{"gene":"FGD3","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2000,"finding":"Mouse Fgd3 encodes a RhoGEF protein that activates Cdc42, leading to filopodia formation in fibroblasts upon microinjection; the protein contains adjacent RhoGEF and pleckstrin homology (PH) domains, a second carboxy-terminal PH domain, and a FYVE domain.","method":"Microinjection of Fgd3 into fibroblasts with actin cytoskeleton readout; domain architecture analysis","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct microinjection experiment with defined cellular phenotype (filopodia formation) and domain characterization, single lab","pmids":["10721717"],"is_preprint":false},{"year":2008,"finding":"FGD3 functions as a GEF for Cdc42 (elevating GTP-bound Cdc42 levels) and induces broad sheet-like protrusions (rather than the finger-like protrusions induced by FGD1) in HeLa Tet-Off cells upon inducible expression.","method":"Inducible expression in HeLa Tet-Off cells; GTP-bound Cdc42 pulldown assay; morphological analysis","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — inducible expression with active Cdc42 measurement and defined morphological phenotype, single lab, two orthogonal methods","pmids":["18363964"],"is_preprint":false},{"year":2008,"finding":"FGD3 inhibits cell migration when inducibly expressed in HeLa Tet-Off cells, in contrast to FGD1 which stimulates migration, establishing that these highly homologous GEFs have opposing effects on motility.","method":"Inducible expression in HeLa Tet-Off cells; cell migration assay","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — defined loss/gain-of-function phenotype with clear directional result, single lab","pmids":["18363964"],"is_preprint":false},{"year":2008,"finding":"FGD3 protein is targeted for proteasomal degradation by the SCF(FWD1/β-TrCP) ubiquitin ligase upon phosphorylation of serine residues in its conserved DSGIDS motif, analogous to the destruction of FGD1.","method":"Phosphorylation site mutagenesis; co-immunoprecipitation with SCF(FWD1/β-TrCP) components; proteasome inhibitor rescue experiments","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis of degron motif plus co-IP and pharmacological rescue, single lab","pmids":["18363964"],"is_preprint":false},{"year":2015,"finding":"A non-synonymous variant of FGD3 identified in Japanese Black cattle produces a mutant protein with reduced GEF activity toward Cdc42, linking loss of FGD3 Cdc42-GEF activity to skeletal dysplasia and growth plate cartilage defects.","method":"Positional cloning; GEF activity assay of wild-type vs. mutant FGD3 protein; expression analysis in growth plate cartilage","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro GEF activity assay comparing wild-type and mutant protein, single study","pmids":["26306008"],"is_preprint":false},{"year":2022,"finding":"FGD3 binds with the transcription factor HSF4; FGD3 silencing promotes HSF4 nuclear translocation and increases p65 (RelA) expression, thereby activating canonical NF-κB signaling in pancreatic cancer cells.","method":"Co-immunoprecipitation (FGD3–HSF4 interaction); siRNA knockdown of FGD3 with immunoblotting and subcellular fractionation/immunofluorescence for HSF4 and p65","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus KD with two orthogonal readouts (nuclear translocation and protein level), single lab","pmids":["34975151"],"is_preprint":false},{"year":2025,"finding":"FGD3 mediates plasma membrane rupture (PMR) and lytic cell death (necrosis, necroptosis, pyroptosis) in breast cancer cells by controlling actin reorganization via the Cdc42–ARP2/3 axis; FGD3 also increases DAMP release and immunogenic calreticulin surface exposure, enhancing NK cell-mediated killing.","method":"Genome-wide CRISPR screen selecting against lytic cell death; 2D cell culture and patient-derived organoid experiments; orthotopic mouse xenografts; immunoblotting; immunofluorescence; NK-cell cytotoxicity assays","journal":"Journal of experimental & clinical cancer research : CR","confidence":"High","confidence_rationale":"Tier 2 / Strong — CRISPR screen followed by multiple orthogonal mechanistic validations (organoids, xenografts, cytotoxicity assays) identifying Cdc42-ARP2/3 as downstream effector","pmids":["41225536"],"is_preprint":false}],"current_model":"FGD3 is a Cdc42-specific guanine nucleotide exchange factor (GEF) that activates Cdc42 via conserved RhoGEF/PH domains to remodel the actin cytoskeleton—promoting filopodia or lamellipodia depending on cellular context and inhibiting cell migration—whose intracellular levels are controlled by SCF(FWD1/β-TrCP)-mediated proteasomal degradation through phosphorylation of a DSGIDS degron motif; additionally, FGD3 binds HSF4 to suppress NF-κB/p65 signaling, and at the plasma membrane it couples cell swelling to membrane rupture and lytic cell death via the Cdc42–ARP2/3 actin reorganization axis."