{"gene":"WTIP","run_date":"2026-04-28T23:00:23","timeline":{"discoveries":[{"year":2010,"finding":"WTIP (along with LIMD1 and Ajuba) localizes to processing bodies (P-bodies) and binds Ago1/2, RCK, Dcp2, and eIF4E in vivo; these LIM proteins are required for miRNA-mediated (but not siRNA-mediated) gene silencing by linking the translationally inhibited eIF4E-m7GTP-5'-cap to Ago1/2 within the miRISC complex, forming an inhibitory closed-loop complex.","method":"Co-immunoprecipitation, m7GTP cap-binding assay, P-body imaging, knockdown with reporter silencing assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, functional knockdown with specific mechanistic readout, replicated across three family members with orthogonal assays","pmids":["20616046"],"is_preprint":false},{"year":2010,"finding":"Upon LPS-induced podocyte injury, WTIP translocates from cell contacts to the nucleus via a mechanism requiring JNK activity, assembly of a JNK-interacting protein 3/dynein scaffold complex, intact microtubule networks, and dynein motor activity; loss of WTIP from cell contacts alters podocyte morphology and actin assembly.","method":"Live imaging, cellular fractionation, pharmacological inhibition (JNK inhibitor, microtubule disruption, dynein inhibition), shRNA knockdown with actin spreading assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (imaging, fractionation, pharmacological) in a single study with defined phenotypic readout","pmids":["20086015"],"is_preprint":false},{"year":2011,"finding":"WTIP localizes to focal adhesions in isolated podocytes and shifts to adherens junctions upon homotypic cell contact; WTIP knockdown disrupts actin stress fiber formation, focal adhesion maturation, and adherens junction assembly. WTIP directly interacts with the RhoA-specific GEF ARHGEF12, and WTIP-induced stress fiber formation is blocked by RhoA inhibitor C3 toxin and ROCK inhibitor.","method":"shRNA knockdown, gain-of-function overexpression, cell surface biotinylation, co-immunoprecipitation, pharmacological inhibition (C3 toxin, ROCK inhibitor)","journal":"American journal of physiology. Renal physiology","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, gain/loss-of-function with specific phenotypic readouts, pharmacological validation of pathway","pmids":["21900451"],"is_preprint":false},{"year":2009,"finding":"WTIP physically interacts with the intracellular domain of the receptor tyrosine kinase Ror2; full-length Ror2 recruits WTIP to the cell membrane, whereas a disease-associated Ror2 mutant fails to do so. WTIP inhibits canonical Wnt signaling in mammalian cells and Xenopus embryos.","method":"Co-immunoprecipitation, cellular localization assay, Xenopus Wnt reporter assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 — single Co-IP with functional Wnt inhibition assay, single lab","pmids":["19785987"],"is_preprint":false},{"year":2012,"finding":"In zebrafish, Wtip knockdown causes mitotic spindle orientation defects in the pronephros, cloaca malformation with reduced ciliated cells, and loss of striated rootlets from basal bodies resulting in absent cilia motility; Wtip and Vangl2 interact genetically (double morphant phenotype), placing Wtip in the planar cell polarity pathway.","method":"Morpholino knockdown in zebrafish, immunofluorescence, genetic epistasis (double morphant), high-speed video microscopy","journal":"Biology open","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis plus multiple phenotypic readouts, single lab","pmids":["23213452"],"is_preprint":false},{"year":2016,"finding":"Nephrin tyrosine phosphorylation recruits Nck adaptor proteins, which associate with WTIP and its binding partner Lats1, sequestering them to the nephrin complex and decreasing phospho-activation of Lats1; nephrin dephosphorylation (podocyte injury) leads to rapid Lats1 activation and subsequent YAP downregulation, connecting nephrin signaling to the Hippo pathway.","method":"Mutational analysis, co-immunoprecipitation, in vivo injury model (LPS mouse), immunoblot for phospho-Lats1/YAP","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 — mutational analysis, reciprocal Co-IP, in vivo model with defined phosphorylation readouts","pmids":["27033705"],"is_preprint":false},{"year":2016,"finding":"Prickle3 (Pk3) physically associates with Wtip and cooperates with Wtip to regulate basal body organization and ciliogenesis in Xenopus GRP; Pk3 depletion prevents γ-tubulin and Nedd1 from associating with the basal body, and Wtip interaction is required for this Pk3 function.","method":"Co-immunoprecipitation, morpholino knockdown, immunofluorescence (γ-tubulin/Nedd1 localization), Xenopus GRP assay","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus functional epistasis in Xenopus, single lab","pmids":["27062996"],"is_preprint":false},{"year":2018,"finding":"Wtip localizes to apical cell junctions in Xenopus neuroectoderm and adopts a tension-sensitive distribution (punctate vs. linear) dependent on myosin II activity; depletion of Wtip inhibits apical constriction in neuroepithelial cells causing neural tube defects. The C-terminal LIM domain fragment of Wtip physically associates with Shroom3 and inhibits Shroom3-induced apical constriction.","method":"Morpholino knockdown in Xenopus, fluorescent protein tagging/live imaging, co-immunoprecipitation, dominant-negative C-terminal fragment assay","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 — live imaging of tension-dependent localization, Co-IP, loss-of-function with defined morphogenetic phenotype","pmids":["29661847"],"is_preprint":false},{"year":2014,"finding":"WTIP directly interacts with ASXL2 and represses ASXL2-mediated transcriptional activation of retinoic acid-responsive genes; WTIP blocks ASXL2's stimulatory effect on retinoic acid signaling in luciferase reporter assays.","method":"Yeast two-hybrid, co-immunoprecipitation, luciferase reporter assay in HeLa cells","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 — Co-IP plus functional reporter assay, single lab","pmids":["25065743"],"is_preprint":false},{"year":2015,"finding":"WTIP interacts with ASXL1; Asxl1-null mouse embryos show defects in kidney size and glomerular podocyte morphology along with upregulation of Wt1/Wtip target genes, implicating the Asxl1-Wtip interaction in maintenance of podocyte structure and regulation of WT1 signaling.","method":"Co-immunoprecipitation, Asxl1 knockout mouse, gene expression analysis","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus KO mouse with defined phenotypic readout, single lab","pmids":["26385183"],"is_preprint":false},{"year":2013,"finding":"Dendrin interacts with WTIP (and Gadd45a) at the podocyte slit diaphragm, as shown by yeast two-hybrid and co-immunoprecipitation; dendrin relocalizes to the nucleus in adriamycin nephropathy, suggesting a nuclear function linked to Wtip.","method":"Yeast two-hybrid, co-immunoprecipitation, immunofluorescence in injury model","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 3 — yeast two-hybrid plus Co-IP, single lab","pmids":["24376653"],"is_preprint":false},{"year":2016,"finding":"WTIP interacts with BRCA2 (amino acids 2750–2864) in HeLa cells; WTIP knockdown abolishes BRCA2 centrosome localization and causes abnormal cell division.","method":"Yeast two-hybrid, co-immunoprecipitation, GST pull-down, shRNA knockdown, immunofluorescence microscopy","journal":"Archives of gynecology and obstetrics","confidence":"Medium","confidence_rationale":"Tier 2 — GST pull-down plus Co-IP plus KD phenotype, single lab","pmids":["27535760"],"is_preprint":false},{"year":2021,"finding":"WTIP interacts with FOXO3a, transcriptionally activates FOXO3a expression, and