single lab\",\n      \"pmids\": [\"21810783\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WIP absence in dendritic spines increases spine size and F-actin levels through a RhoA/ROCK/profilinIIa-dependent (N-WASP/Arp2/3-independent) mechanism. WIP deficiency causes transcriptional upregulation of neutral sphingomyelinase (NSM) via active RhoA, reducing membrane sphingomyelin, which in turn enhances RhoA membrane association and raft partitioning. NSM inhibition or sphingomyelin addition reverses RhoA, F-actin, and functional anomalies in WIP-/- synapses.\",\n      \"method\": \"WIP-/- mouse neurons, pharmacological inhibition of NSM, sphingomyelin supplementation, RhoA localization assay, F-actin staining, Arp2/3 co-IP\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO plus pharmacological rescue plus lipid supplementation, single lab, novel mechanism\",\n      \"pmids\": [\"24698977\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WIPF1 (WIP) is an intrinsically disordered, multifunctional scaffold protein that binds WASP/N-WASP (through its C-terminal WASp-binding domain), actin (through its N-terminal WH2/ABD domain), profilin, Nck, cortactin, DOCK8, and other SH3-containing adaptors, thereby acting as a core hub that (i) stabilizes WASP/N-WASP from calpain- and proteasome-mediated degradation, (ii) recruits and positions the N-WASP–Arp2/3 actin nucleation machinery to sites of actin polymerization (immune synapses, podosomes, invadopodia, filopodia, endocytic pits, pathogen tails), (iii) directly stabilizes F-actin filaments, (iv) integrates upstream signals (Cdc42 via Toca-1/FBP17, Nck, PKCθ phosphorylation, Btk-mediated tyrosine phosphorylation, AKT2) to regulate when and where WASP is activated or degraded, and (v) exerts WASP-independent functions including lytic granule polarization in NK cells, MRTF-SRF-driven focal adhesion assembly, YAP/TAZ stabilization through endosomal sequestration of the destruction complex, and regulation of NRF2 oxidative-stress response through actin-dependent KEAP1 restraint.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"WIPF1 (WIP) is an intrinsically disordered scaffold protein that couples actin nucleation machinery to upstream signals and controls the stability and localization of WASP/N-WASP at sites of actin polymerization [#0, #5]. It was first identified through its direct interaction with WASP, binding at a site distinct from the Cdc42-binding region while carrying its own N-terminal actin- and profilin-binding motifs required to drive actin polymerization [#0]; the structural basis of the C-terminal interaction was resolved as a peptide that wraps around the N-WASP EVH1/WH1 domain, explaining how WAS disease mutations disrupt WIP binding [#7, #37]. A central function of WIP is to act as a chaperone that protects WASP from calpain- and proteasome-mediated degradation: WASP protein but not mRNA is lost in WIP-deficient cells and restored by re-expression, a mechanism conserved from C. elegans through mouse to a human WIPF1 stop-codon patient in whom WASP was undetectable [#16, #25, #45]. WIP positions and activates the N-WASP–Arp2/3 system at diverse actin structures, integrating signals from Nck, cortactin, the F-BAR proteins Toca-1/FBP17 in a membrane-curvature-dependent manner, and DOCK8 [#1, #10, #12, #21, #26, #33]. Through these activities WIP drives filopodia, membrane ruffling, pathogen actin tails, podosomes and invadopodia with associated matrix degradation, and clathrin-mediated endocytosis [#4, #5, #11, #14, #23, #34, #38]. In the immune system WIP is essential for T-cell activation, immune-synapse formation, and IL-2 signaling, for FcεRI-driven mast cell signaling via stabilization of Syk, and for NK-cell cytotoxicity through WASP-independent polarization of lytic granules [#8, #13, #20, #22]. WIP also exerts WASP-independent functions through direct actin binding, supporting cytoskeletal integrity and T-cell migration, focal adhesion assembly via an actin–MRTF–SRF axis, and regulation of neuronal maturation and synaptic activity [#29, #30, #46]. Loss-of-function WIPF1 mutation in humans causes a WAS-like primary immunodeficiency with absent WASP [#25].\",\n  \"teleology\": [\n    {\n      \"year\": 1997,\n      \"claim\": \"Established WIP as a physical and functional partner of WASP, defining its core domain architecture (actin/profilin-binding N-terminus, distinct WASP-binding site) and its capacity to drive actin polymerization.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP from lymphocytes, and truncation-mutant expression in B cells\",\n      \"pmids\": [\"9405671\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not establish whether WIP regulates WASP stability or only localization\", \"Mechanism of actin polymerization induction not resolved biochemically\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Showed WIP links the adaptor Nck and profilin to the cytoskeleton through domain-specific interactions, placing WIP within receptor-proximal signaling adaptor networks.\",\n      \"evidence\": \"GST-Nck pulldown and WIP fragment domain mapping in B-cell lysates\",\n      \"pmids\": [\"9694849\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, no functional readout for the Nck–WIP interaction\", \"Did not test reciprocity in vivo\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Demonstrated functional conservation of the WIP–WASP partnership in endocytosis and cell polarity, mapping the requirement to the WH2 actin- and profilin-binding domains.