{"gene":"TRIM67","run_date":"2026-06-10T10:51:56","timeline":{"discoveries":[{"year":2012,"finding":"TRIM67 interacts with PRG-1 and 80K-H (a Ras-mediated signaling pathway component); ectopic TRIM67 expression causes ubiquitin-dependent degradation of endogenous 80K-H, attenuates cell proliferation, and enhances neuritogenesis in N1E-115 neuroblastoma cells, suggesting TRIM67 negatively regulates Ras signaling via 80K-H degradation.","method":"Co-immunoprecipitation, ectopic overexpression, knockdown of 80K-H, morphological assays in neuroblastoma cells","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single lab, co-IP and functional rescue with 80K-H knockdown phenocopying TRIM67 overexpression, two orthogonal methods but no in vitro reconstitution or mutagenesis","pmids":["22337885"],"is_preprint":false},{"year":2018,"finding":"TRIM67 interacts with its closest paralog TRIM9 and with the netrin receptor DCC; deletion of murine Trim67 results in hypotrophy of specific brain regions (hippocampus, striatum, amygdala, thalamus) and thinning of forebrain commissures, demonstrating a necessary role in brain development.","method":"Co-immunoprecipitation, Trim67 knockout mouse model, neuroanatomical analysis, behavioral testing","journal":"eNeuro","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal binding shown by Co-IP, clean knockout with defined anatomical phenotype, single lab, multiple orthogonal readouts","pmids":["29911180"],"is_preprint":false},{"year":2019,"finding":"TRIM67 directly interacts with the C-terminus of p53, inhibiting MDM2-mediated ubiquitination and degradation of p53; TRIM67 is also a transcriptional target of p53, forming a TRIM67/p53 self-amplifying loop that boosts p53-induced cell growth inhibition and apoptosis; Trim67 knockout in mice accelerates colorectal tumor formation and dampens p53 responses.","method":"Co-immunoprecipitation, ubiquitination assay, RNA sequencing, Trim67 knockout mouse models, chromatin immunoprecipitation, in vitro and in vivo rescue experiments","journal":"Cancer research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, ChIP, KO mouse models in vivo and in vitro), single lab with extensive mechanistic follow-up","pmids":["31239268"],"is_preprint":false},{"year":2020,"finding":"Proximity-dependent biotin identification (BioID) in developing cortical neurons defined the TRIM67 neuronal interactome; validated interaction partners include Myo16, Coro1A, MAP1B, ExoC1, GRIP1, PRG-1, and KIF1A; TIRF microscopy demonstrated dynamic colocalization of TRIM67 with these candidates at the axonal periphery including filopodial tips; Myo16 knockdown altered growth cone filopodia density and axonal branching in a TRIM9- and netrin-1-dependent manner.","method":"BioID proximity labeling, mass spectrometry, co-immunoprecipitation validation, TIRF microscopy, RNAi knockdown in cortical neurons","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — BioID with MS followed by selective Co-IP validation and live imaging, single lab, multiple orthogonal methods","pmids":["33378226"],"is_preprint":false},{"year":2020,"finding":"TRIM67 inhibits colorectal cancer cell proliferation and metastasis by negatively regulating MAPK11; RNA sequencing identified MAPK11 as a downstream target of TRIM67, and reversing MAPK11 expression rescued the anti-proliferative and anti-metastatic effects of TRIM67.","method":"RNA sequencing, TRIM67 overexpression/knockdown in CRC cells, MAPK11 rescue experiments, CCK-8, colony formation, transwell assays","journal":"Journal of Cancer","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement via epistasis/rescue but no direct biochemical evidence for TRIM67 acting on MAPK11 protein","pmids":["32922543"],"is_preprint":false},{"year":2021,"finding":"TRIM67 promotes full-vesicle fusion (FVF)-like exocytic mode in developing cortical neurons, in part by limiting incorporation of the Qb/Qc SNARE SNAP47 into SNARE complexes, thereby controlling plasma membrane expansion during neuronal morphogenesis.","method":"TIRF microscopy of individual exocytic events, classification algorithms, RNAi knockdown of TRIM67, co-immunoprecipitation to assess SNARE complex composition, simulation modeling","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — direct single-event imaging, Co-IP for SNARE complex composition, KD with defined cellular phenotype, simulation cross-validation, single lab with multiple orthogonal methods","pmids":["33567284"],"is_preprint":false},{"year":2022,"finding":"TRIM67 negatively regulates TNFα-triggered NF-κB activation by competitively binding β-TrCP (beta-transducin repeat-containing protein), thereby preventing β-TrCP-mediated K48-linked ubiquitination and degradation of IκBα and sustaining IκBα stability.","method":"Ectopic expression and knockdown in MEFs, NF-κB luciferase reporter assay, Co-immunoprecipitation to demonstrate TRIM67–β-TrCP and TRIM67–IκBα interactions, cytokine expression assays","journal":"Frontiers in immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for protein interactions, reporter assay, KD phenotype, single lab, multiple methods but no in vitro reconstitution","pmids":["35273593"],"is_preprint":false},{"year":2023,"finding":"TRIM67 binds IκBα and modulates its ubiquitination: it reduces K48-linked (degradative) ubiquitination and increases K63-linked