promotes its nuclear translocation; this induces PUMA expression and activates the intrinsic apoptotic pathway, suppressing AML cell growth in vitro and in vivo.","method":"Co-immunoprecipitation, luciferase reporter (FOXO3a transcriptional activation), overexpression/knockdown, in vivo xenograft","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus transcriptional reporter plus in vivo model, single lab","pmids":["34930905"],"is_preprint":false},{"year":2021,"finding":"The N-terminal domain of Wtip physically associates with SSX2IP (a centriolar satellite/centrosomal maturation factor); double depletion of Wtip and SSX2IP in Xenopus embryos uncovered functional interaction required for neural tube closure and cell junction remodeling.","method":"Targeted proximity biotinylation, co-immunoprecipitation, co-localization imaging, double morpholino knockdown in Xenopus","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — proximity biotinylation followed by Co-IP and genetic epistasis, single lab","pmids":["34710136"],"is_preprint":false},{"year":2018,"finding":"A UBA2-WTIP intrachromosomal fusion gene is present in AML; the fusion protein abrogates WTIP-mediated processing body (P-body) formation and induces phosphorylation of STAT3, STAT5, and ERK1/2, promoting cell proliferation.","method":"RT-PCR/Sanger sequencing, ectopic expression in KG-1a cells, immunofluorescence for P-bodies, immunoblot for pathway activation, in vivo xenograft","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — functional assays (P-body, signaling, proliferation) with in vivo validation, single lab","pmids":["30179602"],"is_preprint":false},{"year":2022,"finding":"WTIP directly interacts with ARHGEF12 (a RhoA-specific GEF) through LIM- and PDZ-binding domains with high affinity; this interaction is proposed to regulate slit diaphragm and foot process function via Rho effector signaling.","method":"Co-immunoprecipitation, domain mapping, heterozygous mouse model with LPS/adriamycin injury","journal":"American journal of physiology. Renal physiology","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP with domain mapping plus in vivo injury model, single lab","pmids":["35862649"],"is_preprint":false},{"year":2022,"finding":"WTIP overexpression in MCF10A cells reduces junctional localization of LIMD1 and LATS1, and is associated with increased YAP1 phosphorylation and decreased YAP1 nuclear localization, indicating WTIP competes with LIMD1 for adherens junction binding but cannot efficiently recruit Lats kinases, resulting in distinct (pro-Hippo) downstream effects.","method":"Overexpression in MCF10A cells, immunofluorescence for junctional localization, immunoblot for phospho-YAP1/YAP1 nuclear levels","journal":"microPublication biology","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment with functional Hippo pathway readout, single lab","pmids":["36439396"],"is_preprint":false},{"year":2022,"finding":"WTIP knockdown in neonatal rat ventricular myocytes causes cellular hypertrophy; a HCM-associated variant WTIP p.Y233F disrupts binding to a Wnt signaling protein and alters cardiomyocyte calcium dynamics (increased resting and peak calcium), which is rescued by verapamil.","method":"shRNA knockdown in NRVMs, iPSC-derived cardiomyocytes from patient, calcium imaging, morpholino knockdown in zebrafish","journal":"Circulation. Genomic and precision medicine","confidence":"Medium","confidence_rationale":"Tier 2 — multiple model systems (NRVMs, iPSC-CMs, zebrafish) with defined mechanistic readout, single lab","pmids":["35671065"],"is_preprint":false},{"year":2025,"finding":"WTIP interacts with PP2A phosphatase and inhibits T308 phosphorylation and kinase activity of AKT, thereby suppressing stemness and chemoresistance in gastric cancer; TET2 and transcription factor PATZ1 cooperate to transcriptionally activate WTIP expression.","method":"Co-immunoprecipitation (WTIP-PP2A), AKT kinase assay (T308 phosphorylation), overexpression/knockdown, TET2/PATZ1 reporter assay","journal":"Neoplasia (New York, N.Y.)","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP plus kinase activity assay with functional chemoresistance readout, single lab","pmids":["40279682"],"is_preprint":false},{"year":2016,"finding":"In zebrafish, Wtip protein localizes to the basal body of proepicardial organ (PE) cells; wtip knockdown disrupts PE specification (in conjunction with WT1 signaling), perturbs cardiac looping, and disrupts early left-right asymmetry.","method":"Morpholino knockdown in zebrafish, mRNA overexpression, immunofluorescence for basal body localization, marker gene expression","journal":"Molecular medicine reports","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization experiment with loss-of-function phenotypic readout, single lab","pmids":["27484451"],"is_preprint":false}],"current_model":"WTIP is a LIM-domain scaffold protein of the Ajuba family that functions as a tension-sensitive adaptor at cell junctions, regulating actin/actomyosin contractility through interaction with ARHGEF12 (a RhoA-GEF) and Shroom3; it undergoes JNK/dynein/microtubule-dependent nuclear translocation in response to injury to modulate WT1 and FOXO3a transcription; it suppresses Hippo signaling by linking nephrin/Nck to Lats1, modulates miRNA-mediated gene silencing via Ago1/2 and the eIF4E-m7GTP cap complex in P-bodies, interacts with Ror2 to inhibit canonical Wnt signaling, and inhibits AKT activity through PP2A, collectively coordinating cell junction integrity, cytoskeletal dynamics, and transcriptional responses in podocytes, neuroepithelium, and other tissues."},"narrative":{"teleology":[{"year":2009,"claim":"Establishing that WTIP is a Ror2-interacting protein that inhibits canonical Wnt signaling resolved the question of whether LIM-domain proteins at cell junctions could participate in Wnt pathway modulation.","evidence":"Co-immunoprecipitation and Xenopus Wnt reporter assay showing WTIP–Ror2 interaction and functional Wnt inhibition","pmids":["19785987"],"confidence":"Medium","gaps":["Single Co-IP without reciprocal validation","Endogenous interaction not confirmed","Downstream targets of WTIP-mediated Wnt inhibition not identified"]},{"year":2010,"claim":"Two contemporaneous studies established dual compartmentalization of WTIP: it localizes to P-bodies where it bridges Ago1/2 to the eIF4E–m7GTP cap for miRNA silencing, and it undergoes JNK/dynein/microtubule-dependent nuclear translocation from cell contacts upon injury, revealing stimulus-dependent shuttling as a core WTIP mechanism.","evidence":"Co-IP with cap-binding assays and knockdown reporter silencing for P-body function; live imaging, subcellular fractionation, and pharmacological inhibition for nuclear translocation","pmids":["20616046","20086015"],"confidence":"High","gaps":["Whether P-body function and nuclear translocation are independently regulated is unclear","Direct RNA targets of WTIP-dependent miRNA silencing not identified","Nuclear transcriptional targets upon injury-induced translocation not fully defined"]},{"year":2011,"claim":"Identification of ARHGEF12 as a direct WTIP partner established the mechanism by which WTIP promotes RhoA/ROCK-dependent actin stress fiber formation at focal adhesions and adherens junctions, answering how a LIM scaffold controls actomyosin contractility.","evidence":"Co-IP, shRNA knockdown, overexpression, and pharmacological inhibition with C3 toxin and ROCK inhibitor in podocytes","pmids":["21900451"],"confidence":"High","gaps":["Structural basis of WTIP–ARHGEF12 interaction not resolved","Whether WTIP activates or scaffolds ARHGEF12 catalytic activity is unknown"]},{"year":2012,"claim":"Zebrafish studies