{"gene":"DENND1C","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2010,"finding":"DENND1C (along with DENND1A and DENND1B) was identified as a Rab GEF (guanine nucleotide exchange factor) that specifically activates Rab35, placing it in the family of DENN-domain GEFs responsible for site-specific Rab activation in membrane trafficking.","method":"Systematic family-wide characterization of 17 human DENN domain proteins including GEF activity assays and localization studies","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — systematic biochemical characterization across entire protein family, replicated findings for multiple DENN members including DENND1C as Rab35 GEF","pmids":["20937701"],"is_preprint":false},{"year":2011,"finding":"DENND1C (connecdenn 3) directly binds actin via a unique C-terminal motif not found in DENND1A or DENND1B, coupling Rab35 activation to the actin cytoskeleton and causing formation of protrusive membrane extensions. Both the actin-binding C-terminal motif and the N-terminal DENN domain (GEF activity) are required for this effect. DENND1C and active Rab35 colocalize with the actin-bundling protein fascin on actin filaments, identifying DENND1C as the relevant GEF for Rab35-fascin-mediated actin regulation.","method":"Direct actin-binding assays, domain deletion/mutagenesis, co-localization imaging, cell shape phenotype readout (protrusive membrane extensions)","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro direct binding assays, mutagenesis of specific motif, multiple orthogonal methods in single focused study on DENND1C","pmids":["22072793"],"is_preprint":false},{"year":2014,"finding":"In T lymphocytes stimulated with chemokine, the kinase Mst1 phosphorylates DENND1C, which activates Rab13. Active Rab13 then associates with Mst1 to facilitate delivery of the integrin LFA-1 to the leading edge of migrating lymphocytes via myosin Va along actin filaments, promoting lymphocyte adhesion and trafficking.","method":"Phosphorylation assay (Mst1-dependent phosphorylation of DENND1C), Rab13 activation assay, co-immunoprecipitation, cell migration/adhesion assay on ICAM-1, Rab13-deficient mouse model","journal":"Science signaling","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, phosphorylation assay, in vivo mouse KO with defined cellular phenotype, multiple orthogonal methods in single study","pmids":["25074980"],"is_preprint":false},{"year":2019,"finding":"Upon IL-17A stimulation of airway smooth muscle cells, DENND1C (as GEF) is recruited together with Rab35 to the IL-17R/Act1 receptor complex, leading to Rab35 activation, which is required for PKCα activation and fascin phosphorylation at Ser39, enabling stress fiber formation and enhanced smooth muscle contraction.","method":"Co-immunoprecipitation of DENND1C/Rab35 with IL-17R/Act1 complex, Rab35 knockdown with PKCα activation readout, fascin phosphorylation assay, airway smooth muscle contraction assay","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP recruitment assay, siRNA knockdown with defined downstream phosphorylation and contractility readouts, single lab","pmids":["30683702"],"is_preprint":false},{"year":2020,"finding":"Among the four known Rab35 GEFs (DENND1A, DENND1B, DENND1C, folliculin), DENND1C knockout did not phenocopy Rab35 knockout in podocalyxin (PODXL) trafficking in either 2D or 3D MDCK epithelial cell cultures, indicating that DENND1C is not the primary GEF for Rab35 in polarized epithelial PODXL trafficking (negative finding for DENND1C in this context).","method":"DENND1C knockout in MDCK II cells, fluorescence imaging of podocalyxin localization in 2D and 3D cultures","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KO with defined trafficking readout, single lab, negative finding specifically for DENND1C in this pathway","pmids":["31992598"],"is_preprint":false}],"current_model":"DENND1C (connecdenn 3) is a DENN-domain guanine nucleotide exchange factor (GEF) that activates both Rab35 and Rab13: it activates Rab35 via its N-terminal DENN domain while uniquely binding actin through a C-terminal motif to couple Rab35 activation to cytoskeletal remodeling and protrusive membrane formation; it is phosphorylated by the kinase Mst1 to activate Rab13 for LFA-1 integrin delivery to the leading edge of migrating lymphocytes; and it is recruited to the IL-17R/Act1 complex to activate Rab35-PKCα signaling in airway smooth muscle contraction."