{"gene":"TBC1D14","run_date":"2026-04-28T21:42:58","timeline":{"discoveries":[{"year":2012,"finding":"TBC1D14 colocalizes and interacts with the autophagy kinase ULK1 on Rab11-positive recycling endosomes (REs). Overexpressed TBC1D14 tubulates ULK1-positive REs, impairing their function and inhibiting autophagosome formation. TBC1D14 binds activated (GTP-bound) Rab11 but does not act as a GAP for Rab11. Loss of Rab11 prevents TBC1D14-induced RE tubulation. Amino acid starvation causes TBC1D14 to relocalize from REs to the Golgi complex, releasing RE-derived membranes for autophagosome formation.","method":"Co-immunoprecipitation, overexpression/knockdown with fluorescence microscopy, transferrin recycling assays, live-cell imaging, Rab11 loss-of-function epistasis","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (Co-IP, live imaging, epistasis, KD), replicated by follow-up studies","pmids":["22613832"],"is_preprint":false},{"year":2012,"finding":"ULK1 and ATG9 localize to transferrin receptor-positive recycling endosomes, and recycling endosomal membranes (marked by transferrin) are incorporated into newly forming autophagosomes. TBC1D14 overexpression tubulates these compartments and blocks autophagosome formation at an early stage.","method":"Fluorescence microscopy, transferrin uptake/incorporation assays, overexpression of TBC1D14 with autophagy flux readout","journal":"Autophagy","confidence":"High","confidence_rationale":"Tier 2 — orthogonal methods, consistent with primary paper (PMID:22613832)","pmids":["22874560"],"is_preprint":false},{"year":2015,"finding":"TBC1D14 interacts with the TRAPPIII complex (a multi-subunit tethering complex and GEF for RAB1) via an N-terminal 103 amino acid region. Overexpression of this region inhibits both autophagy and secretory traffic. TRAPPC8, the mammalian TRAPPIII-specific subunit, is required for TBC1D14 to bind TRAPPIII and for RAB1 activation. TBC1D14 and TRAPPIII together regulate ATG9 trafficking independently of ULK1, maintaining the cycling pool of ATG9 required for autophagy initiation.","method":"Mass spectrometry interactomics, Co-immunoprecipitation, domain-mapping overexpression, ATG9 trafficking assays, knockdown epistasis, RAB1 activation assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — MS-based interactome plus reciprocal Co-IP plus functional epistasis with multiple KDs","pmids":["26711178"],"is_preprint":false},{"year":2010,"finding":"ENU-induced loss-of-function mutation in mouse Tbc1d14 results in embryonic lethality, implicating TBC1D14 in a critical developmental role and suggesting that RAB-GAP-mediated protein transport is essential for early embryogenesis.","method":"Forward genetic screen, positional cloning, complementation analysis in mouse","journal":"BMC genetics","confidence":"Medium","confidence_rationale":"Tier 2 — clean loss-of-function allele with embryonic lethal phenotype, but molecular mechanism not fully resolved","pmids":["21118569"],"is_preprint":false},{"year":2022,"finding":"TBC1D14 suppresses autophagy in HNSCC cells and negatively regulates the expression of macrophage erythroblast attacher (MAEA); MAEA overexpression reverses TBC1D14-induced autophagy suppression, placing MAEA downstream of TBC1D14 in autophagy regulation.","method":"Mass spectrometry, western blot, immunofluorescence, transmission electron microscopy, gain-of-function rescue experiments, in vivo mouse model","journal":"International journal of biological sciences","confidence":"Medium","confidence_rationale":"Tier 2/3 — multiple methods but single lab; MAEA-TBC1D14 axis is novel and lacks independent replication","pmids":["35342354"],"is_preprint":false},{"year":2025,"finding":"TBC1D14 promotes ubiquitin-mediated degradation of DDX31 via the TRIM25-mediated K63-linked polyubiquitination pathway, thereby inhibiting ribosome biogenesis and suppressing epithelial-mesenchymal transition and lymph node metastasis in HNSCC.","method":"Proteomic profiling, co-immunoprecipitation, ubiquitination assays, knockdown/overexpression with in vivo mouse metastasis model","journal":"International journal of biological macromolecules","confidence":"Medium","confidence_rationale":"Tier 2/3 — multiple in vitro and in vivo methods but single lab, no independent replication yet","pmids":["40784392"],"is_preprint":false},{"year":2020,"finding":"miR-146b-5p