\",\n      \"evidence\": \"Yeast genetic suppression/complementation with verprolin orthologs and WIP domain mutants\",\n      \"pmids\": [\"9742397\", \"10358064\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Yeast system may not capture mammalian-specific regulatory inputs\", \"Did not address WASP stabilization role\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Defined WIP as the recruiter of N-WASP to actin-polymerization sites via the N-WASP WH1 domain, integrating Cdc42 and SH2/SH3-adaptor pathways during pathogen actin-based motility.\",\n      \"evidence\": \"Cell-based vaccinia and Shigella actin-tail assays with domain mutants and dominant negatives\",\n      \"pmids\": [\"10878810\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve how upstream signals choose between recruitment and activation\", \"Quantitative contribution of WIP vs other recruiters unclear\"]\n    },\n    {\n      \"year\": 2001,\n      \"claim\": \"Established the biochemical activity of WIP: direct N-WASP and actin binding, F-actin stabilization, and a required role with N-WASP in filopodium formation.\",\n      \"evidence\": \"In vitro pyrene-actin assays, microinjection with function-blocking antibodies, and co-IP/imaging\",\n      \"pmids\": [\"11331876\", \"11687573\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Apparent inhibition of Arp2/3 nucleation in vitro vs activation in cells not reconciled\", \"Regulation of the WIP–N-WASP unit by phosphorylation not yet addressed\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Genetic loss-of-function established WIP as essential for T-cell activation and immune synapse formation, with cell-type-specific (T vs B) consequences, and showed WIP supports N-WASP recruitment at vesicle surfaces.\",\n      \"evidence\": \"WIP-knockout mouse T/B cell assays and N-WASP-deficient cell reconstitution with domain mutants\",\n      \"pmids\": [\"11869681\", \"12147689\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not distinguish actin-scaffolding from WASP-stabilizing contributions to the T-cell phenotype\", \"Molecular basis of the divergent B-cell phenotype unresolved\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Resolved the structural recognition mechanism between WIP and the N-WASP EVH1/WH1 domain, providing a molecular explanation for WAS-associated missense mutations that disrupt WIP binding.\",\n      \"evidence\": \"NMR structure of the N-WASP WH1 domain bound to a WIP peptide\",\n      \"pmids\": [\"12437929\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of full-length disordered WIP not determined\", \"Did not address the actin-dependent second interface\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Extended WIP's effector network to cortactin and to Rac1/PDGF-driven membrane dynamics, with the actin-binding domain required for ruffle formation.\",\n      \"evidence\": \"Cortactin SH3 pulldowns and in vitro Arp2/3 assays; anti-WIP microinjection and domain mutants in fibroblasts\",\n      \"pmids\": [\"12620186\", \"12724353\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How cortactin and N-WASP inputs are coordinated on WIP not resolved\", \"In vivo relevance of cortactin synergy to specific actin structures untested\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identified F-BAR protein Toca-1 as an activator of the predominant cellular N-WASP–WIP complex, establishing cooperative Cdc42 effector logic, and broadened WIP's signaling roles to mast cell FcεRI signaling through Syk stabilization.\",\n      \"evidence\": \"Biochemical purification and in vitro actin assembly; WIP-/- mast cells with Syk co-IP and stability assays\",\n      \"pmids\": [\"15260990\", \"14757742\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Syk stabilization is mechanistically analogous to WASP chaperoning untested\", \"Toca-1 vs FBP17 specificity not yet defined\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Established WIP as essential for podosome formation and revealed its role in regulating calpain-mediated WASP cleavage, and identified a WASP-containing actin/myosin megacomplex in NK cells modulated by PKCθ phosphorylation.\",\n      \"evidence\": \"WIP-/- dendritic cells with calpain inhibitors; co-IP/gel filtration and RNAi in NK cell lines\",\n      \"pmids\": [\"17141616\", \"16606694\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How phosphorylation controls complex composition not mechanistically resolved\", \"Link between podosome defect and WASP degradation only correlative\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defined WIP's chaperone function in molecular terms—the WASP-binding domain protects WASP from calpain and proteasomal degradation—while showing this minimal protection is insufficient for full WASP-dependent signaling, and extended conserved WIP–WASP partnership to Drosophila myoblast fusion.\",\n      \"evidence\": \"WIP-/- T cells with in vitro calpain assays and proteasome/calpain inhibitors; K562 co-expression and domain mapping; Drosophila genetics\",\n      \"pmids\": [\"17213309\", \"17205972\", \"17419994\", \"17711847\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the proteasomal E3 ligase for free WASP not defined\", \"How the WIP N-terminus inhibits NFAT activation mechanistically unclear\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Established a WASP-independent role for WIP in NK-cell cytotoxicity through lytic granule polarization, demonstrating WIP functions beyond the WIP–WASP axis.