revealed WTIP functions in planar cell polarity, controlling mitotic spindle orientation and ciliogenesis through genetic interaction with Vangl2, extending WTIP's role beyond cell junctions to basal body/cilium biology.","evidence":"Morpholino knockdown in zebrafish, double morphant epistasis with Vangl2, high-speed video microscopy of cilia motility","pmids":["23213452"],"confidence":"Medium","gaps":["Morpholino-only approach without genetic mutant confirmation","Direct biochemical connection between WTIP and Vangl2 not established","Mechanism of WTIP at the basal body not defined"]},{"year":2013,"claim":"Discovery that dendrin interacts with WTIP at the podocyte slit diaphragm and relocalizes to the nucleus upon injury added another injury-responsive WTIP partner, suggesting coordinated nuclear signaling from the slit diaphragm complex.","evidence":"Yeast two-hybrid and co-immunoprecipitation, immunofluorescence in adriamycin nephropathy model","pmids":["24376653"],"confidence":"Medium","gaps":["Functional consequence of dendrin–WTIP interaction not tested","No reciprocal Co-IP reported","Whether dendrin and WTIP co-translocate to the nucleus is unknown"]},{"year":2014,"claim":"Demonstration that WTIP represses ASXL2-mediated retinoic acid signaling expanded WTIP's nuclear functions to include transcriptional corepression of chromatin regulatory complexes.","evidence":"Yeast two-hybrid, co-immunoprecipitation, luciferase reporter assay in HeLa cells","pmids":["25065743"],"confidence":"Medium","gaps":["Single Co-IP without domain mapping for ASXL2 interaction","Endogenous relevance in podocytes or other tissues not tested","Mechanism of repression (recruitment of corepressors?) not defined"]},{"year":2015,"claim":"Asxl1 knockout mice showing kidney and podocyte defects with upregulated Wt1/Wtip targets provided in vivo genetic evidence that ASXL1–WTIP interaction maintains podocyte structure.","evidence":"Co-immunoprecipitation, Asxl1 knockout mouse, gene expression analysis","pmids":["26385183"],"confidence":"Medium","gaps":["WTIP was not directly knocked out in this model","Whether WTIP is the mediating partner versus other ASXL1 targets is unresolved"]},{"year":2016,"claim":"Three parallel discoveries established WTIP as a node connecting nephrin signaling to Hippo pathway suppression (via Lats1 sequestration), cooperating with Prickle3 for basal body organization, and localizing to basal bodies in the proepicardial organ for cardiac left-right patterning.","evidence":"Nephrin mutational analysis with Co-IP and in vivo LPS model for Hippo; Pk3 Co-IP and Xenopus GRP morpholino epistasis for ciliogenesis; zebrafish morpholino KD with immunofluorescence for cardiac laterality","pmids":["27033705","27062996","27484451"],"confidence":"High","gaps":["Direct measurement of WTIP effect on Lats1 kinase activity in vitro is lacking","Whether Pk3–WTIP and nephrin–WTIP complexes overlap in vivo is unknown","Role of WTIP in proepicardial specification versus cilia function not separated"]},{"year":2016,"claim":"Identification of WTIP–BRCA2 interaction and requirement of WTIP for BRCA2 centrosome localization revealed an unexpected role in centrosome function and mitotic fidelity.","evidence":"Yeast two-hybrid, Co-IP, GST pull-down, shRNA knockdown with immunofluorescence in HeLa cells","pmids":["27535760"],"confidence":"Medium","gaps":["Single lab finding not independently replicated","Whether WTIP scaffolds BRCA2 at centrosomes or regulates its transport is unclear","Relationship to WTIP's basal body localization not explored"]},{"year":2018,"claim":"Live imaging in Xenopus showed WTIP adopts tension-sensitive distributions at apical junctions dependent on myosin II and physically associates with Shroom3 to regulate apical constriction during neural tube closure, establishing WTIP as a mechanosensitive regulator of morphogenesis.","evidence":"Morpholino knockdown in Xenopus, fluorescent protein tagging/live imaging, Co-IP, dominant-negative C-terminal fragment assay","pmids":["29661847"],"confidence":"High","gaps":["Whether tension sensing involves conformational unfolding of LIM domains is not tested","How WTIP coordinates Shroom3 and ARHGEF12 simultaneously is unresolved"]},{"year":2018,"claim":"Discovery of a UBA2-WTIP fusion in AML that abrogates P-body formation and activates STAT3/5 and ERK1/2 demonstrated pathological consequences of disrupting WTIP's scaffold functions, linking WTIP loss-of-function to oncogenic signaling.","evidence":"RT-PCR/Sanger sequencing, ectopic expression in KG-1a cells, immunofluorescence for P-bodies, immunoblot, xenograft","pmids":["30179602"],"confidence":"Medium","gaps":["Single patient fusion; frequency in AML cohorts not established","Whether STAT/ERK activation is direct or secondary to P-body loss is unclear"]},{"year":2021,"claim":"Two studies expanded WTIP's functional network: WTIP transcriptionally activates FOXO3a to induce PUMA-mediated apoptosis in AML, and the N-terminus of WTIP associates with centriolar satellite protein SSX2IP for neural tube closure, revealing both nuclear transcriptional and centrosomal functions.","evidence":"Co-IP plus luciferase reporter plus in vivo xenograft for FOXO3a; proximity biotinylation, Co-IP, and double morpholino knockdown in Xenopus for SSX2IP","pmids":["34930905","34710136"],"confidence":"Medium","gaps":["Whether WTIP binds the FOXO3a promoter directly or through a cofactor is unresolved","SSX2IP–WTIP interaction confirmed only by proximity biotinylation/Co-IP without endogenous validation"]},{"year":2022,"claim":"Overexpression studies showed WTIP competes with LIMD1 at junctions and paradoxically promotes YAP phosphorylation (pro-Hippo effect), refining the model that WTIP cannot efficiently recruit Lats kinases like other Ajuba members, while domain mapping confirmed high-affinity WTIP–ARHGEF12 binding through LIM and PDZ domains. A hypertrophic cardiomyopathy–associated WTIP p.Y233F variant disrupts Wnt protein binding and alters cardiomyocyte calcium dynamics.","evidence":"Overexpression in MCF10A cells with phospho-YAP immunoblot; Co-IP domain mapping with heterozygous mouse injury model; shRNA in NRVMs, patient iPSC-CMs, and zebrafish morpholino for HCM variant","pmids":["36439396","35862649","35671065"],"confidence":"Medium","gaps":["WTIP-specific Hippo signaling output versus LIMD1 compensation not tested endogenously","Structural basis for failure to recruit Lats kinases not determined","HCM mechanism linking Y233F to calcium handling is incompletely defined"]},{"year":2025,"claim":"Discovery that WTIP recruits PP2A to inhibit AKT T308 phosphorylation established a new mechanism by which WTIP suppresses stemness and chemoresistance, with TET2/PATZ1 identified as upstream transcriptional activators of WTIP expression.","evidence":"Co-IP (WTIP–PP2A), AKT kinase assay, overexpression/knockdown, TET2/PATZ1 reporter assay in gastric cancer cells","pmids":["40279682"],"confidence":"Medium","gaps":["Single lab finding not yet independently replicated","Whether PP2A interaction occurs at junctions or in the cytosol is unclear","Relevance of TET2/PATZ1 axis to non-cancer WTIP biology not tested"]},{"year":null,"claim":"Major unresolved questions include: (1) how WTIP integrates its junctional, centrosomal, P-body, and nuclear functions in a single cell; (2) whether LIM domain conformational changes mediate tension sensing; (3) the structural basis distinguishing WTIP from LIMD1/Ajuba in Hippo signaling; and (4) whether WTIP mutations are causative for human kidney or cardiac disease.","evidence":"","pmids":[],"confidence":"Low","gaps":["No crystal or cryo-EM structure of WTIP or its complexes","No WTIP