directly targets the 3'-UTR of Tbc1d14 (validated by luciferase reporter assay), reducing TBC1D14 expression and thereby inhibiting autophagy activation in RAW264.7 macrophages infected with Brucella melitensis Δper.","method":"Luciferase reporter assay, qRT-PCR, RNA-seq in Tbc1d14-expressing cells","journal":"BioMed research international","confidence":"Medium","confidence_rationale":"Tier 2 — luciferase reporter validates direct miRNA-3'UTR interaction, but functional link to autophagy is indirect","pmids":["32051823"],"is_preprint":false}],"current_model":"TBC1D14 is a TBC-domain-containing protein that binds activated RAB11 on recycling endosomes (without acting as its GAP), interacts with ULK1 and the TRAPPIII complex via its N-terminal region, and regulates starvation-induced autophagosome formation by controlling membrane delivery from RAB11-positive recycling endosomes and maintaining ATG9 trafficking through the early Golgi; upon starvation TBC1D14 relocates from recycling endosomes to the Golgi, permitting RE-derived membrane to contribute to autophagosome biogenesis, and additionally TBC1D14 promotes DDX31 ubiquitination-mediated degradation to suppress ribosome biogenesis and tumor metastasis."},"narrative":{"teleology":[{"year":2010,"claim":"A forward genetic screen established that TBC1D14 is essential for mouse embryonic development, raising the question of what cellular process it controls.","evidence":"ENU mutagenesis screen with positional cloning in mouse embryos","pmids":["21118569"],"confidence":"Medium","gaps":["Molecular mechanism underlying embryonic lethality not resolved","Whether the developmental requirement reflects autophagy, membrane trafficking, or another process is unknown"]},{"year":2012,"claim":"Identification of TBC1D14 as a RAB11- and ULK1-interacting protein on recycling endosomes established that recycling endosomal membranes serve as a membrane source for autophagosome biogenesis, with TBC1D14 acting as a negative regulator by sequestering these membranes when overexpressed and relocalizing to the Golgi upon starvation to permit their release.","evidence":"Co-immunoprecipitation, live-cell imaging, transferrin recycling assays, RAB11 epistasis, and starvation-induced relocalization in mammalian cell lines","pmids":["22613832","22874560"],"confidence":"High","gaps":["How starvation triggers TBC1D14 relocalization from recycling endosomes to the Golgi is unknown","Whether TBC1D14 possesses GAP activity toward any other Rab remains untested","Endogenous loss-of-function phenotype in mammalian cells not characterized at this stage"]},{"year":2015,"claim":"Discovery that TBC1D14 binds the TRAPPIII complex through its N-terminal 103-amino-acid region revealed a ULK1-independent axis by which TBC1D14 maintains ATG9 trafficking and RAB1 activation required for autophagy initiation.","evidence":"Mass spectrometry interactomics, domain-mapping overexpression, ATG9 trafficking assays, knockdown epistasis, and RAB1 activation assays","pmids":["26711178"],"confidence":"High","gaps":["Whether TRAPPIII binding and RAB11 binding are coordinated or represent separable pools of TBC1D14 is unresolved","Structural basis of TBC1D14–TRAPPIII interaction is unknown","How TBC1D14 regulates the RAB1 activation step mechanistically is not defined"]},{"year":2020,"claim":"Validation that miR-146b-5p directly targets TBC1D14 mRNA provided a post-transcriptional regulatory input linking innate immune signaling to TBC1D14-dependent autophagy.","evidence":"Luciferase reporter assay confirming direct 3′-UTR targeting in Brucella-infected macrophages","pmids":["32051823"],"confidence":"Medium","gaps":["Functional consequence of miR-146b-5p–mediated TBC1D14 reduction on autophagy flux is indirect","Relevance beyond Brucella infection context not tested"]},{"year":2022,"claim":"TBC1D14 was shown to suppress autophagy in head-and-neck squamous cell carcinoma cells through negative regulation of MAEA expression, extending TBC1D14's autophagy-regulatory role to a cancer context.","evidence":"Mass spectrometry, western blot, immunofluorescence, TEM, rescue experiments, and in vivo mouse model in HNSCC","pmids":["35342354"],"confidence":"Medium","gaps":["Mechanism by which TBC1D14 regulates MAEA expression is not defined","Single-lab finding