\",\n      \"evidence\": \"RNAi/overexpression in NK cell lines with cytotoxicity assays, subcellular fractionation, and co-localization with lytic granules\",\n      \"pmids\": [\"18258743\", \"18923421\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular tether linking WIP to lytic granules unidentified\", \"Membrane-curvature activation studied in vitro only\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Used a WIP/WASP double knockout to show WIP is required for IL-2/STAT5 signaling and subcortical actin integrity beyond WASP loss alone, separating WIP-specific from WASP-shared functions.\",\n      \"evidence\": \"WIP/WASP double-KO mouse T cells with STAT5 phosphorylation, CD25, and F-actin readouts versus single-KO controls\",\n      \"pmids\": [\"19359486\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct molecular link between WIP and the IL-2 receptor signaling apparatus not defined\", \"Whether actin defect causes the signaling defect untested\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Mapped the cortactin-binding region of WIP as essential for matrix-degrading function of podosomes, dissociating podosome assembly from degradative activity.\",\n      \"evidence\": \"WIP-/- dendritic cells rescued with a domain-deletion mutant and gelatin degradation assays\",\n      \"pmids\": [\"20952093\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the cortactin-binding region directs MMP delivery not resolved\", \"Direct cortactin requirement not genetically isolated\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Revealed dynamic competitive regulation of the WASP–WIP complex (by Blow in Drosophila) driving rapid component exchange required for fusion-pore formation, and identified WIP as a negative regulator of neuronal maturation and synaptic activity.\",\n      \"evidence\": \"Drosophila genetics with competition and FRAP assays; WIP-/- mouse hippocampal neuron morphometry and electrophysiology\",\n      \"pmids\": [\"21571220\", \"21810783\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether mammalian factors analogous to Blow regulate WASP–WIP exchange unknown\", \"Molecular pathway underlying the neuronal phenotype not yet defined here\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Confirmed in humans that WIPF1 loss-of-function abolishes WASP protein and causes a WAS-like immunodeficiency, validating the chaperone model as clinically relevant.\",\n      \"evidence\": \"Patient cells with a homozygous WIPF1 stop-codon mutation, WASP immunoblotting, and lentiviral WIP rescue\",\n      \"pmids\": [\"22231303\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Genotype-phenotype range of WIPF1 deficiency not delineated\", \"Did not address WASP-independent contributions to patient phenotype\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Established WIP (or WIRE) as the obligatory link between Nck and N-WASP for Arp2/3-dependent actin assembly, separating the recruitment step from the actin-stimulation step within Nck.\",\n      \"evidence\": \"Nck-, WIP-, and N-WASP-deficient MEFs in vaccinia actin-tail assays with Nck/WIP domain mutants\",\n      \"pmids\": [\"23707428\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the first/third Nck SH3 domains stimulate assembly mechanistically unresolved\", \"Generality beyond pathogen tails to physiological structures not tested\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined two distinct WIP–WASP interfaces (a phosphorylation-dependent WH1 interface and an actin-dependent VCA interface) and showed WASP activation couples interface remodeling to exposure of its ubiquitylation site, mechanistically linking activation to degradation; also isolated WIP's WASP-independent actin-binding functions in T-cell migration and focal-adhesion/MRTF–SRF signaling, and resolved the disordered actin-binding motif by NMR.\",\n      \"evidence\": \"Triple-color FRET in live T cells with PKCθ inhibitors/phosphomimetics; Btk kinase identification with phosphomutants; WIPΔABD knock-in mice; WIP-/- fibroblasts with MRTF/SRF rescue; NMR of the ABM\",\n      \"pmids\": [\"24962707\", \"25413351\", \"25246631\", \"24797074\", \"25495558\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How phospho- and actin-dependent interfaces are temporally coordinated in vivo not fully resolved\", \"Identity of the WASP ubiquitin ligase still unaddressed\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Extended the WIP scaffold to intersectin adaptors, forming an ITSN1–WIP–N-WASP trimer that localizes to invadopodia.\",\n      \"evidence\": \"Co-IP, GST-SH3 pulldown, and co-localization in breast cancer cells\",\n      \"pmids\": [\"25797047\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional requirement of ITSN1 for invadopodia not tested here\", \"Single lab, no reciprocal in vivo validation\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Bridged DOCK8 to WASP-dependent actin regulation via WIP in T cells, distinguished non-redundant WIP versus WIRE roles in invadopodium assembly versus maturation, and uncovered a WASP-independent oncogenic role for WIP in stabilizing YAP/TAZ via endosomal sequestration of the β-catenin destruction complex.