},"narrative":{"mechanistic_narrative":"DENND1C (connecdenn 3) is a DENN-domain guanine nucleotide exchange factor that couples site-specific Rab GTPase activation to actin-based membrane remodeling and cell motility [PMID:20937701, PMID:22072793]. It activates Rab35 through its N-terminal DENN domain, and unlike its paralogs DENND1A and DENND1B it possesses a unique C-terminal actin-binding motif; both the DENN domain and this motif are required to drive formation of protrusive membrane extensions, where DENND1C and active Rab35 colocalize with the actin-bundling protein fascin [PMID:22072793]. In a parallel signaling context, DENND1C is phosphorylated by the kinase Mst1 to activate Rab13, which then promotes myosin Va-dependent delivery of the integrin LFA-1 to the leading edge of chemokine-stimulated migrating lymphocytes [PMID:25074980]. DENND1C is also recruited with Rab35 to the IL-17R/Act1 complex upon IL-17A stimulation, driving Rab35-dependent PKCα activation and fascin Ser39 phosphorylation to enable stress fiber formation and airway smooth muscle contraction [PMID:30683702]. Its GEF activity is context-restricted: DENND1C knockout does not phenocopy Rab35 loss in polarized epithelial podocalyxin trafficking, indicating it is not the primary Rab35 GEF in that pathway [PMID:31992598].","teleology":[{"year":2010,"claim":"Establishing DENND1C as a Rab GEF answered which Rab it acts on, placing it within the DENN-domain family responsible for site-specific Rab activation in membrane trafficking.","evidence":"Family-wide GEF activity assays and localization across 17 human DENN domain proteins","pmids":["20937701"],"confidence":"High","gaps":["Does not define a cellular pathway or membrane compartment specific to DENND1C","Does not distinguish DENND1C function from DENND1A/DENND1B"]},{"year":2011,"claim":"Identifying a unique C-terminal actin-binding motif explained how DENND1C distinctively couples Rab35 activation to cytoskeletal remodeling and protrusive membrane formation, separating it functionally from its paralogs.","evidence":"Direct actin-binding assays, domain deletion/mutagenesis, colocalization imaging, and cell-shape phenotype in a focused DENND1C study","pmids":["22072793"],"confidence":"High","gaps":["Physiological context of protrusive extension formation not defined","Mechanism linking fascin to the Rab35-DENND1C complex not resolved"]},{"year":2014,"claim":"Discovery of Mst1-dependent phosphorylation activating Rab13 revealed a second GTPase target and a kinase-regulated role for DENND1C in integrin delivery during lymphocyte migration.","evidence":"Phosphorylation and Rab13 activation assays, reciprocal co-IP, migration/adhesion assays, and Rab13-deficient mouse model","pmids":["25074980"],"confidence":"High","gaps":["Phosphorylation site(s) on DENND1C not mapped","How phosphorylation switches GEF specificity toward Rab13 unresolved"]},{"year":2019,"claim":"Placing DENND1C/Rab35 downstream of the IL-17R/Act1 complex connected its GEF activity to receptor signaling driving PKCα-fascin-dependent smooth muscle contraction.","evidence":"Co-IP of recruitment to IL-17R/Act1, Rab35 knockdown with PKCα and fascin phosphorylation readouts, and contraction assays in airway smooth muscle cells","pmids":["30683702"],"confidence":"Medium","gaps":["Single-lab finding without independent replication","Direct versus indirect recruitment of DENND1C to the complex not distinguished"]},{"year":2020,"claim":"Testing DENND1C against other Rab35 GEFs established that its activity is context-restricted, ruling it out as the primary GEF for epithelial podocalyxin trafficking.","evidence":"DENND1C knockout in MDCK II cells with podocalyxin localization imaging in 2D and 3D","pmids":["31992598"],"confidence":"Medium","gaps":["Possible redundancy with other GEFs not excluded","Does not identify which contexts depend specifically on DENND1C"]},{"year":null,"claim":"How DENND1C selects between its Rab35 and Rab13 targets, and what determines its tissue- and context-specific deployment, remains unresolved.","evidence":"No single study reconciles the GEF target switch with upstream regulation","pmids":[],"confidence":"Medium","gaps":["Structural basis of dual Rab35/Rab13 specificity unknown","Phosphorylation sites and their effect on substrate choice unmapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[1]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,2]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,3]}],"complexes":[],"partners":["RAB35","RAB13","STK4","FASCIN"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IV53","full_name":"DENN domain-containing protein 1C","aliases":["Connecdenn 3","Protein FAM31C"],"length_aa":801,"mass_kda":87.1,"function":"Guanine nucleotide exchange factor (GEF) which may activate RAB8A, RAB13 and RAB35. Promotes the exchange of GDP to GTP, converting inactive GDP-bound Rab proteins into their active GTP-bound form","subcellular_location":"Cytoplasm, cytosol; Cytoplasmic vesicle, clathrin-coated vesicle","url":"https://www.uniprot.org/uniprotkb/Q8IV53/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/DENND1C","classification":"Not Classified","n_dependent_lines":16,"n_total_lines":1208,"dependency_fraction":0.013245033112582781},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/DENND1C","total_profiled":1310},"omim":[{"mim_id":"613634","title":"DENN/MADD