{"gene":"ARHGAP24","run_date":"2026-06-09T22:02:44","timeline":{"discoveries":[{"year":2006,"finding":"FilGAP (ARHGAP24) is a Rac-specific GTPase-activating protein (GAP) that binds filamin A (FLNa). FLNa targets FilGAP to sites of membrane protrusion where it antagonizes Rac in vivo. ROCK phosphorylates FilGAP, and this phosphorylation stimulates its RacGAP activity and is required for FilGAP-mediated bleb formation. Knockdown of FilGAP abrogates ROCK-dependent suppression of lamellae, establishing FilGAP as a mediator of RhoA-Rac antagonism.","method":"siRNA knockdown, dominant-negative constructs, co-immunoprecipitation, in vitro kinase assay (ROCK phosphorylation), cell spreading assay on fibronectin, live-cell imaging of blebs","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal methods (biochemical binding, kinase assay, siRNA KD, dominant-negative), foundational paper replicated by subsequent work","pmids":["16862148"],"is_preprint":false},{"year":2009,"finding":"FilGAP interacts with the 23rd Ig repeat of filamin A (IgFLNa23) via the C-terminal 32 residues of FilGAP engaging the C and D beta-strands of IgFLNa23. Tight complex requires dimerization of both partners and a flexible hinge between FLNa repeats 23-24. FilGAP does not bind FLNa homologs FLNb or FLNc. Disease-associated FLNa mutations disrupt this binding interface, perturb cell spreading, and weaken elasticity of the FLNa/F-actin network under mechanical stress.","method":"NMR structural analysis, in silico modeling, mutagenesis of interface residues, cell spreading assay, F-actin network mechanics","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 1 / Moderate — NMR structure combined with mutagenesis and functional validation, single lab but multiple orthogonal methods","pmids":["19293932"],"is_preprint":false},{"year":2009,"finding":"FilGAP is targeted to sites of force transfer by FLNa (specifically repeat 23). Force-induced redistribution of FilGAP is essential for suppression of Rac activity and lamellae formation in cells treated with tensile forces applied through integrins. Depletion of FilGAP by siRNA, inhibition by dominant-negative mutation, or deletion of its FLNa-binding domain all result in dramatic force-induced increase of annexin-V-positive (apoptotic) cells, establishing FilGAP as a mechanoprotective anti-apoptotic effector downstream of FLNa.","method":"siRNA knockdown, dominant-negative constructs, integrin-mediated force application, annexin-V staining for apoptosis, fluorescence microscopy","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KD/KO with defined cellular phenotype plus dominant-negative and deletion constructs, single lab","pmids":["19144823"],"is_preprint":false},{"year":2011,"finding":"Mechanical strain in reconstituted actin/FLNa networks differentially regulates binding of FLNa partners: strain increases β-integrin binding to FLNa while causing FilGAP to dissociate from FLNa. Both externally imposed bulk shear and myosin-II-driven forces were shown to regulate this differential binding using fluorescence loss after photoconversion (FLAP).","method":"Reconstituted minimal system (actin filaments + FLNa + β-integrin tail + FilGAP), fluorescence loss after photoconversion (FLAP), bulk shear application, myosin-II inhibition","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with purified components, novel biophysical method, published in Nature with rigorous controls","pmids":["21926999"],"is_preprint":false},{"year":2011,"finding":"Arhgap24 is upregulated in differentiating kidney podocytes and functions as a RhoA-activated Rac1-GAP in these cells. Arhgap24 knockdown increases active Rac1 and Cdc42 levels and alters podocyte cell shape and membrane dynamics. A disease-associated missense mutation in ARHGAP24 impairs its Rac1-GAP activity and co-segregates with familial focal segmental glomerulosclerosis (FSGS).","method":"siRNA knockdown in cultured podocytes, GTPase activation assays (active Rac1/Cdc42 pulldown), Sanger sequencing of FSGS patients, in vitro GAP activity assay of mutant protein","journal":"The Journal of clinical investigation","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — in vitro GAP activity assay of mutant, cellular KD with GTPase measurements, human genetic validation, multiple orthogonal approaches","pmids":["21911940"],"is_preprint":false},{"year":2012,"finding":"FilGAP is a mediator of Rho/ROCK-dependent amoeboid movement of carcinoma cells. Depletion of FilGAP induces elongated mesenchymal morphology; forced expression induces round/amoeboid morphology requiring Rho/ROCK-dependent phosphorylation of FilGAP. Depletion of FilGAP impairs breast cancer cell invasion through ECM and reduces tumor cell extravasation in vivo.","method":"siRNA knockdown, forced expression, cell morphology analysis, Matrigel invasion assay, in vivo extravasation assay","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KD and OE with defined morphological and invasion phenotypes, in vivo validation, single lab","pmids":["23097497"],"is_preprint":false},{"year":2014,"finding":"FilGAP binds to activated Arf6 through its pleckstrin homology (PH) domain and is recruited to plasma membranes. Activated Arf6 stimulates the RacGAP activity of FilGAP. Knockdown of endogenous Arf6 by siRNA suppresses FilGAP-mediated bleb formation. A FilGAP mutant lacking PIP3 binding (R39C) still binds activated Arf6 and induces bleb formation, indicating a PIP3-independent interaction with Arf6.","method":"Co-immunoprecipitation, PH domain binding assay, siRNA knockdown, GAP activity assay, bleb formation assay, PI3K inhibitors","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — biochemical binding assay, GAP activity measurement, siRNA KD with functional readout, single lab multiple methods","pmids":["24526684"],"is_preprint":false},{"year":2015,"finding":"Phosphorylation of FilGAP serine 402 by ROCK regulates its subcellular localization: non-phosphorylatable FilGAP (S402A) localizes to actin cytoskeleton, while phosphomimetic or ROCK-phosphorylated FilGAP is diffusely cytoplasmic. Dephosphorylation of Ser-402 accompanies cell spreading on fibronectin. Calyculin A (Ser/Thr phosphatase inhibitor) suppresses spreading of WT FilGAP cells but not S402A cells. Arf6 activation stimulates bleb formation by both non-phosphorylatable and phosphomimetic FilGAP mutants.","method":"Site-directed mutagenesis (S402A, ST/A, ST/D), Phos-tag gel electrophoresis, fluorescence microscopy of localization, calyculin A treatment, cell spreading assay on fibronectin","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis with functional localization and spreading readouts, Phos-tag for phosphorylation state, single lab","pmids":["26359494"],"is_preprint":false},{"year":2015,"finding":"FilGAP promotes formation of adherens junctions in MDCK epithelial cells downstream of Rho-ROCK signaling. Knockdown of FilGAP stimulates HGF-induced disassembly and cell scattering; forced expression induces accumulation of E-cadherin at adherens junctions. The Rac GAP domain of FilGAP is required for suppression of HGF-induced scattering. ROCK-induced accumulation of E-cadherin at adherens junctions requires FilGAP and its ROCK-dependent phosphorylation (non-phosphorylatable FilGAP mutant fails to rescue).","method":"siRNA knockdown, forced expression, E-cadherin surface staining, HGF-induced scattering assay, ROCK overexpression, non-phosphorylatable mutant rescue","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean siRNA KD and OE with defined phenotypes, phospho-mutant analysis, single lab multiple orthogonal approaches","pmids":["25908853"],"is_preprint":false},{"year":2016,"finding":"RBM10 is a FilGAP-interacting protein. Src family tyrosine kinase Fyn induces translocation of RBM10 from nucleus to cell periphery in a kinase-activity-dependent manner, where RBM10 co-localizes with FilGAP. RBM10 is required for targeting FilGAP to the cell periphery during spreading, and association of RBM10 with FilGAP stimulates FilGAP's RacGAP activity as shown by suppression of spreading, ruffle formation, and FilGAP-dependent protrusive structures.","method":"Co-immunoprecipitation (FilGAP-RBM10), siRNA knockdown of RBM10, forced expression of Fyn (WT and kinase-dead), fluorescence co-localization, cell spreading assay, ruffle formation assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, siRNA KD with functional readout, kinase-dead mutant control, single lab","pmids":["26751795"],"is_preprint":false},{"year":2017,"finding":"ARHGAP24 is required for regulation of ARF6-dependent pseudopod formation in breast carcinoma cells. Knockdown of ARHGAP24 increases the lifespan and length of pseudopods induced by activated ARF6. ARHGAP24 requires its ARF6-binding site to achieve ARF6-dependent actin remodeling.","method":"siRNA knockdown of ARHGAP24, activated ARF6 expression, pseudopod measurement by time-lapse microscopy, binding-site mutant analysis","journal":"Anticancer research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, siRNA KD with morphological readout, limited mechanistic depth","pmids":["28870903"],"is_preprint":false},{"year":2019,"finding":"AGAP1 (an Arf GAP) binds to the C-terminus of FilGAP through