lacking independent replication","Whether this axis operates outside HNSCC is unknown"]},{"year":2025,"claim":"TBC1D14 was found to promote TRIM25-mediated K63-linked ubiquitination and degradation of DDX31, thereby suppressing ribosome biogenesis and epithelial–mesenchymal transition in HNSCC, revealing a function beyond canonical autophagy regulation.","evidence":"Proteomic profiling, co-immunoprecipitation, ubiquitination assays, knockdown/overexpression with in vivo metastasis model","pmids":["40784392"],"confidence":"Medium","gaps":["Single-lab finding not independently replicated","Whether TBC1D14's role in DDX31 degradation is linked to its recycling-endosome or TRAPPIII functions is unknown","Direct structural or biochemical basis for TBC1D14–TRIM25 cooperation is not established"]},{"year":null,"claim":"Key open questions include the signal that triggers TBC1D14 relocalization from recycling endosomes to the Golgi upon starvation, whether TBC1D14 possesses GAP activity toward any Rab family member, and how its autophagy-regulatory and ribosome-biogenesis-regulatory functions are coordinated.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of TBC1D14 exists","Whether TBC1D14 has catalytically active GAP function toward any Rab is unresolved","Mechanism linking starvation sensing to TBC1D14 relocalization is unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,2,4]}],"localization":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[0,2]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0,1]}],"pathway":[{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[0,1,2,4]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,1,2]}],"complexes":["TRAPPIII"],"partners":["ULK1","RAB11","TRAPPC8","ATG9A","MAEA","DDX31","TRIM25"],"other_free_text":[]},"mechanistic_narrative":"TBC1D14 is a TBC-domain-containing regulator of autophagosome biogenesis that controls membrane delivery from recycling endosomes and ATG9 trafficking through the early secretory pathway. TBC1D14 resides on RAB11-positive recycling endosomes where it binds GTP-loaded RAB11 (without GAP activity toward RAB11) and colocalizes with the autophagy kinase ULK1; upon amino acid starvation it relocalizes to the Golgi, releasing recycling-endosome-derived membranes for autophagosome formation [PMID:22613832, PMID:22874560]. Through its N-terminal region TBC1D14 also engages the TRAPPIII tethering/GEF complex (via TRAPPC8), and this interaction independently maintains ATG9 cycling required for autophagy initiation by promoting RAB1 activation [PMID:26711178]. Loss-of-function mutation in mouse Tbc1d14 causes embryonic lethality, indicating an essential developmental role [PMID:21118569]."},"prefetch_data":{"uniprot":{"accession":"Q9P2M4","full_name":"TBC1 domain family member 14","aliases":[],"length_aa":693,"mass_kda":78.1,"function":"Plays a role in the regulation of starvation-induced autophagosome formation (PubMed:22613832). Together with the TRAPPIII complex, regulates a constitutive trafficking step from peripheral recycling endosomes to the early Golgi, maintaining the cycling pool of ATG9 required for initiation of autophagy","subcellular_location":"Golgi apparatus, cis-Golgi network; Golgi apparatus, trans-Golgi network","url":"https://www.uniprot.org/uniprotkb/Q9P2M4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TBC1D14","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/TBC1D14","total_profiled":1310},"omim":[{"mim_id":"621000","title":"SORTING NEXIN 18; SNX18","url":"https://www.omim.org/entry/621000"},{"mim_id":"614855","title":"TBC1 DOMAIN FAMILY, MEMBER 14; TBC1D14","url":"https://www.omim.org/entry/614855"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Golgi apparatus","reliability":"Supported"},{"location":"Vesicles","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/TBC1D14"},"hgnc":{"alias_symbol":["KIAA1322"],"prev_symbol":[]},"alphafold":{"accession":"Q9P2M4","domains":[{"cath_id":"1.10.8.270","chopping":"417-443_456-533","consensus_level":"high","plddt":92.843,"start":417,"end":533},{"cath_id":"1.10.472.80","chopping":"544-688","consensus_level":"high","plddt":94.6109,"start":544,"end":688}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P2M4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P2M4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9P2M4-F1-predicted_aligned_error_v6.png","plddt_mean":65.