},"narrative":{"mechanistic_narrative":"FGD3 is a Cdc42-specific guanine nucleotide exchange factor that remodels the actin cytoskeleton through its adjacent RhoGEF and pleckstrin homology domains [PMID:10721717, PMID:18363964]. By elevating GTP-bound Cdc42, it drives actin-based protrusions—filopodia in fibroblasts and broad sheet-like protrusions in HeLa cells—and, unlike its homolog FGD1, suppresses rather than promotes cell migration [PMID:10721717, PMID:18363964]. FGD3 protein abundance is restrained by SCF(FWD1/β-TrCP)-mediated proteasomal degradation following phosphorylation of serine residues in a conserved DSGIDS degron motif [PMID:18363964]. Loss of its Cdc42-GEF activity is linked to skeletal dysplasia and growth plate cartilage defects in cattle [PMID:26306008]. Beyond its cytoskeletal role, FGD3 binds the transcription factor HSF4 to restrain HSF4 nuclear translocation and canonical NF-κB (p65/RelA) signaling [PMID:34975151], and at the plasma membrane it couples cell swelling to membrane rupture and lytic cell death through a Cdc42–ARP2/3 actin reorganization axis, enhancing DAMP release and NK cell-mediated killing [PMID:41225536].","teleology":[{"year":2000,"claim":"Established that FGD3 is a RhoGEF acting on Cdc42, answering whether this protein could regulate the actin cytoskeleton.","evidence":"Microinjection of Fgd3 into fibroblasts with filopodia readout and domain architecture analysis","pmids":["10721717"],"confidence":"Medium","gaps":["Direct biochemical GEF activity not measured in this study","Endogenous function not addressed"]},{"year":2008,"claim":"Demonstrated FGD3 directly elevates active Cdc42 and produces a distinct protrusion morphology from FGD1, while opposing it in migration—showing homologous GEFs have divergent outputs.","evidence":"Inducible expression in HeLa Tet-Off cells with GTP-Cdc42 pulldown, morphological analysis, and migration assays","pmids":["18363964"],"confidence":"Medium","gaps":["Mechanism distinguishing FGD3 from FGD1 outputs not resolved","Migration effect studied only by overexpression"]},{"year":2008,"claim":"Identified how FGD3 levels are controlled, showing SCF(FWD1/β-TrCP) degrades phosphorylated FGD3 via a DSGIDS degron.","evidence":"Degron mutagenesis, co-IP with SCF components, and proteasome inhibitor rescue","pmids":["18363964"],"confidence":"Medium","gaps":["Upstream kinase phosphorylating the degron not identified","Physiological trigger for degradation unknown"]},{"year":2015,"claim":"Linked FGD3 Cdc42-GEF activity to organismal phenotype, showing a variant with reduced GEF activity causes skeletal dysplasia.","evidence":"Positional cloning in cattle, in vitro GEF activity comparison of wild-type vs mutant, and growth plate expression analysis","pmids":["26306008"],"confidence":"Medium","gaps":["Cell-type mechanism in chondrocytes not defined","Human disease relevance not established"]},{"year":2022,"claim":"Revealed a non-cytoskeletal role, showing FGD3 binds HSF4 to suppress NF-κB signaling in pancreatic cancer cells.","evidence":"Co-IP for FGD3–HSF4 binding and siRNA knockdown with fractionation/IF for HSF4 and p65","pmids":["34975151"],"confidence":"Medium","gaps":["Whether GEF activity is required for HSF4 binding unknown","Direct binding interface not mapped"]},{"year":2025,"claim":"Established FGD3 as a driver of plasma membrane rupture and immunogenic lytic cell death via the Cdc42–ARP2/3 axis.","evidence":"Genome-wide CRISPR screen, organoids, orthotopic xenografts, and NK-cell cytotoxicity assays","pmids":["41225536"],"confidence":"High","gaps":["Molecular link between swelling sensing and FGD3 activation unclear","How actin reorganization mechanically drives rupture not detailed"]},{"year":null,"claim":"How FGD3's distinct activities—cytoskeletal GEF function, HSF4/NF-κB regulation, and membrane rupture—are integrated and contextually selected remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of FGD3 domain function","Upstream signals selecting between filopodia, lamellipodia, and lytic death pathways unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,4]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,1,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6]}],"pathway":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[6]}],"complexes":[],"partners":["CDC42","HSF4","BTRC","ARPC"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q5JSP0","full_name":"FYVE, RhoGEF