\",\n      \"evidence\": \"Co-IP and functional T-cell assays in DOCK8/WAS patient cells; RNAi with invadopodium assays; WIP knockdown/overexpression with fractionation, inhibitors, and xenografts\",\n      \"pmids\": [\"27599296\", \"27009365\", \"27851961\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of endosomal sequestration of the destruction complex not resolved at molecular detail\", \"How DOCK8 GEF activity is coupled to the WIP scaffold unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Provided high-resolution structure of the WIP C-terminus–WASp complex linking specific residues to WASp affinity and ubiquitylation, and placed WIP downstream of AKT2 in mutant-p53-driven tumorigenesis controlling YAP/TAZ and cancer stem-cell markers.\",\n      \"evidence\": \"NMR/FRET of the WIP–WASp complex with peptide mutants and ubiquitylation assays; co-IP, phosphorylation, and in vivo tumor assays\",\n      \"pmids\": [\"29215267\", \"28166194\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural data did not capture the full intact complex in cells\", \"AKT2-WIP phosphosite functional consequences from single lab\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Implicated WIP in RAB4-positive recycling endosome trafficking with ITSN1 and in RhoA-dependent cancer cell invasion, broadening its membrane-trafficking and tumor-promoting roles.\",\n      \"evidence\": \"Co-IP, transferrin recycling and co-localization assays in MCF-7; co-IP and invasion assays in lung adenocarcinoma\",\n      \"pmids\": [\"29958948\", \"27939884\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Co-IP/co-localization only without domain dissection of vesicle function\", \"RhoA interaction shown by single co-IP without reciprocal validation\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Demonstrated a WASP-independent endocytic function of WIP-1 in scission of clathrin-coated pits via direct G-actin binding and cortactin-domain-mediated recruitment of DBN-1/Abp1 and dynamin.\",\n      \"evidence\": \"RNAi and live imaging in C. elegans intestine with domain mapping of the WIP-1–DBN-1 interaction\",\n      \"pmids\": [\"31118234\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Conservation of this WASP-independent scission role in mammals untested\", \"How G-actin binding triggers scission mechanistically unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Connected WIP to the NRF2 oxidative-stress response, showing WIP restrains KEAP1 E3 activity through actin-cytoskeleton-dependent KEAP1–F-actin binding rather than autophagic protection.\",\n      \"evidence\": \"WIP/KEAP1 knockdown, NRF2 mutant overexpression, ROS measurement, and KEAP1–F-actin co-IP in glioblastoma cells\",\n      \"pmids\": [\"32825452\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab, no in vivo confirmation\", \"Direct mechanism by which actin organization controls KEAP1 activity unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended WIPF1 actin functions to extravillous trophoblast podosomes via ACTN4 interaction and to MYOCD-driven PI3K/AKT signaling in gastric cancer, implicating WIPF1 in trophoblast differentiation/disease and tumor progression.\",\n      \"evidence\": \"Co-IP with ARG54 site mutagenesis and migration/podosome assays in trophoblasts; knockdown/overexpression with pathway readouts and MYOCD rescue in gastric cancer\",\n      \"pmids\": [\"40821124\", \"38026208\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ACTN4 interaction from single lab without reciprocal in vivo validation\", \"Gastric cancer mechanism not dissected at molecular level\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the multiple WIP interfaces, phosphorylation inputs (PKCθ, Btk, AKT2), and partner choices are integrated in space and time to switch between WASP stabilization, WASP activation/degradation, and WASP-independent functions remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified model coupling signal input to interface remodeling in vivo\", \"E3 ligase mediating WASP/WIP-regulated ubiquitylation unidentified\", \"Structural basis of full-length WIP scaffolding not determined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 5, 29, 30, 41]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [1, 26, 33, 32]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [16, 5, 12, 37]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [16, 25, 45]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [5, 6, 29, 30]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 6]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [20, 40, 41]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [11, 14, 21]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [8, 13, 20, 22, 33]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [2, 9, 38, 41]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [12, 26, 27, 30]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [25, 35, 36]}\n    ],\n    \"complexes\": [\n      \"WIP–WASP/N-WASP complex\",\n      \"N-WASP–WIP–Arp2/3 nucleation complex\",\n      \"WIP–WASp–actin–myosin IIA megacomplex (NK cells)\",\n      \"ITSN1–WIP–N-WASP trimeric complex\"\n    ],\n    \"partners\": [\n      \"WASP\",\n      \"N-WASP\",\n      \"Nck\",\n      \"cortactin\",\n      \"Toca-1\",\n      \"DOCK8\",\n      \"ITSN1\",\n      \"profilin\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}