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TBC1D14","jax_strain_url":"https://www.jax.org/strain/search?query=TBC1D14"},"sequence":{"accession":"Q9P2M4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9P2M4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9P2M4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9P2M4"}},"corpus_meta":[{"pmid":"22613832","id":"PMC_22613832","title":"TBC1D14 regulates autophagosome formation via Rab11- and ULK1-positive recycling endosomes.","date":"2012","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/22613832","citation_count":322,"is_preprint":false},{"pmid":"26711178","id":"PMC_26711178","title":"TBC1D14 regulates autophagy via the TRAPP complex and ATG9 traffic.","date":"2015","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/26711178","citation_count":146,"is_preprint":false},{"pmid":"30957628","id":"PMC_30957628","title":"RAB2 regulates the formation of autophagosome and autolysosome in mammalian cells.","date":"2019","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/30957628","citation_count":96,"is_preprint":false},{"pmid":"19265681","id":"PMC_19265681","title":"Estradiol stimulates Akt, AMP-activated protein kinase (AMPK) and TBC1D1/4, but not glucose uptake in rat soleus.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19265681","citation_count":92,"is_preprint":false},{"pmid":"22874560","id":"PMC_22874560","title":"Recycling endosomes contribute to autophagosome formation.","date":"2012","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/22874560","citation_count":48,"is_preprint":false},{"pmid":"32727556","id":"PMC_32727556","title":"Novel candidate genes for ECT response prediction-a pilot study analyzing the DNA methylome of depressed patients receiving electroconvulsive therapy.","date":"2020","source":"Clinical epigenetics","url":"https://pubmed.ncbi.nlm.nih.gov/32727556","citation_count":25,"is_preprint":false},{"pmid":"27669114","id":"PMC_27669114","title":"Rabs and GAPs in starvation-induced autophagy.","date":"2016","source":"Small GTPases","url":"https://pubmed.ncbi.nlm.nih.gov/27669114","citation_count":24,"is_preprint":false},{"pmid":"21118569","id":"PMC_21118569","title":"High resolution mapping and positional cloning of ENU-induced mutations in the Rw region of mouse chromosome 5.","date":"2010","source":"BMC genetics","url":"https://pubmed.ncbi.nlm.nih.gov/21118569","citation_count":22,"is_preprint":false},{"pmid":"32051823","id":"PMC_32051823","title":"miR-146b-5p Plays a Critical Role in the Regulation of Autophagy in ∆per Brucella melitensis-Infected RAW264.7 Cells.","date":"2020","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/32051823","citation_count":17,"is_preprint":false},{"pmid":"28384198","id":"PMC_28384198","title":"TBC1D12 is a novel Rab11-binding protein that modulates neurite outgrowth of PC12 cells.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/28384198","citation_count":16,"is_preprint":false},{"pmid":"35342354","id":"PMC_35342354","title":"TBC1D14 inhibits autophagy to suppress lymph node metastasis in head and neck squamous cell carcinoma by downregulating macrophage erythroblast attacher.","date":"2022","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35342354","citation_count":14,"is_preprint":false},{"pmid":"27171758","id":"PMC_27171758","title":"TBC1D14 sets the TRAPP for ATG9.","date":"2016","source":"Autophagy","url":"https://pubmed.ncbi.nlm.nih.gov/27171758","citation_count":5,"is_preprint":false},{"pmid":"38668959","id":"PMC_38668959","title":"The chromosomal characteristics of spontaneous abortion and its potential associated copy number variants and genes.","date":"2024","source":"Journal of assisted reproduction and genetics","url":"https://pubmed.ncbi.nlm.nih.gov/38668959","citation_count":5,"is_preprint":false},{"pmid":"38664360","id":"PMC_38664360","title":"Molecular mechanisms underlying structural plasticity of electroconvulsive therapy in major depressive disorder.","date":"2024","source":"Brain imaging and behavior","url":"https://pubmed.ncbi.nlm.nih.gov/38664360","citation_count":4,"is_preprint":false},{"pmid":"40784392","id":"PMC_40784392","title":"TBC1D14 inhibits ribosome biogenesis to reduce lymph node metastasis in head and neck squamous cell carcinoma by mediating DDX31 