AGAP1's N-terminal GLD domain. FilGAP co-localizes with AGAP1 at intracellular vesicles, and FilGAP targeting to these vesicles requires its interaction with AGAP1. Depletion of AGAP1 causes accumulation of endogenous FilGAP at paxillin-positive focal adhesions and actin structures, suppresses cell spreading on collagen (relieved by co-depletion of FilGAP), and promotes cell invasion that is blocked by FilGAP depletion.","method":"Co-immunoprecipitation, siRNA knockdown (AGAP1, FilGAP, double KD), fluorescence co-localization, cell spreading assay on collagen, Matrigel invasion assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain mapping, genetic epistasis via double KD with defined functional readouts, single lab","pmids":["31785816"],"is_preprint":false},{"year":2019,"finding":"FilGAP regulates distinct stages of MDCK epithelial tubulogenesis downstream of Rho-ROCK signaling. Depletion of FilGAP increases HGF-induced basolateral extensions from cysts; forced expression decreases extensions. Phosphomimetic FilGAP (ST/D) blocks membrane extension formation even in the presence of ROCK inhibitor Y-27632, and promotes lumen formation while blocking cell scattering in tubules.","method":"siRNA knockdown, forced expression, 3D MDCK cyst culture in ECM, HGF stimulation, ROCK inhibitor Y-27632, phosphomimetic mutant (ST/D)","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, KD/OE with morphological readout, limited mechanistic depth beyond Rho-ROCK axis","pmids":["31078260"],"is_preprint":false},{"year":2021,"finding":"FilGAP controls front-rear polarity and tumor cell migration speed through extracellular matrix. FilGAP localizes to the cell front through its PH domain in a PIP3-dependent manner to inactivate Rac at that site. Small GTPase Arf6 (binding to the FilGAP PH domain) also regulates FilGAP-mediated cell polarity and migration.","method":"Forced expression and siRNA depletion in breast cancer cells, collagen matrix migration assay, PH domain mutant analysis, PIP3 depletion, Arf6 manipulation","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mutant analysis combined with migration assay and localization studies, single lab, multiple methods","pmids":["33710706"],"is_preprint":false},{"year":2021,"finding":"CBX3 represses ARHGAP24 expression by forming a complex with TRIM28, TRIM24, and RBBP4, resulting in increased active Rac1 and promotion of lung adenocarcinoma progression.","method":"Co-immunoprecipitation of CBX3/TRIM28/TRIM24/RBBP4 complex, ChIP, Rac1 activity assay, ARHGAP24 expression measurement by western blot/qPCR","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP of multiprotein complex with functional validation of Rac1 activity, single lab","pmids":["34785774"],"is_preprint":false},{"year":2022,"finding":"ARHGAP24 acts as a GTPase-independent scaffold in hepatocellular carcinoma to suppress β-catenin signaling. It recruits the E3 ubiquitin ligase WWP1 to promote degradation of pyruvate kinase M2 (PKM2). The C-terminal fragment of ARHGAP24 (aa 329-430) binds WWP1 and aa 631-748 binds PKM2 (aa 388-531). A GAP-deficient ARHGAP24 mutant exerts the same inhibitory effects as wild-type, confirming GAP-independent scaffolding function.","method":"Liquid chromatography-tandem mass spectrometry, co-immunoprecipitation, ubiquitination assay, GAP activity assay (GAP-deficient mutant), luciferase reporter for β-catenin, domain mapping with deletion constructs","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mass spectrometry-based binding partner identification, co-IP with domain mapping, ubiquitination assay, GAP-dead mutant control, single lab","pmids":["36168627"],"is_preprint":false},{"year":2023,"finding":"FilGAP interacts with mTORC1 via Raptor and with mTORC2 via Rictor and Sin1. Depletion of FilGAP decreases phosphorylation of S6K (mTORC1 substrate) and AKT (mTORC2 substrate) in KINGS-1 glioma cells; conversely, overexpression of FilGAP increases these phosphorylations, indicating FilGAP activates mTORC1/2. FilGAP depletion reduces spheroid growth.","method":"Co-immunoprecipitation (FilGAP with Raptor/Rictor/Sin1), siRNA knockdown, forced expression, western blot for p-S6K and p-AKT, PI3K inhibitor treatment, spheroid growth assay","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP with mTOR complex components, KD and OE with biochemical readouts, single lab","pmids":["38065968"],"is_preprint":false},{"year":2023,"finding":"FilGAP localizes to invadopodia through its PH domain binding to phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2] and negatively regulates invadopodia formation by inactivating Rac1. Depletion of FilGAP or expression of GAP-deficient FilGAP mutant increases invadopodia formation and ECM degradation; overexpression suppresses both. PI(3,4)P2 depletion reduces FilGAP invadopodia localization.","method":"siRNA knockdown, forced expression, GAP-deficient mutant, PH domain PI(3,4)P2-binding mutant, gelatin degradation assay, EGF stimulation, fluorescence localization","journal":"Cell structure and function","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mutant analysis with functional and localization readouts, GAP-dead control, lipid-binding specificity tested, single lab","pmids":["37482421"],"is_preprint":false},{"year":2024,"finding":"FilGAP is phosphorylated at Ser625 by p90 ribosomal S6 kinase (RSK) downstream of EGF signaling, followed by phosphorylation at Ser621 by GSK3 in a sequential manner. This phosphorylation induces dissociation of FilGAP from actin filaments via a novel actin-localization domain. Phosphorylation inhibits FilGAP's lamellipodia-suppression activity. Non-phosphorylatable FilGAP mutant reduces cell migration speed and persistence toward EGF gradient, establishing this cascade as a regulator of chemotactic tumor cell migration.","method":"In vitro kinase assays (RSK, GSK3), site-directed mutagenesis (S625A, S621A, non-phosphorylatable mutant), actin co-sedimentation, EGF stimulation, chemotaxis assay, fluorescence localization","journal":"PNAS nexus","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay with site-specific mutagenesis, actin binding assay, functional chemotaxis readout, multiple orthogonal methods in single study","pmids":["38426123"],"is_preprint":false},{"year":2024,"finding":"FilGAP controls cell-ECM adhesion and process formation in podocytes by suppressing Rac1/PAK1 signaling. FilGAP localizes to focal adhesions (FAs); its depletion decreases FA formation, impairs attachment to ECM, increases Rac1 activity, and reduces formation of major processes and foot process-like projections. These deficits are rescued by inhibition of Rac1 or PAK1, placing PAK1 as a downstream effector of FilGAP/Rac1 in podocyte morphology.","method":"siRNA knockdown, forced expression, Rac1 activity pulldown assay, focal adhesion staining (paxillin/vinculin), podocyte process formation assay, Rac1 inhibitor, PAK1 inhibitor rescue","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis via inhibitor rescue, KD with multiple cellular readouts, single lab","pmids":["38421271"],"is_preprint":false},{"year":2024,"finding":"Loss-of-function variants in ARHGAP24 found in families with isolated posterior mitral valve prolapse (PostMVP) impair cellular adhesion and mechanotransduction capacities in vitro. Silencing of ARHGAP24 in zebrafish leads to atrioventricular regurgitation, establishing a functional role for FilGAP in cardiac valve development/maintenance.","method":"Zebrafish morpholino silencing (in vivo valve phenotype), in vitro functional assays of FilGAP variants (cell adhesion, mechanotransduction), whole genome sequencing of families","journal":"bioRxiv (preprint)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo zebrafish model with defined phenotype, in vitro functional validation of human variants, single study, preprint","pmids":["bio_10.1101_2024.10.16.24315096"],"is_preprint":true}],"current_model":"ARHGAP24/FilGAP is a Rac1-specific GTPase-activating protein that binds filamin A (via IgFLNa23) and is activated by ROCK-mediated phosphorylation (including at Ser402) downstream of RhoA, thereby mediating Rho-Rac antagonism to control actin remodeling, cell spreading, bleb formation, adherens junctions, focal adhesions, and cell polarity; mechanical strain on FLNa causes FilGAP to dissociate from FLNa (mechanotransduction); FilGAP is additionally regulated by Arf6 (binding through its PH domain to PIP3/PI(3,4)P2), by EGF-driven RSK/GSK3 phosphorylation at Ser625/621 (promoting actin dissociation and chemotactic migration), and by RBM10/Fyn-kinase signaling; it also acts as a GTPase-independent scaffold recruiting WWP1 to degrade PKM2 and suppress β-catenin signaling, and activates mTORC1/2 via interactions with Raptor, Rictor, and Sin1."