conditional knockout mouse phenotype reported","Disease causality for human WTIP variants not established by family segregation or rescue"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,5,7]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3,5,18]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[8,12]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,7]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[2,3,7]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[1,12]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[0]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[4,6,11,19]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,5,16,18]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[0]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[4,7,13]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[12]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[4,6,19]}],"complexes":["miRISC/P-body complex","Nephrin-Nck-WTIP-Lats1 complex"],"partners":["ARHGEF12","SHROOM3","LATS1","AGO2","ROR2","ASXL1","SSX2IP","PK3"],"other_free_text":[]},"mechanistic_narrative":"WTIP is an Ajuba-family LIM-domain scaffold protein that functions as a tension-sensitive adaptor at cell–cell junctions, coordinating actomyosin contractility, Hippo pathway signaling, miRNA-mediated gene silencing, and transcriptional regulation across multiple tissues including podocytes, neuroepithelium, and cardiomyocytes. At adherens junctions and focal adhesions, WTIP directly binds the RhoA-GEF ARHGEF12 to promote RhoA/ROCK-dependent stress fiber formation and interacts with Shroom3 to regulate apical constriction during neural tube closure [PMID:21900451, PMID:29661847, PMID:35862649]. WTIP suppresses Hippo signaling by linking nephrin/Nck complexes to Lats1, sequestering it from activation; upon injury-induced nephrin dephosphorylation, Lats1 is released and activated, leading to YAP downregulation [PMID:27033705, PMID:36439396]. WTIP also localizes to P-bodies where it bridges Ago1/2 to the eIF4E–m7GTP cap complex to enable miRNA-mediated translational silencing, undergoes JNK/dynein/microtubule-dependent nuclear translocation upon injury to modulate transcription factors including WT1 and FOXO3a, and inhibits AKT activity through PP2A recruitment [PMID:20616046, PMID:20086015, PMID:34930905, PMID:40279682]."},"prefetch_data":{"uniprot":{"accession":"A6NIX2","full_name":"Wilms tumor protein 1-interacting protein","aliases":[],"length_aa":430,"mass_kda":45.1,"function":"Adapter or scaffold protein which participates in the assembly of numerous protein complexes and is involved in several cellular processes such as cell fate determination, cytoskeletal organization, repression of gene transcription, cell-cell adhesion, cell differentiation, proliferation and migration. Positively regulates microRNA (miRNA)-mediated gene silencing. Negatively regulates Hippo signaling pathway and antagonizes phosphorylation of YAP1. Acts as a transcriptional corepressor for SNAI1 and SNAI2/SLUG-dependent repression of E-cadherin transcription. Acts as a hypoxic regulator by bridging an association between the prolyl hydroxylases and VHL enabling efficient degradation of HIF1A. In podocytes, may play a role in the regulation of actin dynamics and/or foot process cytoarchitecture (By similarity). In the course of podocyte injury, shuttles into the nucleus and acts as a transcription regulator that represses WT1-dependent transcription regulation, thereby translating changes in slit diaphragm structure into altered gene expression and a less differentiated phenotype. Involved in the organization of the basal body (By similarity). Involved in cilia growth and positioning (By similarity)","subcellular_location":"Cell junction, adherens junction; Nucleus; Cytoplasm, P-body","url":"https://www.uniprot.org/uniprotkb/A6NIX2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/WTIP","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/WTIP","total_profiled":1310},"omim":[{"mim_id":"614790","title":"WT1-INTERACTING PROTEIN; WTIP","url":"https://www.omim.org/entry/614790"},{"mim_id":"613026","title":"CHROMOSOME 19q13.11 DELETION SYNDROME, DISTAL","url":"https://www.omim.org/entry/613026"},{"mim_id":"612939","title":"HEAT-SHOCK 70-KD PROTEIN-BINDING PROTEIN 1; HSPBP1","url":"https://www.omim.org/entry/612939"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"blood vessel","ntpm":92.5}],"url":"https://www.proteinatlas.org/search/WTIP"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"A6NIX2","domains":[{"cath_id":"2.10.110.10","chopping":"366-416","consensus_level":"medium","plddt":89.1706,"start":366,"end":416}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/A6NIX2","model_url":"https://alphafold.ebi.ac.uk/files/AF-A6NIX2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-A6NIX2-F1-predicted_aligned_error_v6.png","plddt_mean":64.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=WTIP","jax_strain_url":"https://www.jax.org/strain/search?query=WTIP"},"sequence":{"accession":"A6NIX2","fasta_url":"https://rest.uniprot.org/uniprotkb/A6NIX2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/A6NIX2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/A6NIX2"}},"corpus_meta":[{"pmid":"20616046","id":"PMC_20616046","title":"LIM-domain proteins, LIMD1, Ajuba, and WTIP are required for microRNA-mediated gene silencing.","date":"2010","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/20616046","citation_count":53,"is_preprint":false},{"pmid":"22378287","id":"PMC_22378287","title":"19q13.11 cryptic deletion: description of two new cases and indication for a role of WTIP haploinsufficiency in hypospadias.","date":"2012","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/22378287","citation_count":43,"is_preprint":false},{"pmid":"23213452","id":"PMC_23213452","title":"Wtip and Vangl2 are required for mitotic spindle orientation and cloaca morphogenesis.","date":"2012","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/23213452","citation_count":32,"is_preprint":false},{"pmid":"27062996","id":"PMC_27062996","title":"Prickle3 synergizes with Wtip to regulate basal body organization and cilia growth.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27062996","citation_count":31,"is_preprint":false},{"pmid":"20086015","id":"PMC_20086015","title":"Podocyte injury induces nuclear translocation of WTIP via microtubule-dependent transport.","date":"2010","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20086015","citation_count":25,"is_preprint":false},{"pmid":"21900451","id":"PMC_21900451","title":"WT1-interacting protein (Wtip) regulates podocyte phenotype by cell-cell and cell-matrix contact reorganization.","date":"2011","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/21900451","citation_count":24,"is_preprint":false},{"pmid":"27033705","id":"PMC_27033705","title":"Nephrin Suppresses Hippo Signaling through the Adaptor Proteins Nck and WTIP.","date":"2016","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/27033705","citation_count":22,"is_preprint":false},{"pmid":"34930905","id":"PMC_34930905","title":"WTIP upregulates FOXO3a and induces apoptosis through PUMA in acute myeloid leukemia.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/34930905","citation_count":19,"is_preprint":false},{"pmid":"29661847","id":"PMC_29661847","title":"The Ajuba family protein Wtip regulates actomyosin contractility during vertebrate neural tube closure.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/29661847","citation_count":17,"is_preprint":false},{"pmid":"19785987","id":"PMC_19785987","title":"The LIM domain protein Wtip interacts with the receptor tyrosine kinase Ror2 and inhibits canonical Wnt signalling.