DOMAIN-CONTAINING PROTEIN 1C; DENND1C","url":"https://www.omim.org/entry/613634"},{"mim_id":"613633","title":"DENN/MADD DOMAIN-CONTAINING PROTEIN 1A; DENND1A","url":"https://www.omim.org/entry/613633"},{"mim_id":"613292","title":"DENN/MADD DOMAIN-CONTAINING PROTEIN 1B; DENND1B","url":"https://www.omim.org/entry/613292"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"},{"location":"Nucleoplasm","reliability":"Additional"},{"location":"Centrosome","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":20.0},{"tissue":"intestine","ntpm":13.8},{"tissue":"lymphoid tissue","ntpm":34.2}],"url":"https://www.proteinatlas.org/search/DENND1C"},"hgnc":{"alias_symbol":["FLJ22757"],"prev_symbol":["FAM31C"]},"alphafold":{"accession":"Q8IV53","domains":[{"cath_id":"3.30.450.200","chopping":"16-138","consensus_level":"high","plddt":91.9631,"start":16,"end":138},{"cath_id":"3.40.50.11500","chopping":"191-335","consensus_level":"medium","plddt":95.9257,"start":191,"end":335},{"cath_id":"-","chopping":"341-430","consensus_level":"medium","plddt":85.2143,"start":341,"end":430}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IV53","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IV53-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IV53-F1-predicted_aligned_error_v6.png","plddt_mean":62.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=DENND1C","jax_strain_url":"https://www.jax.org/strain/search?query=DENND1C"},"sequence":{"accession":"Q8IV53","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IV53.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IV53/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IV53"}},"corpus_meta":[{"pmid":"20937701","id":"PMC_20937701","title":"Family-wide characterization of the DENN domain Rab GDP-GTP exchange factors.","date":"2010","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/20937701","citation_count":280,"is_preprint":false},{"pmid":"25651499","id":"PMC_25651499","title":"An analysis of DNA methylation in human adipose tissue reveals differential modification of obesity genes before and after gastric bypass and weight loss.","date":"2015","source":"Genome biology","url":"https://pubmed.ncbi.nlm.nih.gov/25651499","citation_count":200,"is_preprint":false},{"pmid":"25074980","id":"PMC_25074980","title":"Rab13 acts downstream of the kinase Mst1 to deliver the integrin LFA-1 to the cell surface for lymphocyte trafficking.","date":"2014","source":"Science signaling","url":"https://pubmed.ncbi.nlm.nih.gov/25074980","citation_count":61,"is_preprint":false},{"pmid":"22072793","id":"PMC_22072793","title":"Connecdenn 3/DENND1C binds actin linking Rab35 activation to the actin cytoskeleton.","date":"2011","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/22072793","citation_count":47,"is_preprint":false},{"pmid":"36588164","id":"PMC_36588164","title":"PCSK9 facilitates melanoma pathogenesis via a network regulating tumor immunity.","date":"2023","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/36588164","citation_count":34,"is_preprint":false},{"pmid":"27312686","id":"PMC_27312686","title":"Biological significance of genome-wide DNA methylation profiles in keloids.","date":"2016","source":"The Laryngoscope","url":"https://pubmed.ncbi.nlm.nih.gov/27312686","citation_count":24,"is_preprint":false},{"pmid":"30683702","id":"PMC_30683702","title":"IL-17A Recruits Rab35 to IL-17R to Mediate PKCα-Dependent Stress Fiber Formation and Airway Smooth Muscle Contractility.","date":"2019","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/30683702","citation_count":15,"is_preprint":false},{"pmid":"31992598","id":"PMC_31992598","title":"Rab35-GEFs, DENND1A and folliculin differentially regulate podocalyxin trafficking in two- and three-dimensional epithelial cell cultures.","date":"2020","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/31992598","citation_count":9,"is_preprint":false},{"pmid":"40306147","id":"PMC_40306147","title":"Identification of shared pathophysiological molecules of major psychiatric disorders: A comprehensive analysis of serum immune complex antigens before and after electroconvulsive therapy.","date":"2025","source":"Journal of neuroimmunology","url":"https://pubmed.ncbi.nlm.nih.gov/40306147","citation_count":1,"is_preprint":false},{"pmid":"41031217","id":"PMC_41031217","title":"Identification of Progression-Associated Biomarkers in Lung Cancer Based on the Integrated Analysis of RNA Sequencing Data From Platelets and Tumor Tissues.","date":"2025","source":"International journal of genomics","url":"https://pubmed.ncbi.nlm.nih.gov/41031217","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.10.11.25337587","title":"Machine Learning for Predicting and Maximizing the Response of Breast Cancer Patients to Neoadjuvant Therapy","date":"2025-10-14","source":"bioRxiv","url":"https://doi.org/10.1101/2025.10.11.25337587","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":8380,"output_tokens":1551,"usd":0.024202,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8210,"output_tokens":2110,"usd":0.0469,"stage2_stop_reason":"end_turn"},"total_usd":0.071102,"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\": 2010,\n      \"finding\": \"DENND1C (along with DENND1A and DENND1B) was identified as a Rab GEF (guanine nucleotide exchange factor) that specifically activates Rab35, placing it in the family of DENN-domain GEFs responsible for site-specific Rab activation in membrane trafficking.