and PH domain-containing protein 3","aliases":["Zinc finger FYVE domain-containing protein 5"],"length_aa":725,"mass_kda":79.4,"function":"Promotes the formation of filopodia. May activate CDC42, a member of the Ras-like family of Rho- and Rac proteins, by exchanging bound GDP for free GTP. Plays a role in regulating the actin cytoskeleton and cell shape (By similarity)","subcellular_location":"Cytoplasm; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q5JSP0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/FGD3","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/FGD3","total_profiled":1310},"omim":[{"mim_id":"617554","title":"FYVE, RhoGEF, AND PH DOMAIN-CONTAINING PROTEIN 3; FGD3","url":"https://www.omim.org/entry/617554"},{"mim_id":"609197","title":"GLUCOCORTICOID DEFICIENCY 3; GCCD3","url":"https://www.omim.org/entry/609197"},{"mim_id":"300358","title":"PROTEIN KINASE, LYSINE-DEFICIENT 3; WNK3","url":"https://www.omim.org/entry/300358"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":37.3},{"tissue":"lymphoid tissue","ntpm":32.5}],"url":"https://www.proteinatlas.org/search/FGD3"},"hgnc":{"alias_symbol":["FLJ00004","ZFYVE5"],"prev_symbol":[]},"alphafold":{"accession":"Q5JSP0","domains":[{"cath_id":"1.20.900.10","chopping":"154-357","consensus_level":"high","plddt":87.7727,"start":154,"end":357},{"cath_id":"2.30.29.30","chopping":"370-480","consensus_level":"high","plddt":86.4206,"start":370,"end":480},{"cath_id":"-","chopping":"543-585","consensus_level":"medium","plddt":79.29,"start":543,"end":585},{"cath_id":"2.30.29.30","chopping":"605-702","consensus_level":"high","plddt":81.2163,"start":605,"end":702}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5JSP0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q5JSP0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q5JSP0-F1-predicted_aligned_error_v6.png","plddt_mean":68.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=FGD3","jax_strain_url":"https://www.jax.org/strain/search?query=FGD3"},"sequence":{"accession":"Q5JSP0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q5JSP0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q5JSP0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5JSP0"}},"corpus_meta":[{"pmid":"18363964","id":"PMC_18363964","title":"Novel insights into FGD3, a putative GEF for Cdc42, that undergoes SCF(FWD1/beta-TrCP)-mediated proteasomal degradation analogous to that of its homologue FGD1 but regulates cell morphology and motility differently from FGD1.","date":"2008","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/18363964","citation_count":46,"is_preprint":false},{"pmid":"10721717","id":"PMC_10721717","title":"Isolation, characterization, and mapping of the mouse Fgd3 gene, a new Faciogenital Dysplasia (FGD1; Aarskog Syndrome) gene homologue.","date":"2000","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/10721717","citation_count":35,"is_preprint":false},{"pmid":"26306008","id":"PMC_26306008","title":"Non-synonymous FGD3 Variant as Positional Candidate for Disproportional Tall Stature Accounting for a Carcass Weight QTL (CW-3) and Skeletal Dysplasia in Japanese Black Cattle.","date":"2015","source":"PLoS genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26306008","citation_count":26,"is_preprint":false},{"pmid":"34975151","id":"PMC_34975151","title":"FGD3 binds with HSF4 to suppress p65 expression and inhibit pancreatic cancer progression.","date":"2022","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/34975151","citation_count":13,"is_preprint":false},{"pmid":"41225536","id":"PMC_41225536","title":"FGD3 mediates lytic cell death, enhancing efficacy and immunogenicity of chemotherapy agents in breast cancer.","date":"2025","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/41225536","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5010,"output_tokens":1784,"usd":0.020895,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8634,"output_tokens":2139,"usd":0.048322,"stage2_stop_reason":"end_turn"},"total_usd":0.069217,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"Mouse Fgd3 encodes a RhoGEF protein that activates Cdc42, leading to filopodia formation in fibroblasts upon microinjection; the protein contains adjacent RhoGEF and pleckstrin homology (PH) domains, a second carboxy-terminal PH domain, and a FYVE domain.