},"narrative":{"mechanistic_narrative":"ARHGAP24 (FilGAP) is a Rac1-specific GTPase-activating protein that integrates RhoA/ROCK signaling with actin remodeling to enforce Rho-Rac antagonism during cell spreading, motility, and tissue morphogenesis [PMID:16862148, PMID:21911940]. It is targeted to sites of membrane protrusion and force transfer by binding the 23rd Ig repeat of filamin A through its C-terminal residues, an interaction requiring dimerization of both partners; mechanical strain on the actin/FLNa network causes FilGAP to dissociate, coupling mechanotransduction to local Rac inactivation and mechanoprotection against force-induced apoptosis [PMID:19293932, PMID:19144823, PMID:21926999]. ROCK phosphorylation, including at Ser402, stimulates its RacGAP activity and controls its partition between the actin cytoskeleton and cytoplasm, with dephosphorylation accompanying cell spreading on fibronectin [PMID:16862148, PMID:26359494]. FilGAP localization is further directed by phosphoinositides and Arf6: its PH domain binds PIP3/PI(3,4)P2 and activated Arf6 to recruit it to the plasma membrane, cell front, and invadopodia, where it locally inactivates Rac1 to govern bleb and pseudopod formation, front-rear polarity, and ECM invasion [PMID:24526684, PMID:33710706, PMID:37482421]. An EGF-driven RSK-then-GSK3 cascade phosphorylates Ser625/Ser621 to release FilGAP from actin and promote chemotactic migration, while RBM10/Fyn and AGAP1 control its peripheral and vesicular targeting [PMID:38426123, PMID:26751795, PMID:31785816]. Through these activities FilGAP shapes adherens junctions, focal adhesions, and epithelial tubulogenesis downstream of Rho-ROCK [PMID:25908853, PMID:38421271], and beyond its GAP function it acts as a scaffold that recruits WWP1 to degrade PKM2 and suppress β-catenin signaling and that engages Raptor, Rictor, and Sin1 to activate mTORC1/2 [PMID:36168627, PMID:38065968]. Loss of ARHGAP24 function causes familial focal segmental glomerulosclerosis through defective podocyte Rac1/PAK1 control [PMID:21911940, PMID:38421271].","teleology":[{"year":2006,"claim":"Established FilGAP as the molecular link that allows RhoA/ROCK signaling to suppress Rac-driven lamellae, defining a dedicated effector of Rho-Rac antagonism rather than treating the two GTPases as independently regulated.","evidence":"siRNA knockdown, dominant-negative constructs, co-IP with filamin A, in vitro ROCK kinase assay, and bleb/spreading imaging","pmids":["16862148"],"confidence":"High","gaps":["Did not resolve the structural basis of the FLNa interaction","Phosphosites mediating ROCK activation not yet mapped"]},{"year":2009,"claim":"Defined the structural interface tethering FilGAP to filamin A and showed disease FLNa mutations disrupt it, explaining how FilGAP is positioned at force-bearing actin sites.","evidence":"NMR structure of the IgFLNa23/FilGAP C-terminus complex, interface mutagenesis, and F-actin network mechanics; complemented by force-application studies showing force-induced redistribution is required to suppress Rac and protect against apoptosis","pmids":["19293932","19144823"],"confidence":"High","gaps":["Mechanism converting FLNa conformational change into GAP activation not detailed","Apoptosis link single-lab"]},{"year":2011,"claim":"Demonstrated directly that mechanical strain acts as a switch dissociating FilGAP from filamin A, providing a biophysical mechanism for mechanotransduction at the actin network.","evidence":"Reconstituted actin/FLNa/β-integrin/FilGAP system with FLAP under bulk shear and myosin-II-driven forces","pmids":["21926999"],"confidence":"High","gaps":["Quantitative force thresholds in cells not established","Downstream consequence of dissociation in vivo inferred"]},{"year":2011,"claim":"Connected ARHGAP24 to human disease and a physiological cell type, showing its Rac1-GAP activity maintains podocyte architecture and that a GAP-impairing mutation causes familial FSGS.","evidence":"Podocyte siRNA knockdown with active Rac1/Cdc42 pulldowns, Sanger sequencing of FSGS families, and in vitro GAP assay of mutant protein","pmids":["21911940"],"confidence":"High","gaps":["Did not connect podocyte function to downstream effector (later resolved as PAK1)","RhoA-activation mechanism in podocytes not detailed"]},{"year":2014,"claim":"Identified Arf6 as an activator and membrane-recruitment partner acting through the FilGAP PH domain, separating PIP3 binding from Arf6 binding as distinct regulatory inputs.","evidence":"Co-IP, PH-domain binding assays, GAP activity measurement, siRNA Arf6 depletion, and PIP3-binding mutant (R39C) bleb assays","pmids":["24526684"],"confidence":"Medium","gaps":["Structural basis of Arf6/PH interaction not resolved","Single lab"]},{"year":2015,"claim":"Showed ROCK phosphorylation of Ser402 acts as a localization switch between actin and cytoplasm, and that FilGAP promotes adherens junction assembly, extending its role to epithelial integrity.","evidence":"S402A/phosphomimetic mutants with Phos-tag and localization imaging; MDCK E-cadherin staining, HGF scattering, and phospho-mutant rescue","pmids":["26359494","25908853"],"confidence":"Medium","gaps":["Phosphatase responsible for Ser402 dephosphorylation not identified","Direct GAP target at junctions not measured"]},{"year":2016,"claim":"Revealed RBM10 as a Fyn-regulated partner that relocates from nucleus to periphery to target FilGAP and stimulate its RacGAP activity, adding a tyrosine-kinase-controlled targeting layer.","evidence":"Reciprocal co-IP, RBM10 siRNA, WT/kinase-dead Fyn expression, and spreading/ruffle assays","pmids":["26751795"],"confidence":"Medium","gaps":["Whether Fyn phosphorylates FilGAP directly not shown","Physiological context of RBM10 relocation unclear"]},{"year":2017,"claim":"Placed FilGAP downstream of Arf6 in controlling pseudopod dynamics in carcinoma cells, linking its Arf6-binding site to actin remodeling during invasion.","evidence":"ARHGAP24 siRNA with activated Arf6 expression, time-lapse pseudopod measurement, and binding-site mutant","pmids":["28870903"],"confidence":"Low","gaps":["Limited mechanistic depth, morphological readout only","Single lab, low confidence"]},{"year":2019,"claim":"Identified AGAP1 as a partner directing FilGAP to intracellular vesicles and away from focal adhesions, with epistasis defining FilGAP as the effector of AGAP1-controlled spreading and invasion.","evidence":"Co-IP with GLD-domain mapping, single and double siRNA knockdown, spreading on collagen, and Matrigel invasion; complemented by 3D tubulogenesis KD/OE studies","pmids":["31785816","31078260"],"confidence":"Medium","gaps":["How vesicular vs FA targeting is balanced not quantified","Tubulogenesis study low confidence"]},{"year":2021,"claim":"Defined FilGAP as a determinant of front-rear polarity, recruited to the cell front by PIP3 and Arf6 to locally inactivate Rac and tune migration speed, and showed its expression is epigenetically repressed in cancer.","evidence":"PH-domain mutant migration assays with PIP3/Arf6 manipulation in breast cancer cells; CBX3/TRIM28/TRIM24/RBBP4 complex co-IP, ChIP, and Rac1 activity in lung adenocarcinoma","pmids":["33710706","34785774"],"confidence":"Medium","gaps":["Spatial coordination of PIP3 and Arf6 inputs not resolved","CBX3 repression mechanism single lab"]},{"year":2022,"claim":"Uncovered a GAP-independent scaffolding function in which FilGAP bridges WWP1 to PKM2 for degradation and suppresses β-catenin signaling, expanding its role beyond Rac regulation.","evidence":"LC-MS/MS partner identification, co-IP with domain mapping, ubiquitination assay, β-catenin luciferase reporter, and GAP-deficient mutant control","pmids":["36168627"],"confidence":"Medium","gaps":["In vivo relevance to hepatocellular carcinoma not fully established","Single lab"]},{"year":2023,"claim":"Connected FilGAP to growth signaling and invadopodia, showing it activates mTORC1/2 via Raptor/Rictor/Sin1 while its PI(3,4)P2-driven invadopodial localization restrains Rac1-dependent ECM degradation.","evidence":"Co-IP with mTOR components plus p-S6K/p-AKT and spheroid readouts; invadopodia gelatin degradation with PH-domain and GAP-dead mutants and PI(3,4)P2 depletion","pmids":["38065968","37482421"],"confidence":"Medium","gaps":["Whether mTOR activation is GAP-dependent unresolved","Direct biochemical link between FilGAP and mTOR activation unclear"]},{"year":2024,"claim":"Mapped a sequential RSK-then-GSK3 phosphorylation cascade at Ser625/Ser621 that releases FilGAP from actin to drive EGF chemotaxis, and tied FilGAP/Rac1/PAK1 to podocyte adhesion and cardiac valve integrity.","evidence":"In vitro RSK/GSK3 kinase assays with site mutants and actin co-sedimentation plus chemotaxis; podocyte KD with Rac1/PAK1 inhibitor rescue; zebrafish silencing and human PostMVP variant functional assays (preprint)","pmids":["38426123","38421271","bio_10.1101_2024.10.16.24315096"],"confidence":"High","gaps":["Phosphatase reversing Ser625/621 not identified","PostMVP finding is a preprint with single zebrafish model"]},{"year":null,"claim":"How FilGAP's diverse regulatory inputs (FLNa mechanics, ROCK/RSK/GSK3 phosphorylation, Arf6 and phosphoinositide targeting) are integrated in real time, and how its GAP-dependent and GAP-independent scaffolding functions are partitioned, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unified structural model coupling phosphorylation, lipid, and FLNa binding to GAP activation","Switch between Rac-GAP and WWP1/PKM2 or mTOR scaffolding roles undefined","In vivo physiological hierarchy of regulatory inputs untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,4,6]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[6,13,17]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[15,16]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,18]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,13]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[7,18]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[7]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[11]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,4,13]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,15,20]},{"term_id":"R-HSA-1474244","term_label":"Extracellular matrix organization","supporting_discovery_ids":[5,17,19]}],"complexes":["mTORC1","mTORC2"],"partners":["FLNA","RBM10","AGAP1","WWP1","PKM2","RPTOR","RICTOR","MAPKAP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N264","full_name":"Rho GTPase-activating protein 24","aliases":["Filamin-A-associated RhoGAP","FilGAP","RAC1- and CDC42-specific GTPase-activating protein of 72 kDa","RC-GAP72","Rho-type GTPase-activating protein 24","RhoGAP of 73 kDa","Sarcoma antigen NY-SAR-88","p73RhoGAP"],"length_aa":748,"mass_kda":84.3,"function":"Rho GTPase-activating protein involved in cell polarity, cell morphology and cytoskeletal organization. Acts as a GTPase activator for the Rac-type GTPase by converting it to an inactive GDP-bound state. Controls actin remodeling by inactivating Rac downstream of Rho leading to suppress leading edge protrusion and promotes cell retraction to achieve cellular polarity. Able to suppress RAC1 and CDC42 activity in vitro. Overexpression induces cell rounding with partial or complete disruption of actin stress fibers and formation of membrane ruffles, lamellipodia, and filopodia. Isoform 2 is a vascular cell-specific GAP involved in modulation of angiogenesis","subcellular_location":"Cytoplasm, cytoskeleton; Cell junction, adherens junction; Cell junction, focal adhesion; Cell projection","url":"https://www.uniprot.org/uniprotkb/Q8N264/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/ARHGAP24","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/ARHGAP24","total_profiled":1310},"omim":[{"mim_id":"617485","title":"WD REPEAT- AND FYVE DOMAIN-CONTAINING PROTEIN 3; WDFY3","url":"https://www.omim.org/entry/617485"},{"mim_id":"610587","title":"RHO GTPase-ACTIVATING PROTEIN 25; ARHGAP25","url":"https://www.omim.org/entry/610587"},{"mim_id":"610586","title":"RHO GTPase-ACTIVATING PROTEIN 24; ARHGAP24","url":"https://www.omim.org/entry/610586"},{"mim_id":"610585","title":"RHO GTPase-ACTIVATING PROTEIN 22; ARHGAP22","url":"https://www.omim.org/entry/610585"},{"mim_id":"603278","title":"FOCAL SEGMENTAL GLOMERULOSCLEROSIS 1; FSGS1","url":"https://www.omim.org/entry/603278"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Plasma membrane","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"kidney","ntpm":93.2}],"url":"https://www.proteinatlas.org/search/ARHGAP24"},"hgnc":{"alias_symbol":["DKFZP564B1162","FLJ33877","FilGAP"],"prev_symbol":[]},"alphafold":{"accession":"Q8N264","domains":[{"cath_id":"2.30.29.30","chopping":"23-129","consensus_level":"high","plddt":88.1899,"start":23,"end":129},{"cath_id":"1.10.555.10","chopping":"138-329","consensus_level":"medium","plddt":93.5832,"start":138,"end":329}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N264","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N264-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N264-F1-predicted_aligned_error_v6.png","plddt_mean":65.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=ARHGAP24","jax_strain_url":"https://www.jax.org/strain/search?query=ARHGAP24"},"sequence":{"accession":"Q8N264","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N264.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N264/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N264"}},"corpus_meta":[{"pmid":"16862148","id":"PMC_16862148","title":"FilGAP, a Rho- and ROCK-regulated GAP for Rac binds filamin A to control actin remodelling.","date":"2006","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16862148","citation_count":309,"is_preprint":false},{"pmid":"21926999","id":"PMC_21926999","title":"Mechanical strain in actin networks regulates FilGAP and integrin binding to filamin A.","date":"2011","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/21926999","citation_count":278,"is_preprint":false},{"pmid":"21911940","id":"PMC_21911940","title":"Arhgap24 inactivates Rac1 in mouse podocytes, and a mutant form is associated with familial focal segmental glomerulosclerosis.","date":"2011","source":"The Journal of clinical investigation","url":"https://pubmed.ncbi.nlm.nih.gov/21911940","citation_count":213,"is_preprint":false},{"pmid":"23763313","id":"PMC_23763313","title":"FilGAP and its close relatives: a mediator of Rho-Rac antagonism that regulates cell morphology and migration.","date":"2013","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/23763313","citation_count":76,"is_preprint":false},{"pmid":"23097497","id":"PMC_23097497","title":"FilGAP, a Rho/Rho-associated protein kinase-regulated GTPase-activating protein for Rac, controls tumor cell migration.","date":"2012","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/23097497","citation_count":65,"is_preprint":false},{"pmid":"19144823","id":"PMC_19144823","title":"The role of FilGAP-filamin A interactions in mechanoprotection.","date":"2009","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/19144823","citation_count":65,"is_preprint":false},{"pmid":"19293932","id":"PMC_19293932","title":"Molecular basis of filamin A-FilGAP interaction and its impairment in congenital disorders associated with filamin A mutations.","date":"2009","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/19293932","citation_count":59,"is_preprint":false},{"pmid":"15254788","id":"PMC_15254788","title":"Identification and characterization of ARHGAP24 and ARHGAP25 genes in silico.","date":"2004","source":"International journal of molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/15254788","citation_count":51,"is_preprint":false},{"pmid":"31935390","id":"PMC_31935390","title":"HOTAIRM1 suppresses cell proliferation and invasion in ovarian cancer through facilitating ARHGAP24 expression by sponging miR-106a-5p.","date":"2020","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31935390","citation_count":44,"is_preprint":false},{"pmid":"27385097","id":"PMC_27385097","title":"ARHGAP24 inhibits cell cycle progression, induces apoptosis and suppresses invasion in renal cell carcinoma.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27385097","citation_count":39,"is_preprint":false},{"pmid":"31430374","id":"PMC_31430374","title":"ARHGAP24 inhibits cell proliferation and cell cycle progression and induces apoptosis of lung cancer via a STAT6-WWP2-p27 axis.","date":"2020","source":"Carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/31430374","citation_count":37,"is_preprint":false},{"pmid":"34785774","id":"PMC_34785774","title":"Smoking-associated upregulation of CBX3 suppresses ARHGAP24 expression to activate Rac1 signaling and promote tumor progression in lung adenocarcinoma.","date":"2021","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/34785774","citation_count":29,"is_preprint":false},{"pmid":"26751795","id":"PMC_26751795","title":"Src Family Tyrosine Kinase Signaling Regulates FilGAP through Association with RBM10.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26751795","citation_count":25,"is_preprint":false},{"pmid":"25908853","id":"PMC_25908853","title":"FilGAP, a Rho-ROCK-regulated GAP for Rac, controls adherens junctions in MDCK cells.","date":"2015","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/25908853","citation_count":21,"is_preprint":false},{"pmid":"24526684","id":"PMC_24526684","title":"ADP ribosylation factor 6 (Arf6) acts through FilGAP protein to down-regulate Rac protein and regulates plasma membrane blebbing.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24526684","citation_count":19,"is_preprint":false},{"pmid":"30127956","id":"PMC_30127956","title":"ARHGAP24 regulates cell ability and apoptosis of colorectal cancer cells via the regulation of P53.","date":"2018","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/30127956","citation_count":18,"is_preprint":false},{"pmid":"29019371","id":"PMC_29019371","title":"MicroRNA-590-5p regulates cell viability, apoptosis, migration and invasion of renal cell carcinoma cell lines through targeting ARHGAP24.","date":"2017","source":"Molecular bioSystems","url":"https://pubmed.ncbi.nlm.nih.gov/29019371","citation_count":18,"is_preprint":false},{"pmid":"26359494","id":"PMC_26359494","title":"Phosphorylation of Serine 402 Regulates RacGAP Protein Activity of FilGAP Protein.","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26359494","citation_count":17,"is_preprint":false},{"pmid":"36168627","id":"PMC_36168627","title":"ARHGAP24 represses β-catenin transactivation-induced invasiveness in hepatocellular carcinoma mainly by acting as a GTPase-independent scaffold.","date":"2022","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/36168627","citation_count":16,"is_preprint":false},{"pmid":"27790861","id":"PMC_27790861","title":"The role of FilGAP, a Rac-specific Rho-GTPase-activating protein, in tumor progression and behavior of astrocytomas.","date":"2016","source":"Cancer medicine","url":"https://pubmed.ncbi.nlm.nih.gov/27790861","citation_count":14,"is_preprint":false},{"pmid":"35713208","id":"PMC_35713208","title":"Role of miR-21-5p/FilGAP axis in estradiol alleviating the progression of monocrotaline-induced pulmonary hypertension.","date":"2022","source":"Animal models and experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35713208","citation_count":14,"is_preprint":false},{"pmid":"38742566","id":"PMC_38742566","title":"Exosome-transmitted circular RNA circ-LMO7 facilitates the progression of osteosarcoma by regulating miR-21-5p/ARHGAP24 axis.","date":"2024","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/38742566","citation_count":14,"is_preprint":false},{"pmid":"30599132","id":"PMC_30599132","title":"Rho GTPase Activating Protein 24 (ARHGAP24) Silencing Promotes Lung Cancer Cell Migration and Invasion by Activating β-Catenin Signaling.","date":"2019","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/30599132","citation_count":14,"is_preprint":false},{"pmid":"31785816","id":"PMC_31785816","title":"AGAP1 regulates subcellular localization of FilGAP and control cancer cell invasion.