ubiquitination.","date":"2025","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40784392","citation_count":0,"is_preprint":false},{"pmid":"37699782","id":"PMC_37699782","title":"Genomic associations with antibody response to an oral cholera vaccine.","date":"2023","source":"Vaccine","url":"https://pubmed.ncbi.nlm.nih.gov/37699782","citation_count":0,"is_preprint":false},{"pmid":"41482789","id":"PMC_41482789","title":"Luffa cylindrica flower extract induces apoptosis and autophagy in breast cancer cells.","date":"2026","source":"Pakistan journal of pharmaceutical sciences","url":"https://pubmed.ncbi.nlm.nih.gov/41482789","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10265,"output_tokens":1866,"usd":0.029392},"stage2":{"model":"claude-opus-4-6","input_tokens":5122,"output_tokens":2053,"usd":0.115403},"total_usd":0.144795,"stage1_batch_id":"msgbatch_01RPbVc9PDH9zQmYK5Hs6srm","stage2_batch_id":"msgbatch_01JVbVLzeKN2yVrsGp3zC8aM","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2012,\n      \"finding\": \"TBC1D14 colocalizes and interacts with the autophagy kinase ULK1 on Rab11-positive recycling endosomes (REs). Overexpressed TBC1D14 tubulates ULK1-positive REs, impairing their function and inhibiting autophagosome formation. TBC1D14 binds activated (GTP-bound) Rab11 but does not act as a GAP for Rab11. Loss of Rab11 prevents TBC1D14-induced RE tubulation. Amino acid starvation causes TBC1D14 to relocalize from REs to the Golgi complex, releasing RE-derived membranes for autophagosome formation.\",\n      \"method\": \"Co-immunoprecipitation, overexpression/knockdown with fluorescence microscopy, transferrin recycling assays, live-cell imaging, Rab11 loss-of-function epistasis\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (Co-IP, live imaging, epistasis, KD), replicated by follow-up studies\",\n      \"pmids\": [\"22613832\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"ULK1 and ATG9 localize to transferrin receptor-positive recycling endosomes, and recycling endosomal membranes (marked by transferrin) are incorporated into newly forming autophagosomes. TBC1D14 overexpression tubulates these compartments and blocks autophagosome formation at an early stage.\",\n      \"method\": \"Fluorescence microscopy, transferrin uptake/incorporation assays, overexpression of TBC1D14 with autophagy flux readout\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — orthogonal methods, consistent with primary paper (PMID:22613832)\",\n      \"pmids\": [\"22874560\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TBC1D14 interacts with the TRAPPIII complex (a multi-subunit tethering complex and GEF for RAB1) via an N-terminal 103 amino acid region. Overexpression of this region inhibits both autophagy and secretory traffic. TRAPPC8, the mammalian TRAPPIII-specific subunit, is required for TBC1D14 to bind TRAPPIII and for RAB1 activation. TBC1D14 and TRAPPIII together regulate ATG9 trafficking independently of ULK1, maintaining the cycling pool of ATG9 required for autophagy initiation.\",\n      \"method\": \"Mass spectrometry interactomics, Co-immunoprecipitation, domain-mapping overexpression, ATG9 trafficking assays, knockdown epistasis, RAB1 activation assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — MS-based interactome plus reciprocal Co-IP plus functional epistasis with multiple KDs\",\n      \"pmids\": [\"26711178\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"ENU-induced loss-of-function mutation in mouse Tbc1d14 results in embryonic lethality, implicating TBC1D14 in a critical developmental role and suggesting that RAB-GAP-mediated protein transport is essential for early embryogenesis.\",\n      \"method\": \"Forward genetic screen, positional cloning, complementation analysis in mouse\",\n      \"journal\": \"BMC genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean loss-of-function allele with embryonic lethal phenotype, but molecular mechanism not fully resolved\",\n      \"pmids\": [\"21118569\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TBC1D14 suppresses autophagy in HNSCC cells and negatively regulates the expression of macrophage erythroblast attacher (MAEA); MAEA overexpression reverses TBC1D14-induced autophagy suppression, placing MAEA downstream of TBC1D14 in autophagy regulation.