\",\n      \"method\": \"Systematic family-wide characterization of 17 human DENN domain proteins including GEF activity assays and localization studies\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — systematic biochemical characterization across entire protein family, replicated findings for multiple DENN members including DENND1C as Rab35 GEF\",\n      \"pmids\": [\"20937701\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"DENND1C (connecdenn 3) directly binds actin via a unique C-terminal motif not found in DENND1A or DENND1B, coupling Rab35 activation to the actin cytoskeleton and causing formation of protrusive membrane extensions. Both the actin-binding C-terminal motif and the N-terminal DENN domain (GEF activity) are required for this effect. DENND1C and active Rab35 colocalize with the actin-bundling protein fascin on actin filaments, identifying DENND1C as the relevant GEF for Rab35-fascin-mediated actin regulation.\",\n      \"method\": \"Direct actin-binding assays, domain deletion/mutagenesis, co-localization imaging, cell shape phenotype readout (protrusive membrane extensions)\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro direct binding assays, mutagenesis of specific motif, multiple orthogonal methods in single focused study on DENND1C\",\n      \"pmids\": [\"22072793\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In T lymphocytes stimulated with chemokine, the kinase Mst1 phosphorylates DENND1C, which activates Rab13. Active Rab13 then associates with Mst1 to facilitate delivery of the integrin LFA-1 to the leading edge of migrating lymphocytes via myosin Va along actin filaments, promoting lymphocyte adhesion and trafficking.\",\n      \"method\": \"Phosphorylation assay (Mst1-dependent phosphorylation of DENND1C), Rab13 activation assay, co-immunoprecipitation, cell migration/adhesion assay on ICAM-1, Rab13-deficient mouse model\",\n      \"journal\": \"Science signaling\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, phosphorylation assay, in vivo mouse KO with defined cellular phenotype, multiple orthogonal methods in single study\",\n      \"pmids\": [\"25074980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Upon IL-17A stimulation of airway smooth muscle cells, DENND1C (as GEF) is recruited together with Rab35 to the IL-17R/Act1 receptor complex, leading to Rab35 activation, which is required for PKCα activation and fascin phosphorylation at Ser39, enabling stress fiber formation and enhanced smooth muscle contraction.\",\n      \"method\": \"Co-immunoprecipitation of DENND1C/Rab35 with IL-17R/Act1 complex, Rab35 knockdown with PKCα activation readout, fascin phosphorylation assay, airway smooth muscle contraction assay\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP recruitment assay, siRNA knockdown with defined downstream phosphorylation and contractility readouts, single lab\",\n      \"pmids\": [\"30683702\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Among the four known Rab35 GEFs (DENND1A, DENND1B, DENND1C, folliculin), DENND1C knockout did not phenocopy Rab35 knockout in podocalyxin (PODXL) trafficking in either 2D or 3D MDCK epithelial cell cultures, indicating that DENND1C is not the primary GEF for Rab35 in polarized epithelial PODXL trafficking (negative finding for DENND1C in this context).\",\n      \"method\": \"DENND1C knockout in MDCK II cells, fluorescence imaging of podocalyxin localization in 2D and 3D cultures\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KO with defined trafficking readout, single lab, negative finding specifically for DENND1C in this pathway\",\n      \"pmids\": [\"31992598\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"DENND1C (connecdenn 3) is a DENN-domain guanine nucleotide exchange factor (GEF) that activates both Rab35 and Rab13: it activates Rab35 via its N-terminal DENN domain while uniquely binding actin through a C-terminal motif to couple Rab35 activation to cytoskeletal remodeling and protrusive membrane formation; it is phosphorylated by the kinase Mst1 to activate Rab13 for LFA-1 integrin delivery to the leading edge of migrating lymphocytes; and it is recruited to the IL-17R/Act1 complex to activate Rab35-PKCα signaling in airway smooth muscle contraction.