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31785816","citation_count":13,"is_preprint":false},{"pmid":"33209869","id":"PMC_33209869","title":"ARHGAP24 ameliorates inflammatory response through inactivating Rac1/Akt/NF-κB pathway in acute pneumonia model of rat.","date":"2020","source":"Annals of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33209869","citation_count":13,"is_preprint":false},{"pmid":"33710706","id":"PMC_33710706","title":"FilGAP, a GAP protein for Rac, regulates front-rear polarity and tumor cell migration through the ECM.","date":"2021","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/33710706","citation_count":12,"is_preprint":false},{"pmid":"30499465","id":"PMC_30499465","title":"Rho GTPase Activating Protein 24 (ARHGAP24) Regulates the Anti-Cancer Activity of Sorafenib Against Breast Cancer MDA-MB-231 Cells via the Signal Transducer and Activator of Transcription 3 (STAT3) Signaling Pathway.","date":"2018","source":"Medical science monitor : international medical journal of experimental and clinical research","url":"https://pubmed.ncbi.nlm.nih.gov/30499465","citation_count":11,"is_preprint":false},{"pmid":"28870903","id":"PMC_28870903","title":"Role of ARHGAP24 in ADP Ribosylation Factor 6 (ARF6)-dependent Pseudopod Formation in Human Breast Carcinoma Cells.","date":"2017","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/28870903","citation_count":10,"is_preprint":false},{"pmid":"35812178","id":"PMC_35812178","title":"RELA-induced MiR-21 Exerts Oncogenic Effects on PDAC via Targeting of ARHGAP24.","date":"2022","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35812178","citation_count":8,"is_preprint":false},{"pmid":"35112252","id":"PMC_35112252","title":"miR-21-5p serves as a promoter in renal cell carcinoma progression through ARHGAP24 downregulation.","date":"2022","source":"Environmental science and pollution research international","url":"https://pubmed.ncbi.nlm.nih.gov/35112252","citation_count":6,"is_preprint":false},{"pmid":"38065968","id":"PMC_38065968","title":"FilGAP regulates tumor growth in Glioma through the regulation of mTORC1 and mTORC2.","date":"2023","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/38065968","citation_count":5,"is_preprint":false},{"pmid":"27130700","id":"PMC_27130700","title":"The RacGAP protein FilGAP is a negative regulator of chemokine-promoted lymphocyte migration.","date":"2016","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/27130700","citation_count":5,"is_preprint":false},{"pmid":"35433684","id":"PMC_35433684","title":"A Single-Nucleotide Polymorphism in the Promoter of Porcine ARHGAP24 Gene Regulates Aggressive Behavior of Weaned Pigs After Mixing by Affecting the Binding of Transcription Factor p53.","date":"2022","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/35433684","citation_count":4,"is_preprint":false},{"pmid":"31078260","id":"PMC_31078260","title":"FilGAP regulates distinct stages of epithelial tubulogenesis.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31078260","citation_count":3,"is_preprint":false},{"pmid":"21823009","id":"PMC_21823009","title":"Effects of variant rs346473 in ARHGAP24 gene on disease progression of HBV infection in Han Chinese population.","date":"2011","source":"Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban","url":"https://pubmed.ncbi.nlm.nih.gov/21823009","citation_count":3,"is_preprint":false},{"pmid":"38421271","id":"PMC_38421271","title":"FilGAP controls cell-extracellular matrix adhesion and process formation of kidney podocytes.","date":"2024","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/38421271","citation_count":2,"is_preprint":false},{"pmid":"38426123","id":"PMC_38426123","title":"RSK/GSK3-mediated phosphorylation of FilGAP regulates chemotactic cancer invasion.","date":"2024","source":"PNAS nexus","url":"https://pubmed.ncbi.nlm.nih.gov/38426123","citation_count":2,"is_preprint":false},{"pmid":"36641997","id":"PMC_36641997","title":"A t(4;13)(q21;q14) translocation in B-cell chronic lymphocytic leukemia causing concomitant homozygous DLEU2/miR15a/miR16-1 and heterozygous ARHGAP24 deletions.","date":"2023","source":"Cancer genetics","url":"https://pubmed.ncbi.nlm.nih.gov/36641997","citation_count":1,"is_preprint":false},{"pmid":"38074998","id":"PMC_38074998","title":"Knockdown of ARHGAP24 reduces intimal hyperplasia through inhibiting the proliferation and phenotypic switching of smooth muscle cells possibly by inactivating both AKT and ERK1/2 signaling pathways.","date":"2023","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/38074998","citation_count":1,"is_preprint":false},{"pmid":"37482421","id":"PMC_37482421","title":"FilGAP, a GAP for Rac1, down-regulates invadopodia formation in breast cancer cells.","date":"2023","source":"Cell structure and function","url":"https://pubmed.ncbi.nlm.nih.gov/37482421","citation_count":1,"is_preprint":false},{"pmid":"40498520","id":"PMC_40498520","title":"Novel polymorphism at ARHGAP24 gene and its association with growth traits in Hu sheep.","date":"2025","source":"Animal biotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/40498520","citation_count":0,"is_preprint":false},{"pmid":"40029073","id":"PMC_40029073","title":"Impairment of endocytosis-related factors FNBP1L, ARHGAP24, and ATP6V1B1 increases HIV-1 entry into dendritic cells.","date":"2025","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/40029073","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2024.10.16.24315096","title":"Isolated prolapse of the posterior mitral valve leaflet: phenotypic refinement, heritability and genetic etiology","date":"2024-10-17","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.16.24315096","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":24945,"output_tokens":6316,"usd":0.084788,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14928,"output_tokens":4876,"usd":0.09827,"stage2_stop_reason":"end_turn"},"total_usd":0.183058,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2006,\n      \"finding\": \"FilGAP (ARHGAP24) is a Rac-specific GTPase-activating protein (GAP) that binds filamin A (FLNa). FLNa targets FilGAP to sites of membrane protrusion where it antagonizes Rac in vivo. ROCK phosphorylates FilGAP, and this phosphorylation stimulates its RacGAP activity and is required for FilGAP-mediated bleb formation. Knockdown of FilGAP abrogates ROCK-dependent suppression of lamellae, establishing FilGAP as a mediator of RhoA-Rac antagonism.\",\n      \"method\": \"siRNA knockdown, dominant-negative constructs, co-immunoprecipitation, in vitro kinase assay (ROCK phosphorylation), cell spreading assay on fibronectin, live-cell imaging of blebs\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal methods (biochemical binding, kinase assay, siRNA KD, dominant-negative), foundational paper replicated by subsequent work\",\n      \"pmids\": [\"16862148\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"FilGAP interacts with the 23rd Ig repeat of filamin A (IgFLNa23) via the C-terminal 32 residues of FilGAP engaging the C and D beta-strands of IgFLNa23. Tight complex requires dimerization of both partners and a flexible hinge between FLNa repeats 23-24. FilGAP does not bind FLNa homologs FLNb or FLNc. Disease-associated FLNa mutations disrupt this binding interface, perturb cell spreading, and weaken elasticity of the FLNa/F-actin network under mechanical stress.\",\n      \"method\": \"NMR structural analysis, in silico modeling, mutagenesis of interface residues, cell spreading assay, F-actin network mechanics\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — NMR structure combined with mutagenesis and functional validation, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"19293932\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"FilGAP is targeted to sites of force transfer by FLNa (specifically repeat 23). Force-induced redistribution of FilGAP is essential for suppression of Rac activity and lamellae formation in cells treated with tensile forces applied through integrins. Depletion of FilGAP by siRNA, inhibition by dominant-negative mutation, or deletion of its FLNa-binding domain all result in dramatic force-induced increase of annexin-V-positive (apoptotic) cells, establishing FilGAP as a mechanoprotective anti-apoptotic effector downstream of FLNa.