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19785987","citation_count":16,"is_preprint":false},{"pmid":"34710136","id":"PMC_34710136","title":"Identification of the centrosomal maturation factor SSX2IP as a Wtip-binding partner by targeted proximity biotinylation.","date":"2021","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/34710136","citation_count":12,"is_preprint":false},{"pmid":"26385183","id":"PMC_26385183","title":"Role of Asxl1 in kidney podocyte development via its interaction with Wtip.","date":"2015","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/26385183","citation_count":11,"is_preprint":false},{"pmid":"25065743","id":"PMC_25065743","title":"WTIP interacts with ASXL2 and blocks ASXL2-mediated activation of retinoic acid signaling.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/25065743","citation_count":9,"is_preprint":false},{"pmid":"21686224","id":"PMC_21686224","title":"Out on a LIM: chronic kidney disease, podocyte phenotype and the Wilm's tumor interacting protein (WTIP).","date":"2011","source":"Transactions of the American Clinical and Climatological Association","url":"https://pubmed.ncbi.nlm.nih.gov/21686224","citation_count":9,"is_preprint":false},{"pmid":"24376653","id":"PMC_24376653","title":"Wtip- and gadd45a-interacting protein dendrin is not crucial for the development or maintenance of the glomerular filtration barrier.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24376653","citation_count":8,"is_preprint":false},{"pmid":"30179602","id":"PMC_30179602","title":"Identification of the UBA2-WTIP fusion gene in acute myeloid leukemia.","date":"2018","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/30179602","citation_count":5,"is_preprint":false},{"pmid":"27484451","id":"PMC_27484451","title":"Wtip is required for proepicardial organ specification and cardiac left/right asymmetry in zebrafish.","date":"2016","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/27484451","citation_count":5,"is_preprint":false},{"pmid":"35862649","id":"PMC_35862649","title":"Essential role of Wtip in mouse development and maintenance of the glomerular filtration barrier.","date":"2022","source":"American journal of physiology. Renal physiology","url":"https://pubmed.ncbi.nlm.nih.gov/35862649","citation_count":4,"is_preprint":false},{"pmid":"27535760","id":"PMC_27535760","title":"WTIP interacts with BRCA2 and is essential for BRCA2 centrosome localization in cervical cancer cell.","date":"2016","source":"Archives of gynecology and obstetrics","url":"https://pubmed.ncbi.nlm.nih.gov/27535760","citation_count":4,"is_preprint":false},{"pmid":"36439396","id":"PMC_36439396","title":"AJUBA and WTIP can compete with LIMD1 for junctional localization and LATS regulation.","date":"2022","source":"microPublication biology","url":"https://pubmed.ncbi.nlm.nih.gov/36439396","citation_count":3,"is_preprint":false},{"pmid":"35671065","id":"PMC_35671065","title":"Wnt Signaling Interactor WTIP (Wilms Tumor Interacting Protein) Underlies Novel Mechanism for Cardiac Hypertrophy.","date":"2022","source":"Circulation. Genomic and precision medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35671065","citation_count":3,"is_preprint":false},{"pmid":"40279682","id":"PMC_40279682","title":"DNA-methylation eraser TET2 activates WTIP expression to suppress an AKT-dependent chemoresistance of gastric cancer.","date":"2025","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/40279682","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11477,"output_tokens":4672,"usd":0.052256},"stage2":{"model":"claude-opus-4-6","input_tokens":8166,"output_tokens":4106,"usd":0.21522},"total_usd":0.267476,"stage1_batch_id":"msgbatch_01BparvgRZ9S2LMidvaoo86K","stage2_batch_id":"msgbatch_01NkBfv9SVZWthhS7j6mU6G3","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2010,\n      \"finding\": \"WTIP (along with LIMD1 and Ajuba) localizes to processing bodies (P-bodies) and binds Ago1/2, RCK, Dcp2, and eIF4E in vivo; these LIM proteins are required for miRNA-mediated (but not siRNA-mediated) gene silencing by linking the translationally inhibited eIF4E-m7GTP-5'-cap to Ago1/2 within the miRISC complex, forming an inhibitory closed-loop complex.\",\n      \"method\": \"Co-immunoprecipitation, m7GTP cap-binding assay, P-body imaging, knockdown with reporter silencing assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, functional knockdown with specific mechanistic readout, replicated across three family members with orthogonal assays\",\n      \"pmids\": [\"20616046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Upon LPS-induced podocyte injury, WTIP translocates from cell contacts to the nucleus via a mechanism requiring JNK activity, assembly of a JNK-interacting protein 3/dynein scaffold complex, intact microtubule networks, and dynein motor activity; loss of WTIP from cell contacts alters podocyte morphology and actin assembly.\",\n      \"method\": \"Live imaging, cellular fractionation, pharmacological inhibition (JNK inhibitor, microtubule disruption, dynein inhibition), shRNA knockdown with actin spreading assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (imaging, fractionation, pharmacological) in a single study with defined phenotypic readout\",\n      \"pmids\": [\"20086015\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"WTIP localizes to focal adhesions in isolated podocytes and shifts to adherens junctions upon homotypic cell contact; WTIP knockdown disrupts actin stress fiber formation, focal adhesion maturation, and adherens junction assembly. WTIP directly interacts with the RhoA-specific GEF ARHGEF12, and WTIP-induced stress fiber formation is blocked by RhoA inhibitor C3 toxin and ROCK inhibitor.\",\n      \"method\": \"shRNA knockdown, gain-of-function overexpression, cell surface biotinylation, co-immunoprecipitation, pharmacological inhibition (C3 toxin, ROCK inhibitor)\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, gain/loss-of-function with specific phenotypic readouts, pharmacological validation of pathway\",\n      \"pmids\": [\"21900451\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"WTIP physically interacts with the intracellular domain of the receptor tyrosine kinase Ror2; full-length Ror2 recruits WTIP to the cell membrane, whereas a disease-associated Ror2 mutant fails to do so. WTIP inhibits canonical Wnt signaling in mammalian cells and Xenopus embryos.\",\n      \"method\": \"Co-immunoprecipitation, cellular localization assay, Xenopus Wnt reporter assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single Co-IP with functional Wnt inhibition assay, single lab\",\n      \"pmids\": [\"19785987\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"In zebrafish, Wtip knockdown causes mitotic spindle orientation defects in the pronephros, cloaca malformation with reduced ciliated cells, and loss of striated rootlets from basal bodies resulting in absent cilia motility; Wtip and Vangl2 interact genetically (double morphant phenotype), placing Wtip in the planar cell polarity pathway.\",\n      \"method\": \"Morpholino knockdown in zebrafish, immunofluorescence, genetic epistasis (double morphant), high-speed video microscopy\",\n      \"journal\": \"Biology open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis plus multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"23213452\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Nephrin tyrosine phosphorylation recruits Nck adaptor proteins, which associate with WTIP and its binding partner Lats1, sequestering them to the nephrin complex and decreasing phospho-activation of Lats1; nephrin dephosphorylation (podocyte injury) leads to rapid Lats1 activation and subsequent YAP downregulation, connecting nephrin signaling to the Hippo pathway.