\",\n      \"method\": \"Microinjection of Fgd3 into fibroblasts with actin cytoskeleton readout; domain architecture analysis\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct microinjection experiment with defined cellular phenotype (filopodia formation) and domain characterization, single lab\",\n      \"pmids\": [\"10721717\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"FGD3 functions as a GEF for Cdc42 (elevating GTP-bound Cdc42 levels) and induces broad sheet-like protrusions (rather than the finger-like protrusions induced by FGD1) in HeLa Tet-Off cells upon inducible expression.\",\n      \"method\": \"Inducible expression in HeLa Tet-Off cells; GTP-bound Cdc42 pulldown assay; morphological analysis\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — inducible expression with active Cdc42 measurement and defined morphological phenotype, single lab, two orthogonal methods\",\n      \"pmids\": [\"18363964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"FGD3 inhibits cell migration when inducibly expressed in HeLa Tet-Off cells, in contrast to FGD1 which stimulates migration, establishing that these highly homologous GEFs have opposing effects on motility.\",\n      \"method\": \"Inducible expression in HeLa Tet-Off cells; cell migration assay\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — defined loss/gain-of-function phenotype with clear directional result, single lab\",\n      \"pmids\": [\"18363964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"FGD3 protein is targeted for proteasomal degradation by the SCF(FWD1/β-TrCP) ubiquitin ligase upon phosphorylation of serine residues in its conserved DSGIDS motif, analogous to the destruction of FGD1.\",\n      \"method\": \"Phosphorylation site mutagenesis; co-immunoprecipitation with SCF(FWD1/β-TrCP) components; proteasome inhibitor rescue experiments\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis of degron motif plus co-IP and pharmacological rescue, single lab\",\n      \"pmids\": [\"18363964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"A non-synonymous variant of FGD3 identified in Japanese Black cattle produces a mutant protein with reduced GEF activity toward Cdc42, linking loss of FGD3 Cdc42-GEF activity to skeletal dysplasia and growth plate cartilage defects.\",\n      \"method\": \"Positional cloning; GEF activity assay of wild-type vs. mutant FGD3 protein; expression analysis in growth plate cartilage\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro GEF activity assay comparing wild-type and mutant protein, single study\",\n      \"pmids\": [\"26306008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"FGD3 binds with the transcription factor HSF4; FGD3 silencing promotes HSF4 nuclear translocation and increases p65 (RelA) expression, thereby activating canonical NF-κB signaling in pancreatic cancer cells.\",\n      \"method\": \"Co-immunoprecipitation (FGD3–HSF4 interaction); siRNA knockdown of FGD3 with immunoblotting and subcellular fractionation/immunofluorescence for HSF4 and p65\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus KD with two orthogonal readouts (nuclear translocation and protein level), single lab\",\n      \"pmids\": [\"34975151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FGD3 mediates plasma membrane rupture (PMR) and lytic cell death (necrosis, necroptosis, pyroptosis) in breast cancer cells by controlling actin reorganization via the Cdc42–ARP2/3 axis; FGD3 also increases DAMP release and immunogenic calreticulin surface exposure, enhancing NK cell-mediated killing.\",\n      \"method\": \"Genome-wide CRISPR screen selecting against lytic cell death; 2D cell culture and patient-derived organoid experiments; orthotopic mouse xenografts; immunoblotting; immunofluorescence; NK-cell cytotoxicity assays\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CRISPR screen followed by multiple orthogonal mechanistic validations (organoids, xenografts, cytotoxicity assays) identifying Cdc42-ARP2/3 as downstream effector\",\n      \"pmids\": [\"41225536\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"FGD3 is a Cdc42-specific guanine nucleotide exchange factor (GEF) that activates Cdc42 via conserved RhoGEF/PH domains to remodel the actin cytoskeleton—promoting filopodia or lamellipodia depending on cellular context and inhibiting cell migration—whose intracellular levels are controlled by SCF(FWD1/β-TrCP)-mediated proteasomal degradation through phosphorylation of a DSGIDS degron motif; additionally, FGD3 binds HSF4 to suppress NF-κB/p65 signaling, and at the plasma membrane it couples cell swelling to membrane rupture and lytic cell death via the Cdc42–ARP2/3 actin reorganization axis.