\",\n      \"method\": \"siRNA knockdown, dominant-negative constructs, integrin-mediated force application, annexin-V staining for apoptosis, fluorescence microscopy\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KD/KO with defined cellular phenotype plus dominant-negative and deletion constructs, single lab\",\n      \"pmids\": [\"19144823\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Mechanical strain in reconstituted actin/FLNa networks differentially regulates binding of FLNa partners: strain increases β-integrin binding to FLNa while causing FilGAP to dissociate from FLNa. Both externally imposed bulk shear and myosin-II-driven forces were shown to regulate this differential binding using fluorescence loss after photoconversion (FLAP).\",\n      \"method\": \"Reconstituted minimal system (actin filaments + FLNa + β-integrin tail + FilGAP), fluorescence loss after photoconversion (FLAP), bulk shear application, myosin-II inhibition\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with purified components, novel biophysical method, published in Nature with rigorous controls\",\n      \"pmids\": [\"21926999\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Arhgap24 is upregulated in differentiating kidney podocytes and functions as a RhoA-activated Rac1-GAP in these cells. Arhgap24 knockdown increases active Rac1 and Cdc42 levels and alters podocyte cell shape and membrane dynamics. A disease-associated missense mutation in ARHGAP24 impairs its Rac1-GAP activity and co-segregates with familial focal segmental glomerulosclerosis (FSGS).\",\n      \"method\": \"siRNA knockdown in cultured podocytes, GTPase activation assays (active Rac1/Cdc42 pulldown), Sanger sequencing of FSGS patients, in vitro GAP activity assay of mutant protein\",\n      \"journal\": \"The Journal of clinical investigation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — in vitro GAP activity assay of mutant, cellular KD with GTPase measurements, human genetic validation, multiple orthogonal approaches\",\n      \"pmids\": [\"21911940\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"FilGAP is a mediator of Rho/ROCK-dependent amoeboid movement of carcinoma cells. Depletion of FilGAP induces elongated mesenchymal morphology; forced expression induces round/amoeboid morphology requiring Rho/ROCK-dependent phosphorylation of FilGAP. Depletion of FilGAP impairs breast cancer cell invasion through ECM and reduces tumor cell extravasation in vivo.\",\n      \"method\": \"siRNA knockdown, forced expression, cell morphology analysis, Matrigel invasion assay, in vivo extravasation assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KD and OE with defined morphological and invasion phenotypes, in vivo validation, single lab\",\n      \"pmids\": [\"23097497\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"FilGAP binds to activated Arf6 through its pleckstrin homology (PH) domain and is recruited to plasma membranes. Activated Arf6 stimulates the RacGAP activity of FilGAP. Knockdown of endogenous Arf6 by siRNA suppresses FilGAP-mediated bleb formation. A FilGAP mutant lacking PIP3 binding (R39C) still binds activated Arf6 and induces bleb formation, indicating a PIP3-independent interaction with Arf6.\",\n      \"method\": \"Co-immunoprecipitation, PH domain binding assay, siRNA knockdown, GAP activity assay, bleb formation assay, PI3K inhibitors\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical binding assay, GAP activity measurement, siRNA KD with functional readout, single lab multiple methods\",\n      \"pmids\": [\"24526684\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Phosphorylation of FilGAP serine 402 by ROCK regulates its subcellular localization: non-phosphorylatable FilGAP (S402A) localizes to actin cytoskeleton, while phosphomimetic or ROCK-phosphorylated FilGAP is diffusely cytoplasmic. Dephosphorylation of Ser-402 accompanies cell spreading on fibronectin. Calyculin A (Ser/Thr phosphatase inhibitor) suppresses spreading of WT FilGAP cells but not S402A cells. Arf6 activation stimulates bleb formation by both non-phosphorylatable and phosphomimetic FilGAP mutants.\",\n      \"method\": \"Site-directed mutagenesis (S402A, ST/A, ST/D), Phos-tag gel electrophoresis, fluorescence microscopy of localization, calyculin A treatment, cell spreading assay on fibronectin\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis with functional localization and spreading readouts, Phos-tag for phosphorylation state, single lab\",\n      \"pmids\": [\"26359494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"FilGAP promotes formation of adherens junctions in MDCK epithelial cells downstream of Rho-ROCK signaling. Knockdown of FilGAP stimulates HGF-induced disassembly and cell scattering; forced expression induces accumulation of E-cadherin at adherens junctions. The Rac GAP domain of FilGAP is required for suppression of HGF-induced scattering. ROCK-induced accumulation of E-cadherin at adherens junctions requires FilGAP and its ROCK-dependent phosphorylation (non-phosphorylatable FilGAP mutant fails to rescue).\",\n      \"method\": \"siRNA knockdown, forced expression, E-cadherin surface staining, HGF-induced scattering assay, ROCK overexpression, non-phosphorylatable mutant rescue\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean siRNA KD and OE with defined phenotypes, phospho-mutant analysis, single lab multiple orthogonal approaches\",\n      \"pmids\": [\"25908853\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"RBM10 is a FilGAP-interacting protein. Src family tyrosine kinase Fyn induces translocation of RBM10 from nucleus to cell periphery in a kinase-activity-dependent manner, where RBM10 co-localizes with FilGAP. RBM10 is required for targeting FilGAP to the cell periphery during spreading, and association of RBM10 with FilGAP stimulates FilGAP's RacGAP activity as shown by suppression of spreading, ruffle formation, and FilGAP-dependent protrusive structures.\",\n      \"method\": \"Co-immunoprecipitation (FilGAP-RBM10), siRNA knockdown of RBM10, forced expression of Fyn (WT and kinase-dead), fluorescence co-localization, cell spreading assay, ruffle formation assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, siRNA KD with functional readout, kinase-dead mutant control, single lab\",\n      \"pmids\": [\"26751795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"ARHGAP24 is required for regulation of ARF6-dependent pseudopod formation in breast carcinoma cells. Knockdown of ARHGAP24 increases the lifespan and length of pseudopods induced by activated ARF6. ARHGAP24 requires its ARF6-binding site to achieve ARF6-dependent actin remodeling.\",\n      \"method\": \"siRNA knockdown of ARHGAP24, activated ARF6 expression, pseudopod measurement by time-lapse microscopy, binding-site mutant analysis\",\n      \"journal\": \"Anticancer research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, siRNA KD with morphological readout, limited mechanistic depth\",\n      \"pmids\": [\"28870903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"AGAP1 (an Arf GAP) binds to the C-terminus of FilGAP through AGAP1's N-terminal GLD domain. FilGAP co-localizes with AGAP1 at intracellular vesicles, and FilGAP targeting to these vesicles requires its interaction with AGAP1. Depletion of AGAP1 causes accumulation of endogenous FilGAP at paxillin-positive focal adhesions and actin structures, suppresses cell spreading on collagen (relieved by co-depletion of FilGAP), and promotes cell invasion that is blocked by FilGAP depletion.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown (AGAP1, FilGAP, double KD), fluorescence co-localization, cell spreading assay on collagen, Matrigel invasion assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain mapping, genetic epistasis via double KD with defined functional readouts, single lab\",\n      \"pmids\": [\"31785816\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"FilGAP regulates distinct stages of MDCK epithelial tubulogenesis downstream of Rho-ROCK signaling. Depletion of FilGAP increases HGF-induced basolateral extensions from cysts; forced expression decreases extensions. Phosphomimetic FilGAP (ST/D) blocks membrane extension formation even in the presence of ROCK inhibitor Y-27632, and promotes lumen formation while blocking cell scattering in tubules.\",\n      \"method\": \"siRNA knockdown, forced expression, 3D MDCK cyst culture in ECM, HGF stimulation, ROCK inhibitor Y-27632, phosphomimetic mutant (ST/D)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, KD/OE with morphological readout, limited mechanistic depth beyond Rho-ROCK axis\",\n      \"pmids\": [\"31078260\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"FilGAP controls front-rear polarity and tumor cell migration speed through extracellular matrix. FilGAP localizes to the cell front through its PH domain in a PIP3-dependent manner to inactivate Rac at that site. Small GTPase Arf6 (binding to the FilGAP PH domain) also regulates FilGAP-mediated cell polarity and migration.