\",\n      \"method\": \"Mutational analysis, co-immunoprecipitation, in vivo injury model (LPS mouse), immunoblot for phospho-Lats1/YAP\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — mutational analysis, reciprocal Co-IP, in vivo model with defined phosphorylation readouts\",\n      \"pmids\": [\"27033705\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Prickle3 (Pk3) physically associates with Wtip and cooperates with Wtip to regulate basal body organization and ciliogenesis in Xenopus GRP; Pk3 depletion prevents γ-tubulin and Nedd1 from associating with the basal body, and Wtip interaction is required for this Pk3 function.\",\n      \"method\": \"Co-immunoprecipitation, morpholino knockdown, immunofluorescence (γ-tubulin/Nedd1 localization), Xenopus GRP assay\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus functional epistasis in Xenopus, single lab\",\n      \"pmids\": [\"27062996\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Wtip localizes to apical cell junctions in Xenopus neuroectoderm and adopts a tension-sensitive distribution (punctate vs. linear) dependent on myosin II activity; depletion of Wtip inhibits apical constriction in neuroepithelial cells causing neural tube defects. The C-terminal LIM domain fragment of Wtip physically associates with Shroom3 and inhibits Shroom3-induced apical constriction.\",\n      \"method\": \"Morpholino knockdown in Xenopus, fluorescent protein tagging/live imaging, co-immunoprecipitation, dominant-negative C-terminal fragment assay\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — live imaging of tension-dependent localization, Co-IP, loss-of-function with defined morphogenetic phenotype\",\n      \"pmids\": [\"29661847\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"WTIP directly interacts with ASXL2 and represses ASXL2-mediated transcriptional activation of retinoic acid-responsive genes; WTIP blocks ASXL2's stimulatory effect on retinoic acid signaling in luciferase reporter assays.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, luciferase reporter assay in HeLa cells\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — Co-IP plus functional reporter assay, single lab\",\n      \"pmids\": [\"25065743\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"WTIP interacts with ASXL1; Asxl1-null mouse embryos show defects in kidney size and glomerular podocyte morphology along with upregulation of Wt1/Wtip target genes, implicating the Asxl1-Wtip interaction in maintenance of podocyte structure and regulation of WT1 signaling.\",\n      \"method\": \"Co-immunoprecipitation, Asxl1 knockout mouse, gene expression analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus KO mouse with defined phenotypic readout, single lab\",\n      \"pmids\": [\"26385183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Dendrin interacts with WTIP (and Gadd45a) at the podocyte slit diaphragm, as shown by yeast two-hybrid and co-immunoprecipitation; dendrin relocalizes to the nucleus in adriamycin nephropathy, suggesting a nuclear function linked to Wtip.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, immunofluorescence in injury model\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — yeast two-hybrid plus Co-IP, single lab\",\n      \"pmids\": [\"24376653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"WTIP interacts with BRCA2 (amino acids 2750–2864) in HeLa cells; WTIP knockdown abolishes BRCA2 centrosome localization and causes abnormal cell division.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, GST pull-down, shRNA knockdown, immunofluorescence microscopy\",\n      \"journal\": \"Archives of gynecology and obstetrics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — GST pull-down plus Co-IP plus KD phenotype, single lab\",\n      \"pmids\": [\"27535760\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"WTIP interacts with FOXO3a, transcriptionally activates FOXO3a expression, and promotes its nuclear translocation; this induces PUMA expression and activates the intrinsic apoptotic pathway, suppressing AML cell growth in vitro and in vivo.\",\n      \"method\": \"Co-immunoprecipitation, luciferase reporter (FOXO3a transcriptional activation), overexpression/knockdown, in vivo xenograft\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus transcriptional reporter plus in vivo model, single lab\",\n      \"pmids\": [\"34930905\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The N-terminal domain of Wtip physically associates with SSX2IP (a centriolar satellite/centrosomal maturation factor); double depletion of Wtip and SSX2IP in Xenopus embryos uncovered functional interaction required for neural tube closure and cell junction remodeling.\",\n      \"method\": \"Targeted proximity biotinylation, co-immunoprecipitation, co-localization imaging, double morpholino knockdown in Xenopus\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — proximity biotinylation followed by Co-IP and genetic epistasis, single lab\",\n      \"pmids\": [\"34710136\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"A UBA2-WTIP intrachromosomal fusion gene is present in AML; the fusion protein abrogates WTIP-mediated processing body (P-body) formation and induces phosphorylation of STAT3, STAT5, and ERK1/2, promoting cell proliferation.\",\n      \"method\": \"RT-PCR/Sanger sequencing, ectopic expression in KG-1a cells, immunofluorescence for P-bodies, immunoblot for pathway activation, in vivo xenograft\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional assays (P-body, signaling, proliferation) with in vivo validation, single lab\",\n      \"pmids\": [\"30179602\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"WTIP directly interacts with ARHGEF12 (a RhoA-specific GEF) through LIM- and PDZ-binding domains with high affinity; this interaction is proposed to regulate slit diaphragm and foot process function via Rho effector signaling.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping, heterozygous mouse model with LPS/adriamycin injury\",\n      \"journal\": \"American journal of physiology. Renal physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP with domain mapping plus in vivo injury model, single lab\",\n      \"pmids\": [\"35862649\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"WTIP overexpression in MCF10A cells reduces junctional localization of LIMD1 and LATS1, and is associated with increased YAP1 phosphorylation and decreased YAP1 nuclear localization, indicating WTIP competes with LIMD1 for adherens junction binding but cannot efficiently recruit Lats kinases, resulting in distinct (pro-Hippo) downstream effects.\",\n      \"method\": \"Overexpression in MCF10A cells, immunofluorescence for junctional localization, immunoblot for phospho-YAP1/YAP1 nuclear levels\",\n      \"journal\": \"microPublication biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment with functional Hippo pathway readout, single lab\",\n      \"pmids\": [\"36439396\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"WTIP knockdown in neonatal rat ventricular myocytes causes cellular hypertrophy; a HCM-associated variant WTIP p.Y233F disrupts binding to a Wnt signaling protein and alters cardiomyocyte calcium dynamics (increased resting and peak calcium), which is rescued by verapamil.