\",\n      \"method\": \"Mass spectrometry, western blot, immunofluorescence, transmission electron microscopy, gain-of-function rescue experiments, in vivo mouse model\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — multiple methods but single lab; MAEA-TBC1D14 axis is novel and lacks independent replication\",\n      \"pmids\": [\"35342354\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TBC1D14 promotes ubiquitin-mediated degradation of DDX31 via the TRIM25-mediated K63-linked polyubiquitination pathway, thereby inhibiting ribosome biogenesis and suppressing epithelial-mesenchymal transition and lymph node metastasis in HNSCC.\",\n      \"method\": \"Proteomic profiling, co-immunoprecipitation, ubiquitination assays, knockdown/overexpression with in vivo mouse metastasis model\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2/3 — multiple in vitro and in vivo methods but single lab, no independent replication yet\",\n      \"pmids\": [\"40784392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"miR-146b-5p directly targets the 3'-UTR of Tbc1d14 (validated by luciferase reporter assay), reducing TBC1D14 expression and thereby inhibiting autophagy activation in RAW264.7 macrophages infected with Brucella melitensis Δper.\",\n      \"method\": \"Luciferase reporter assay, qRT-PCR, RNA-seq in Tbc1d14-expressing cells\",\n      \"journal\": \"BioMed research international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — luciferase reporter validates direct miRNA-3'UTR interaction, but functional link to autophagy is indirect\",\n      \"pmids\": [\"32051823\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TBC1D14 is a TBC-domain-containing protein that binds activated RAB11 on recycling endosomes (without acting as its GAP), interacts with ULK1 and the TRAPPIII complex via its N-terminal region, and regulates starvation-induced autophagosome formation by controlling membrane delivery from RAB11-positive recycling endosomes and maintaining ATG9 trafficking through the early Golgi; upon starvation TBC1D14 relocates from recycling endosomes to the Golgi, permitting RE-derived membrane to contribute to autophagosome biogenesis, and additionally TBC1D14 promotes DDX31 ubiquitination-mediated degradation to suppress ribosome biogenesis and tumor metastasis.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"TBC1D14 is a TBC-domain-containing regulator of autophagosome biogenesis that controls membrane delivery from recycling endosomes and ATG9 trafficking through the early secretory pathway. TBC1D14 resides on RAB11-positive recycling endosomes where it binds GTP-loaded RAB11 (without GAP activity toward RAB11) and colocalizes with the autophagy kinase ULK1; upon amino acid starvation it relocalizes to the Golgi, releasing recycling-endosome-derived membranes for autophagosome formation [PMID:22613832, PMID:22874560]. Through its N-terminal region TBC1D14 also engages the TRAPPIII tethering/GEF complex (via TRAPPC8), and this interaction independently maintains ATG9 cycling required for autophagy initiation by promoting RAB1 activation [PMID:26711178]. Loss-of-function mutation in mouse Tbc1d14 causes embryonic lethality, indicating an essential developmental role [PMID:21118569].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"A forward genetic screen established that TBC1D14 is essential for mouse embryonic development, raising the question of what cellular process it controls.\",\n      \"evidence\": \"ENU mutagenesis screen with positional cloning in mouse embryos\",\n      \"pmids\": [\"21118569\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism underlying embryonic lethality not resolved\",\n        \"Whether the developmental requirement reflects autophagy, membrane trafficking, or another process is unknown\"\n      ]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identification of TBC1D14 as a RAB11- and ULK1-interacting protein on recycling endosomes established that recycling endosomal membranes serve as a membrane source for autophagosome biogenesis, with TBC1D14 acting as a negative regulator by sequestering these membranes when overexpressed and relocalizing to the Golgi upon starvation to permit their release.