\",\n      \"method\": \"Forced expression and siRNA depletion in breast cancer cells, collagen matrix migration assay, PH domain mutant analysis, PIP3 depletion, Arf6 manipulation\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mutant analysis combined with migration assay and localization studies, single lab, multiple methods\",\n      \"pmids\": [\"33710706\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CBX3 represses ARHGAP24 expression by forming a complex with TRIM28, TRIM24, and RBBP4, resulting in increased active Rac1 and promotion of lung adenocarcinoma progression.\",\n      \"method\": \"Co-immunoprecipitation of CBX3/TRIM28/TRIM24/RBBP4 complex, ChIP, Rac1 activity assay, ARHGAP24 expression measurement by western blot/qPCR\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP of multiprotein complex with functional validation of Rac1 activity, single lab\",\n      \"pmids\": [\"34785774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ARHGAP24 acts as a GTPase-independent scaffold in hepatocellular carcinoma to suppress β-catenin signaling. It recruits the E3 ubiquitin ligase WWP1 to promote degradation of pyruvate kinase M2 (PKM2). The C-terminal fragment of ARHGAP24 (aa 329-430) binds WWP1 and aa 631-748 binds PKM2 (aa 388-531). A GAP-deficient ARHGAP24 mutant exerts the same inhibitory effects as wild-type, confirming GAP-independent scaffolding function.\",\n      \"method\": \"Liquid chromatography-tandem mass spectrometry, co-immunoprecipitation, ubiquitination assay, GAP activity assay (GAP-deficient mutant), luciferase reporter for β-catenin, domain mapping with deletion constructs\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mass spectrometry-based binding partner identification, co-IP with domain mapping, ubiquitination assay, GAP-dead mutant control, single lab\",\n      \"pmids\": [\"36168627\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FilGAP interacts with mTORC1 via Raptor and with mTORC2 via Rictor and Sin1. Depletion of FilGAP decreases phosphorylation of S6K (mTORC1 substrate) and AKT (mTORC2 substrate) in KINGS-1 glioma cells; conversely, overexpression of FilGAP increases these phosphorylations, indicating FilGAP activates mTORC1/2. FilGAP depletion reduces spheroid growth.\",\n      \"method\": \"Co-immunoprecipitation (FilGAP with Raptor/Rictor/Sin1), siRNA knockdown, forced expression, western blot for p-S6K and p-AKT, PI3K inhibitor treatment, spheroid growth assay\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP with mTOR complex components, KD and OE with biochemical readouts, single lab\",\n      \"pmids\": [\"38065968\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FilGAP localizes to invadopodia through its PH domain binding to phosphatidylinositol 3,4-bisphosphate [PI(3,4)P2] and negatively regulates invadopodia formation by inactivating Rac1. Depletion of FilGAP or expression of GAP-deficient FilGAP mutant increases invadopodia formation and ECM degradation; overexpression suppresses both. PI(3,4)P2 depletion reduces FilGAP invadopodia localization.\",\n      \"method\": \"siRNA knockdown, forced expression, GAP-deficient mutant, PH domain PI(3,4)P2-binding mutant, gelatin degradation assay, EGF stimulation, fluorescence localization\",\n      \"journal\": \"Cell structure and function\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mutant analysis with functional and localization readouts, GAP-dead control, lipid-binding specificity tested, single lab\",\n      \"pmids\": [\"37482421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FilGAP is phosphorylated at Ser625 by p90 ribosomal S6 kinase (RSK) downstream of EGF signaling, followed by phosphorylation at Ser621 by GSK3 in a sequential manner. This phosphorylation induces dissociation of FilGAP from actin filaments via a novel actin-localization domain. Phosphorylation inhibits FilGAP's lamellipodia-suppression activity. Non-phosphorylatable FilGAP mutant reduces cell migration speed and persistence toward EGF gradient, establishing this cascade as a regulator of chemotactic tumor cell migration.\",\n      \"method\": \"In vitro kinase assays (RSK, GSK3), site-directed mutagenesis (S625A, S621A, non-phosphorylatable mutant), actin co-sedimentation, EGF stimulation, chemotaxis assay, fluorescence localization\",\n      \"journal\": \"PNAS nexus\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay with site-specific mutagenesis, actin binding assay, functional chemotaxis readout, multiple orthogonal methods in single study\",\n      \"pmids\": [\"38426123\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FilGAP controls cell-ECM adhesion and process formation in podocytes by suppressing Rac1/PAK1 signaling. FilGAP localizes to focal adhesions (FAs); its depletion decreases FA formation, impairs attachment to ECM, increases Rac1 activity, and reduces formation of major processes and foot process-like projections. These deficits are rescued by inhibition of Rac1 or PAK1, placing PAK1 as a downstream effector of FilGAP/Rac1 in podocyte morphology.\",\n      \"method\": \"siRNA knockdown, forced expression, Rac1 activity pulldown assay, focal adhesion staining (paxillin/vinculin), podocyte process formation assay, Rac1 inhibitor, PAK1 inhibitor rescue\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis via inhibitor rescue, KD with multiple cellular readouts, single lab\",\n      \"pmids\": [\"38421271\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Loss-of-function variants in ARHGAP24 found in families with isolated posterior mitral valve prolapse (PostMVP) impair cellular adhesion and mechanotransduction capacities in vitro. Silencing of ARHGAP24 in zebrafish leads to atrioventricular regurgitation, establishing a functional role for FilGAP in cardiac valve development/maintenance.\",\n      \"method\": \"Zebrafish morpholino silencing (in vivo valve phenotype), in vitro functional assays of FilGAP variants (cell adhesion, mechanotransduction), whole genome sequencing of families\",\n      \"journal\": \"bioRxiv (preprint)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo zebrafish model with defined phenotype, in vitro functional validation of human variants, single study, preprint\",\n      \"pmids\": [\"bio_10.1101_2024.10.16.24315096\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"ARHGAP24/FilGAP is a Rac1-specific GTPase-activating protein that binds filamin A (via IgFLNa23) and is activated by ROCK-mediated phosphorylation (including at Ser402) downstream of RhoA, thereby mediating Rho-Rac antagonism to control actin remodeling, cell spreading, bleb formation, adherens junctions, focal adhesions, and cell polarity; mechanical strain on FLNa causes FilGAP to dissociate from FLNa (mechanotransduction); FilGAP is additionally regulated by Arf6 (binding through its PH domain to PIP3/PI(3,4)P2), by EGF-driven RSK/GSK3 phosphorylation at Ser625/621 (promoting actin dissociation and chemotactic migration), and by RBM10/Fyn-kinase signaling; it also acts as a GTPase-independent scaffold recruiting WWP1 to degrade PKM2 and suppress β-catenin signaling, and activates mTORC1/2 via interactions with Raptor, Rictor, and Sin1.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"ARHGAP24 (FilGAP) is a Rac1-specific GTPase-activating protein that integrates RhoA/ROCK signaling with actin remodeling to enforce Rho-Rac antagonism during cell spreading, motility, and tissue morphogenesis [#0, #4]. It is targeted to sites of membrane protrusion and force transfer by binding the 23rd Ig repeat of filamin A through its C-terminal residues, an interaction requiring dimerization of both partners; mechanical strain on the actin/FLNa network causes FilGAP to dissociate, coupling mechanotransduction to local Rac inactivation and mechanoprotection against force-induced apoptosis [#1, #2, #3]. ROCK phosphorylation, including at Ser402, stimulates its RacGAP activity and controls its partition between the actin cytoskeleton and cytoplasm, with dephosphorylation accompanying cell spreading on fibronectin [#0, #7]. FilGAP localization is further directed by phosphoinositides and Arf6: its PH domain binds PIP3/PI(3,4)P2 and activated Arf6 to recruit it to the plasma membrane, cell front, and invadopodia, where it locally inactivates Rac1 to govern bleb and pseudopod formation, front-rear polarity, and ECM invasion [#6, #13, #17]. An EGF-driven RSK-then-GSK3 cascade phosphorylates Ser625/Ser621 to release FilGAP from actin and promote chemotactic migration, while RBM10/Fyn and AGAP1 control its peripheral and vesicular targeting [#18, #9, #11]. Through these activities FilGAP shapes adherens junctions, focal adhesions, and epithelial tubulogenesis downstream of Rho-ROCK [#8, #19], and beyond its GAP function it acts as a scaffold that recruits WWP1 to degrade PKM2 and suppress \\u03b2-catenin signaling and that engages Raptor, Rictor, and Sin1 to activate mTORC1/2 [#15, #16]. Loss of ARHGAP24 function causes familial focal segmental glomerulosclerosis through defective podocyte Rac1/PAK1 control [#4, #19].