\",\n      \"method\": \"shRNA knockdown in NRVMs, iPSC-derived cardiomyocytes from patient, calcium imaging, morpholino knockdown in zebrafish\",\n      \"journal\": \"Circulation. Genomic and precision medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple model systems (NRVMs, iPSC-CMs, zebrafish) with defined mechanistic readout, single lab\",\n      \"pmids\": [\"35671065\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"WTIP interacts with PP2A phosphatase and inhibits T308 phosphorylation and kinase activity of AKT, thereby suppressing stemness and chemoresistance in gastric cancer; TET2 and transcription factor PATZ1 cooperate to transcriptionally activate WTIP expression.\",\n      \"method\": \"Co-immunoprecipitation (WTIP-PP2A), AKT kinase assay (T308 phosphorylation), overexpression/knockdown, TET2/PATZ1 reporter assay\",\n      \"journal\": \"Neoplasia (New York, N.Y.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP plus kinase activity assay with functional chemoresistance readout, single lab\",\n      \"pmids\": [\"40279682\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In zebrafish, Wtip protein localizes to the basal body of proepicardial organ (PE) cells; wtip knockdown disrupts PE specification (in conjunction with WT1 signaling), perturbs cardiac looping, and disrupts early left-right asymmetry.\",\n      \"method\": \"Morpholino knockdown in zebrafish, mRNA overexpression, immunofluorescence for basal body localization, marker gene expression\",\n      \"journal\": \"Molecular medicine reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization experiment with loss-of-function phenotypic readout, single lab\",\n      \"pmids\": [\"27484451\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"WTIP is a LIM-domain scaffold protein of the Ajuba family that functions as a tension-sensitive adaptor at cell junctions, regulating actin/actomyosin contractility through interaction with ARHGEF12 (a RhoA-GEF) and Shroom3; it undergoes JNK/dynein/microtubule-dependent nuclear translocation in response to injury to modulate WT1 and FOXO3a transcription; it suppresses Hippo signaling by linking nephrin/Nck to Lats1, modulates miRNA-mediated gene silencing via Ago1/2 and the eIF4E-m7GTP cap complex in P-bodies, interacts with Ror2 to inhibit canonical Wnt signaling, and inhibits AKT activity through PP2A, collectively coordinating cell junction integrity, cytoskeletal dynamics, and transcriptional responses in podocytes, neuroepithelium, and other tissues.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"WTIP is an Ajuba-family LIM-domain scaffold protein that functions as a tension-sensitive adaptor at cell–cell junctions, coordinating actomyosin contractility, Hippo pathway signaling, miRNA-mediated gene silencing, and transcriptional regulation across multiple tissues including podocytes, neuroepithelium, and cardiomyocytes. At adherens junctions and focal adhesions, WTIP directly binds the RhoA-GEF ARHGEF12 to promote RhoA/ROCK-dependent stress fiber formation and interacts with Shroom3 to regulate apical constriction during neural tube closure [PMID:21900451, PMID:29661847, PMID:35862649]. WTIP suppresses Hippo signaling by linking nephrin/Nck complexes to Lats1, sequestering it from activation; upon injury-induced nephrin dephosphorylation, Lats1 is released and activated, leading to YAP downregulation [PMID:27033705, PMID:36439396]. WTIP also localizes to P-bodies where it bridges Ago1/2 to the eIF4E–m7GTP cap complex to enable miRNA-mediated translational silencing, undergoes JNK/dynein/microtubule-dependent nuclear translocation upon injury to modulate transcription factors including WT1 and FOXO3a, and inhibits AKT activity through PP2A recruitment [PMID:20616046, PMID:20086015, PMID:34930905, PMID:40279682].\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Establishing that WTIP is a Ror2-interacting protein that inhibits canonical Wnt signaling resolved the question of whether LIM-domain proteins at cell junctions could participate in Wnt pathway modulation.\",\n      \"evidence\": \"Co-immunoprecipitation and Xenopus Wnt reporter assay showing WTIP–Ror2 interaction and functional Wnt inhibition\",\n      \"pmids\": [\"19785987\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single Co-IP without reciprocal validation\", \"Endogenous interaction not confirmed\", \"Downstream targets of WTIP-mediated Wnt inhibition not identified\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Two contemporaneous studies established dual compartmentalization of WTIP: it localizes to P-bodies where it bridges Ago1/2 to the eIF4E–m7GTP cap for miRNA silencing, and it undergoes JNK/dynein/microtubule-dependent nuclear translocation from cell contacts upon injury, revealing stimulus-dependent shuttling as a core WTIP mechanism.\",\n      \"evidence\": \"Co-IP with cap-binding assays and knockdown reporter silencing for P-body function; live imaging, subcellular fractionation, and pharmacological inhibition for nuclear translocation\",\n      \"pmids\": [\"20616046\", \"20086015\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether P-body function and nuclear translocation are independently regulated is unclear\", \"Direct RNA targets of WTIP-dependent miRNA silencing not identified\", \"Nuclear transcriptional targets upon injury-induced translocation not fully defined\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identification of ARHGEF12 as a direct WTIP partner established the mechanism by which WTIP promotes RhoA/ROCK-dependent actin stress fiber formation at focal adhesions and adherens junctions, answering how a LIM scaffold controls actomyosin contractility.\",\n      \"evidence\": \"Co-IP, shRNA knockdown, overexpression, and pharmacological inhibition with C3 toxin and ROCK inhibitor in podocytes\",\n      \"pmids\": [\"21900451\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of WTIP–ARHGEF12 interaction not resolved\", \"Whether WTIP activates or scaffolds ARHGEF12 catalytic activity is unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Zebrafish studies revealed WTIP functions in planar cell polarity, controlling mitotic spindle orientation and ciliogenesis through genetic interaction with Vangl2, extending WTIP's role beyond cell junctions to basal body/cilium biology.\",\n      \"evidence\": \"Morpholino knockdown in zebrafish, double morphant epistasis with Vangl2, high-speed video microscopy of cilia motility\",\n      \"pmids\": [\"23213452\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Morpholino-only approach without genetic mutant confirmation\", \"Direct biochemical connection between WTIP and Vangl2 not established\", \"Mechanism of WTIP at the basal body not defined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovery that dendrin interacts with WTIP at the podocyte slit diaphragm and relocalizes to the nucleus upon injury added another injury-responsive WTIP partner, suggesting coordinated nuclear signaling from the slit diaphragm complex.\",\n      \"evidence\": \"Yeast two-hybrid and co-immunoprecipitation, immunofluorescence in adriamycin nephropathy model\",\n      \"pmids\": [\"24376653\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of dendrin–WTIP interaction not tested\", \"No reciprocal Co-IP reported\", \"Whether dendrin and WTIP co-translocate to the nucleus is unknown\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstration that WTIP represses ASXL2-mediated retinoic acid signaling expanded WTIP's nuclear functions to include transcriptional corepression of chromatin regulatory complexes.\",\n      \"evidence\": \"Yeast two-hybrid, co-immunoprecipitation, luciferase reporter assay in HeLa cells\",\n      \"pmids\": [\"25065743\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single Co-IP without domain mapping for ASXL2 interaction\", \"Endogenous relevance in podocytes or other tissues not tested\", \"Mechanism of repression (recruitment of corepressors?) not defined\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Asxl1 knockout mice showing kidney and podocyte defects with upregulated Wt1/Wtip targets provided in vivo genetic evidence that ASXL1–WTIP interaction maintains podocyte structure.