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"DENND1C (connecdenn 3) is a DENN-domain guanine nucleotide exchange factor that couples site-specific Rab GTPase activation to actin-based membrane remodeling and cell motility [#0, #1]. It activates Rab35 through its N-terminal DENN domain, and unlike its paralogs DENND1A and DENND1B it possesses a unique C-terminal actin-binding motif; both the DENN domain and this motif are required to drive formation of protrusive membrane extensions, where DENND1C and active Rab35 colocalize with the actin-bundling protein fascin [#1]. In a parallel signaling context, DENND1C is phosphorylated by the kinase Mst1 to activate Rab13, which then promotes myosin Va-dependent delivery of the integrin LFA-1 to the leading edge of chemokine-stimulated migrating lymphocytes [#2]. DENND1C is also recruited with Rab35 to the IL-17R/Act1 complex upon IL-17A stimulation, driving Rab35-dependent PKCα activation and fascin Ser39 phosphorylation to enable stress fiber formation and airway smooth muscle contraction [#3]. Its GEF activity is context-restricted: DENND1C knockout does not phenocopy Rab35 loss in polarized epithelial podocalyxin trafficking, indicating it is not the primary Rab35 GEF in that pathway [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Establishing DENND1C as a Rab GEF answered which Rab it acts on, placing it within the DENN-domain family responsible for site-specific Rab activation in membrane trafficking.\",\n      \"evidence\": \"Family-wide GEF activity assays and localization across 17 human DENN domain proteins\",\n      \"pmids\": [\"20937701\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not define a cellular pathway or membrane compartment specific to DENND1C\", \"Does not distinguish DENND1C function from DENND1A/DENND1B\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Identifying a unique C-terminal actin-binding motif explained how DENND1C distinctively couples Rab35 activation to cytoskeletal remodeling and protrusive membrane formation, separating it functionally from its paralogs.\",\n      \"evidence\": \"Direct actin-binding assays, domain deletion/mutagenesis, colocalization imaging, and cell-shape phenotype in a focused DENND1C study\",\n      \"pmids\": [\"22072793\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological context of protrusive extension formation not defined\", \"Mechanism linking fascin to the Rab35-DENND1C complex not resolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Discovery of Mst1-dependent phosphorylation activating Rab13 revealed a second GTPase target and a kinase-regulated role for DENND1C in integrin delivery during lymphocyte migration.\",\n      \"evidence\": \"Phosphorylation and Rab13 activation assays, reciprocal co-IP, migration/adhesion assays, and Rab13-deficient mouse model\",\n      \"pmids\": [\"25074980\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Phosphorylation site(s) on DENND1C not mapped\", \"How phosphorylation switches GEF specificity toward Rab13 unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Placing DENND1C/Rab35 downstream of the IL-17R/Act1 complex connected its GEF activity to receptor signaling driving PKCα-fascin-dependent smooth muscle contraction.\",\n      \"evidence\": \"Co-IP of recruitment to IL-17R/Act1, Rab35 knockdown with PKCα and fascin phosphorylation readouts, and contraction assays in airway smooth muscle cells\",\n      \"pmids\": [\"30683702\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single-lab finding without independent replication\", \"Direct versus indirect recruitment of DENND1C to the complex not distinguished\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Testing DENND1C against other Rab35 GEFs established that its activity is context-restricted, ruling it out as the primary GEF for epithelial podocalyxin trafficking.\",\n      \"evidence\": \"DENND1C knockout in MDCK II cells with podocalyxin localization imaging in 2D and 3D\",\n      \"pmids\": [\"31992598\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Possible redundancy with other GEFs not excluded\", \"Does not identify which contexts depend specifically on DENND1C\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How DENND1C selects between its Rab35 and Rab13 targets, and what determines its tissue- and context-specific deployment, remains unresolved.\",\n      \"evidence\": \"No single study reconciles the GEF target switch with upstream regulation\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of dual Rab35/Rab13 specificity unknown\", \"Phosphorylation sites and their effect on substrate choice unmapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RAB35\", \"RAB13\", \"STK4\", \"fascin\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}