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"FGD3 is a Cdc42-specific guanine nucleotide exchange factor that remodels the actin cytoskeleton through its adjacent RhoGEF and pleckstrin homology domains [#0, #1]. By elevating GTP-bound Cdc42, it drives actin-based protrusions—filopodia in fibroblasts and broad sheet-like protrusions in HeLa cells—and, unlike its homolog FGD1, suppresses rather than promotes cell migration [#0, #1, #2]. FGD3 protein abundance is restrained by SCF(FWD1/β-TrCP)-mediated proteasomal degradation following phosphorylation of serine residues in a conserved DSGIDS degron motif [#3]. Loss of its Cdc42-GEF activity is linked to skeletal dysplasia and growth plate cartilage defects in cattle [#4]. Beyond its cytoskeletal role, FGD3 binds the transcription factor HSF4 to restrain HSF4 nuclear translocation and canonical NF-κB (p65/RelA) signaling [#5], and at the plasma membrane it couples cell swelling to membrane rupture and lytic cell death through a Cdc42–ARP2/3 actin reorganization axis, enhancing DAMP release and NK cell-mediated killing [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established that FGD3 is a RhoGEF acting on Cdc42, answering whether this protein could regulate the actin cytoskeleton.\",\n      \"evidence\": \"Microinjection of Fgd3 into fibroblasts with filopodia readout and domain architecture analysis\",\n      \"pmids\": [\"10721717\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical GEF activity not measured in this study\", \"Endogenous function not addressed\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Demonstrated FGD3 directly elevates active Cdc42 and produces a distinct protrusion morphology from FGD1, while opposing it in migration—showing homologous GEFs have divergent outputs.\",\n      \"evidence\": \"Inducible expression in HeLa Tet-Off cells with GTP-Cdc42 pulldown, morphological analysis, and migration assays\",\n      \"pmids\": [\"18363964\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism distinguishing FGD3 from FGD1 outputs not resolved\", \"Migration effect studied only by overexpression\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified how FGD3 levels are controlled, showing SCF(FWD1/β-TrCP) degrades phosphorylated FGD3 via a DSGIDS degron.\",\n      \"evidence\": \"Degron mutagenesis, co-IP with SCF components, and proteasome inhibitor rescue\",\n      \"pmids\": [\"18363964\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Upstream kinase phosphorylating the degron not identified\", \"Physiological trigger for degradation unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Linked FGD3 Cdc42-GEF activity to organismal phenotype, showing a variant with reduced GEF activity causes skeletal dysplasia.\",\n      \"evidence\": \"Positional cloning in cattle, in vitro GEF activity comparison of wild-type vs mutant, and growth plate expression analysis\",\n      \"pmids\": [\"26306008\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Cell-type mechanism in chondrocytes not defined\", \"Human disease relevance not established\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Revealed a non-cytoskeletal role, showing FGD3 binds HSF4 to suppress NF-κB signaling in pancreatic cancer cells.\",\n      \"evidence\": \"Co-IP for FGD3–HSF4 binding and siRNA knockdown with fractionation/IF for HSF4 and p65\",\n      \"pmids\": [\"34975151\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether GEF activity is required for HSF4 binding unknown\", \"Direct binding interface not mapped\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established FGD3 as a driver of plasma membrane rupture and immunogenic lytic cell death via the Cdc42–ARP2/3 axis.\",\n      \"evidence\": \"Genome-wide CRISPR screen, organoids, orthotopic xenografts, and NK-cell cytotoxicity assays\",\n      \"pmids\": [\"41225536\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between swelling sensing and FGD3 activation unclear\", \"How actin reorganization mechanically drives rupture not detailed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How FGD3's distinct activities—cytoskeletal GEF function, HSF4/NF-κB regulation, and membrane rupture—are integrated and contextually selected remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of FGD3 domain function\", \"Upstream signals selecting between filopodia, lamellipodia, and lytic death pathways unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 4]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 1, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"CDC42\", \"HSF4\", \"BTRC\", \"ARPC\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}