\",\n      \"evidence\": \"Co-immunoprecipitation, Asxl1 knockout mouse, gene expression analysis\",\n      \"pmids\": [\"26385183\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"WTIP was not directly knocked out in this model\", \"Whether WTIP is the mediating partner versus other ASXL1 targets is unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Three parallel discoveries established WTIP as a node connecting nephrin signaling to Hippo pathway suppression (via Lats1 sequestration), cooperating with Prickle3 for basal body organization, and localizing to basal bodies in the proepicardial organ for cardiac left-right patterning.\",\n      \"evidence\": \"Nephrin mutational analysis with Co-IP and in vivo LPS model for Hippo; Pk3 Co-IP and Xenopus GRP morpholino epistasis for ciliogenesis; zebrafish morpholino KD with immunofluorescence for cardiac laterality\",\n      \"pmids\": [\"27033705\", \"27062996\", \"27484451\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct measurement of WTIP effect on Lats1 kinase activity in vitro is lacking\", \"Whether Pk3–WTIP and nephrin–WTIP complexes overlap in vivo is unknown\", \"Role of WTIP in proepicardial specification versus cilia function not separated\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of WTIP–BRCA2 interaction and requirement of WTIP for BRCA2 centrosome localization revealed an unexpected role in centrosome function and mitotic fidelity.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, GST pull-down, shRNA knockdown with immunofluorescence in HeLa cells\",\n      \"pmids\": [\"27535760\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab finding not independently replicated\", \"Whether WTIP scaffolds BRCA2 at centrosomes or regulates its transport is unclear\", \"Relationship to WTIP's basal body localization not explored\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Live imaging in Xenopus showed WTIP adopts tension-sensitive distributions at apical junctions dependent on myosin II and physically associates with Shroom3 to regulate apical constriction during neural tube closure, establishing WTIP as a mechanosensitive regulator of morphogenesis.\",\n      \"evidence\": \"Morpholino knockdown in Xenopus, fluorescent protein tagging/live imaging, Co-IP, dominant-negative C-terminal fragment assay\",\n      \"pmids\": [\"29661847\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether tension sensing involves conformational unfolding of LIM domains is not tested\", \"How WTIP coordinates Shroom3 and ARHGEF12 simultaneously is unresolved\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Discovery of a UBA2-WTIP fusion in AML that abrogates P-body formation and activates STAT3/5 and ERK1/2 demonstrated pathological consequences of disrupting WTIP's scaffold functions, linking WTIP loss-of-function to oncogenic signaling.\",\n      \"evidence\": \"RT-PCR/Sanger sequencing, ectopic expression in KG-1a cells, immunofluorescence for P-bodies, immunoblot, xenograft\",\n      \"pmids\": [\"30179602\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single patient fusion; frequency in AML cohorts not established\", \"Whether STAT/ERK activation is direct or secondary to P-body loss is unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Two studies expanded WTIP's functional network: WTIP transcriptionally activates FOXO3a to induce PUMA-mediated apoptosis in AML, and the N-terminus of WTIP associates with centriolar satellite protein SSX2IP for neural tube closure, revealing both nuclear transcriptional and centrosomal functions.\",\n      \"evidence\": \"Co-IP plus luciferase reporter plus in vivo xenograft for FOXO3a; proximity biotinylation, Co-IP, and double morpholino knockdown in Xenopus for SSX2IP\",\n      \"pmids\": [\"34930905\", \"34710136\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether WTIP binds the FOXO3a promoter directly or through a cofactor is unresolved\", \"SSX2IP–WTIP interaction confirmed only by proximity biotinylation/Co-IP without endogenous validation\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Overexpression studies showed WTIP competes with LIMD1 at junctions and paradoxically promotes YAP phosphorylation (pro-Hippo effect), refining the model that WTIP cannot efficiently recruit Lats kinases like other Ajuba members, while domain mapping confirmed high-affinity WTIP–ARHGEF12 binding through LIM and PDZ domains. A hypertrophic cardiomyopathy–associated WTIP p.Y233F variant disrupts Wnt protein binding and alters cardiomyocyte calcium dynamics.\",\n      \"evidence\": \"Overexpression in MCF10A cells with phospho-YAP immunoblot; Co-IP domain mapping with heterozygous mouse injury model; shRNA in NRVMs, patient iPSC-CMs, and zebrafish morpholino for HCM variant\",\n      \"pmids\": [\"36439396\", \"35862649\", \"35671065\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"WTIP-specific Hippo signaling output versus LIMD1 compensation not tested endogenously\", \"Structural basis for failure to recruit Lats kinases not determined\", \"HCM mechanism linking Y233F to calcium handling is incompletely defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Discovery that WTIP recruits PP2A to inhibit AKT T308 phosphorylation established a new mechanism by which WTIP suppresses stemness and chemoresistance, with TET2/PATZ1 identified as upstream transcriptional activators of WTIP expression.\",\n      \"evidence\": \"Co-IP (WTIP–PP2A), AKT kinase assay, overexpression/knockdown, TET2/PATZ1 reporter assay in gastric cancer cells\",\n      \"pmids\": [\"40279682\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab finding not yet independently replicated\", \"Whether PP2A interaction occurs at junctions or in the cytosol is unclear\", \"Relevance of TET2/PATZ1 axis to non-cancer WTIP biology not tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Major unresolved questions include: (1) how WTIP integrates its junctional, centrosomal, P-body, and nuclear functions in a single cell; (2) whether LIM domain conformational changes mediate tension sensing; (3) the structural basis distinguishing WTIP from LIMD1/Ajuba in Hippo signaling; and (4) whether WTIP mutations are causative for human kidney or cardiac disease.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No crystal or cryo-EM structure of WTIP or its complexes\", \"No WTIP conditional knockout mouse phenotype reported\", \"Disease causality for human WTIP variants not established by family segregation or rescue\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 5, 7]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3, 5, 18]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [8, 12]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [2, 3, 7]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [1, 12]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [4, 6, 11, 19]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 5, 16, 18]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [4, 7, 13]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [12]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [4, 6, 19]}\n    ],\n    \"complexes\": [\n      \"miRISC/P-body complex\",\n      \"Nephrin-Nck-WTIP-Lats1 complex\"\n    ],\n    \"partners\": [\n      \"ARHGEF12\",\n      \"SHROOM3\",\n      \"LATS1\",\n      \"AGO2\",\n      \"ROR2\",\n      \"ASXL1\",\n      \"SSX2IP\",\n      \"PK3\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}