\",\n      \"evidence\": \"Co-immunoprecipitation, live-cell imaging, transferrin recycling assays, RAB11 epistasis, and starvation-induced relocalization in mammalian cell lines\",\n      \"pmids\": [\"22613832\", \"22874560\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"How starvation triggers TBC1D14 relocalization from recycling endosomes to the Golgi is unknown\",\n        \"Whether TBC1D14 possesses GAP activity toward any other Rab remains untested\",\n        \"Endogenous loss-of-function phenotype in mammalian cells not characterized at this stage\"\n      ]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Discovery that TBC1D14 binds the TRAPPIII complex through its N-terminal 103-amino-acid region revealed a ULK1-independent axis by which TBC1D14 maintains ATG9 trafficking and RAB1 activation required for autophagy initiation.\",\n      \"evidence\": \"Mass spectrometry interactomics, domain-mapping overexpression, ATG9 trafficking assays, knockdown epistasis, and RAB1 activation assays\",\n      \"pmids\": [\"26711178\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether TRAPPIII binding and RAB11 binding are coordinated or represent separable pools of TBC1D14 is unresolved\",\n        \"Structural basis of TBC1D14–TRAPPIII interaction is unknown\",\n        \"How TBC1D14 regulates the RAB1 activation step mechanistically is not defined\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Validation that miR-146b-5p directly targets TBC1D14 mRNA provided a post-transcriptional regulatory input linking innate immune signaling to TBC1D14-dependent autophagy.\",\n      \"evidence\": \"Luciferase reporter assay confirming direct 3′-UTR targeting in Brucella-infected macrophages\",\n      \"pmids\": [\"32051823\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional consequence of miR-146b-5p–mediated TBC1D14 reduction on autophagy flux is indirect\",\n        \"Relevance beyond Brucella infection context not tested\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"TBC1D14 was shown to suppress autophagy in head-and-neck squamous cell carcinoma cells through negative regulation of MAEA expression, extending TBC1D14's autophagy-regulatory role to a cancer context.\",\n      \"evidence\": \"Mass spectrometry, western blot, immunofluorescence, TEM, rescue experiments, and in vivo mouse model in HNSCC\",\n      \"pmids\": [\"35342354\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which TBC1D14 regulates MAEA expression is not defined\",\n        \"Single-lab finding lacking independent replication\",\n        \"Whether this axis operates outside HNSCC is unknown\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"TBC1D14 was found to promote TRIM25-mediated K63-linked ubiquitination and degradation of DDX31, thereby suppressing ribosome biogenesis and epithelial–mesenchymal transition in HNSCC, revealing a function beyond canonical autophagy regulation.\",\n      \"evidence\": \"Proteomic profiling, co-immunoprecipitation, ubiquitination assays, knockdown/overexpression with in vivo metastasis model\",\n      \"pmids\": [\"40784392\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single-lab finding not independently replicated\",\n        \"Whether TBC1D14's role in DDX31 degradation is linked to its recycling-endosome or TRAPPIII functions is unknown\",\n        \"Direct structural or biochemical basis for TBC1D14–TRIM25 cooperation is not established\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key open questions include the signal that triggers TBC1D14 relocalization from recycling endosomes to the Golgi upon starvation, whether TBC1D14 possesses GAP activity toward any Rab family member, and how its autophagy-regulatory and ribosome-biogenesis-regulatory functions are coordinated.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No structural model of TBC1D14 exists\",\n        \"Whether TBC1D14 has catalytically active GAP function toward any Rab is unresolved\",\n        \"Mechanism linking starvation sensing to TBC1D14 relocalization is unknown\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 2, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0, 1, 2, 4]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 1, 2]}\n    ],\n    \"complexes\": [\n      \"TRAPPIII\"\n    ],\n    \"partners\": [\n      \"ULK1\",\n      \"RAB11\",\n      \"TRAPPC8\",\n      \"ATG9A\",\n      \"MAEA\",\n      \"DDX31\",\n      \"TRIM25\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}