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established FilGAP as the molecular link that allows RhoA/ROCK signaling to suppress Rac-driven lamellae, defining a dedicated effector of Rho-Rac antagonism rather than treating the two GTPases as independently regulated.\",\n      \"evidence\": \"siRNA knockdown, dominant-negative constructs, co-IP with filamin A, in vitro ROCK kinase assay, and bleb/spreading imaging\",\n      \"pmids\": [\"16862148\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the structural basis of the FLNa interaction\", \"Phosphosites mediating ROCK activation not yet mapped\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined the structural interface tethering FilGAP to filamin A and showed disease FLNa mutations disrupt it, explaining how FilGAP is positioned at force-bearing actin sites.\",\n      \"evidence\": \"NMR structure of the IgFLNa23/FilGAP C-terminus complex, interface mutagenesis, and F-actin network mechanics; complemented by force-application studies showing force-induced redistribution is required to suppress Rac and protect against apoptosis\",\n      \"pmids\": [\"19293932\", \"19144823\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism converting FLNa conformational change into GAP activation not detailed\", \"Apoptosis link single-lab\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstrated directly that mechanical strain acts as a switch dissociating FilGAP from filamin A, providing a biophysical mechanism for mechanotransduction at the actin network.\",\n      \"evidence\": \"Reconstituted actin/FLNa/\\u03b2-integrin/FilGAP system with FLAP under bulk shear and myosin-II-driven forces\",\n      \"pmids\": [\"21926999\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative force thresholds in cells not established\", \"Downstream consequence of dissociation in vivo inferred\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Connected ARHGAP24 to human disease and a physiological cell type, showing its Rac1-GAP activity maintains podocyte architecture and that a GAP-impairing mutation causes familial FSGS.\",\n      \"evidence\": \"Podocyte siRNA knockdown with active Rac1/Cdc42 pulldowns, Sanger sequencing of FSGS families, and in vitro GAP assay of mutant protein\",\n      \"pmids\": [\"21911940\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not connect podocyte function to downstream effector (later resolved as PAK1)\", \"RhoA-activation mechanism in podocytes not detailed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified Arf6 as an activator and membrane-recruitment partner acting through the FilGAP PH domain, separating PIP3 binding from Arf6 binding as distinct regulatory inputs.\",\n      \"evidence\": \"Co-IP, PH-domain binding assays, GAP activity measurement, siRNA Arf6 depletion, and PIP3-binding mutant (R39C) bleb assays\",\n      \"pmids\": [\"24526684\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of Arf6/PH interaction not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Showed ROCK phosphorylation of Ser402 acts as a localization switch between actin and cytoplasm, and that FilGAP promotes adherens junction assembly, extending its role to epithelial integrity.\",\n      \"evidence\": \"S402A/phosphomimetic mutants with Phos-tag and localization imaging; MDCK E-cadherin staining, HGF scattering, and phospho-mutant rescue\",\n      \"pmids\": [\"26359494\", \"25908853\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Phosphatase responsible for Ser402 dephosphorylation not identified\", \"Direct GAP target at junctions not measured\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Revealed RBM10 as a Fyn-regulated partner that relocates from nucleus to periphery to target FilGAP and stimulate its RacGAP activity, adding a tyrosine-kinase-controlled targeting layer.\",\n      \"evidence\": \"Reciprocal co-IP, RBM10 siRNA, WT/kinase-dead Fyn expression, and spreading/ruffle assays\",\n      \"pmids\": [\"26751795\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Fyn phosphorylates FilGAP directly not shown\", \"Physiological context of RBM10 relocation unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Placed FilGAP downstream of Arf6 in controlling pseudopod dynamics in carcinoma cells, linking its Arf6-binding site to actin remodeling during invasion.\",\n      \"evidence\": \"ARHGAP24 siRNA with activated Arf6 expression, time-lapse pseudopod measurement, and binding-site mutant\",\n      \"pmids\": [\"28870903\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Limited mechanistic depth, morphological readout only\", \"Single lab, low confidence\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified AGAP1 as a partner directing FilGAP to intracellular vesicles and away from focal adhesions, with epistasis defining FilGAP as the effector of AGAP1-controlled spreading and invasion.\",\n      \"evidence\": \"Co-IP with GLD-domain mapping, single and double siRNA knockdown, spreading on collagen, and Matrigel invasion; complemented by 3D tubulogenesis KD/OE studies\",\n      \"pmids\": [\"31785816\", \"31078260\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How vesicular vs FA targeting is balanced not quantified\", \"Tubulogenesis study low confidence\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined FilGAP as a determinant of front-rear polarity, recruited to the cell front by PIP3 and Arf6 to locally inactivate Rac and tune migration speed, and showed its expression is epigenetically repressed in cancer.\",\n      \"evidence\": \"PH-domain mutant migration assays with PIP3/Arf6 manipulation in breast cancer cells; CBX3/TRIM28/TRIM24/RBBP4 complex co-IP, ChIP, and Rac1 activity in lung adenocarcinoma\",\n      \"pmids\": [\"33710706\", \"34785774\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Spatial coordination of PIP3 and Arf6 inputs not resolved\", \"CBX3 repression mechanism single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Uncovered a GAP-independent scaffolding function in which FilGAP bridges WWP1 to PKM2 for degradation and suppresses \\u03b2-catenin signaling, expanding its role beyond Rac regulation.\",\n      \"evidence\": \"LC-MS/MS partner identification, co-IP with domain mapping, ubiquitination assay, \\u03b2-catenin luciferase reporter, and GAP-deficient mutant control\",\n      \"pmids\": [\"36168627\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"In vivo relevance to hepatocellular carcinoma not fully established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected FilGAP to growth signaling and invadopodia, showing it activates mTORC1/2 via Raptor/Rictor/Sin1 while its PI(3,4)P2-driven invadopodial localization restrains Rac1-dependent ECM degradation.\",\n      \"evidence\": \"Co-IP with mTOR components plus p-S6K/p-AKT and spheroid readouts; invadopodia gelatin degradation with PH-domain and GAP-dead mutants and PI(3,4)P2 depletion\",\n      \"pmids\": [\"38065968\", \"37482421\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether mTOR activation is GAP-dependent unresolved\", \"Direct biochemical link between FilGAP and mTOR activation unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Mapped a sequential RSK-then-GSK3 phosphorylation cascade at Ser625/Ser621 that releases FilGAP from actin to drive EGF chemotaxis, and tied FilGAP/Rac1/PAK1 to podocyte adhesion and cardiac valve integrity.\",\n      \"evidence\": \"In vitro RSK/GSK3 kinase assays with site mutants and actin co-sedimentation plus chemotaxis; podocyte KD with Rac1/PAK1 inhibitor rescue; zebrafish silencing and human PostMVP variant functional assays (preprint)\",\n      \"pmids\": [\"38426123\", \"38421271\", \"bio_10.1101_2024.10.16.24315096\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphatase reversing Ser625/621 not identified\", \"PostMVP finding is a preprint with single zebrafish model\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How FilGAP's diverse regulatory inputs (FLNa mechanics, ROCK/RSK/GSK3 phosphorylation, Arf6 and phosphoinositide targeting) are integrated in real time, and how its GAP-dependent and GAP-independent scaffolding functions are partitioned, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unified structural model coupling phosphorylation, lipid, and FLNa binding to GAP activation\", \"Switch between Rac-GAP and WWP1/PKM2 or mTOR scaffolding roles undefined\", \"In vivo physiological hierarchy of regulatory inputs untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 4, 6]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [6, 13, 17]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [15, 16]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 18]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 13]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [7, 18]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 4, 13]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 15, 20]},\n      {\"term_id\": \"R-HSA-1474244\", \"supporting_discovery_ids\": [5, 17, 19]}\n    ],\n    \"complexes\": [\"mTORC1\", \"mTORC2\"],\n    \"partners\": [\"FLNA\", \"RBM10\", \"AGAP1\", \"WWP1\", \"PKM2\", \"RPTOR\", \"RICTOR\", \"MAPKAP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}