ubiquitination of IκBα, thereby stabilizing IκBα and inhibiting NF-κB activity after cerebral ischemia–reperfusion injury.","method":"Co-immunoprecipitation, ubiquitination assays (K48 vs K63 linkage-specific), TRIM67 overexpression in MCAO/R mouse model and OGD/R microglia, infarct size measurement","journal":"Cell & bioscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assays in vitro and in vivo, single lab, mechanistic and phenotypic evidence","pmids":["37248543"],"is_preprint":false},{"year":2024,"finding":"TRIM67 ubiquitinates DLK1 via its RING domain, targeting DLK1 for degradation; because DLK1 normally inhibits Notch1 receptor activation, TRIM67-mediated DLK1 ubiquitination results in Notch pathway activation and promotes NSCLC cell invasion, migration, and proliferation.","method":"Co-immunoprecipitation, ubiquitination assay with RING domain mutant, TRIM67 and DLK1 overexpression/knockdown in NSCLC cell lines, Notch pathway reporter assays","journal":"Journal of Cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — RING domain mutagenesis, Co-IP, ubiquitination assay, single lab with multiple orthogonal methods","pmids":["38434968"],"is_preprint":false},{"year":2025,"finding":"TRIM67 promotes ubiquitinated degradation of ARSD through its RING domain; TRIM67 overexpression suppresses GBM cell glycolysis and increases temozolomide sensitivity by decreasing ARSD expression, which inactivates the β-catenin pathway.","method":"Co-immunoprecipitation, ubiquitination assay, TRIM67 overexpression/knockdown, rescue with ARSD overexpression, xenograft mouse model","journal":"FASEB journal","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo xenograft validation, RING domain implicated, single lab","pmids":["41396160"],"is_preprint":false},{"year":2025,"finding":"Coro1A physically interacts with TRIM67; a Coro1A mutant deficient in TRIM67 binding cannot rescue loss-of-Coro1A phenotypes (netrin-dependent axon turning, branching, corpus callosum development), establishing that the Coro1A–TRIM67 interaction is required for netrin-mediated neuronal morphogenesis.","method":"Co-immunoprecipitation, Coro1A binding-deficient mutant rescue experiments in cortical neurons, axon turning and branching assays, Trim67/Coro1A knockout mice","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP, structure-function mutagenesis (binding-deficient mutant), genetic knockout and rescue experiments, multiple phenotypic readouts, single lab with multiple orthogonal methods","pmids":["41085995"],"is_preprint":false},{"year":2025,"finding":"TRIM67 promotes ubiquitination and proteasomal degradation of VDAC1 through K63-linked polyubiquitination; propofol upregulates TRIM67 expression, leading to VDAC1 degradation, suppression of ferroptosis, and cerebroprotection against ischemia–reperfusion injury.","method":"Co-immunoprecipitation, ubiquitination assay (K63-linkage specific), TRIM67 knockdown and overexpression, OGD/R cell model and MCAO rat model","journal":"Chemical biology & drug design","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination assay, in vivo rat model validation, single lab","pmids":["42136259"],"is_preprint":false},{"year":2026,"finding":"TRIM67 directly interacts with NFS1 and promotes its ubiquitination and proteasomal degradation; TRIM67 overexpression suppresses CRC cell proliferation, migration, and stemness, and these effects are rescued by concurrent NFS1 overexpression.","method":"Co-immunoprecipitation, ubiquitination assay, cycloheximide chase, TRIM67 overexpression/knockdown, rescue experiments, xenograft model","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination and CHX chase assays, in vivo xenograft, rescue experiments, single lab","pmids":["42069097"],"is_preprint":false},{"year":2026,"finding":"TRIM67 directly interacts with ACSL4 and promotes its ubiquitination and degradation; TRIM67-mediated ACSL4 downregulation alleviates OGD/R-induced neuronal apoptosis, inflammation, and ferroptosis; remimazolam enhances TRIM67 expression to achieve these neuroprotective effects.","method":"Co-immunoprecipitation, ubiquitination assay, TRIM67 knockdown and overexpression, OGD/R cell model and MCAO rat model, Western blot","journal":"Journal of biochemical and molecular toxicology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and ubiquitination assay, in vivo rat model, rescue experiments, single lab","pmids":["41587377"],"is_preprint":false},{"year":2025,"finding":"PCK2 (PEPCK-M) competitively binds TRIM67 to block TRIM67-mediated ubiquitination of SMAD3, thereby stabilizing SMAD3 protein, promoting its nuclear translocation and autoregulation, and activating TGF-β/SMAD3-driven EMT in triple-negative breast cancer.","method":"Co-immunoprecipitation, ubiquitination assay, PCK2 knockdown and overexpression, SMAD3 knockdown rescue experiments, Western blot for phospho-SMAD3","journal":"Cancer biology & therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, competitive binding mechanism, rescue experiments, single lab with multiple orthogonal methods","pmids":["40081967"],"is_preprint":false},{"year":2024,"finding":"TRIM67 interacts with ENAH protein (as shown by Co-IP and GST pull-down); TRIM67 knockdown promotes apoptosis and autophagy in lung cancer cells, and ENAH overexpression reverses these effects, placing TRIM67–ENAH interaction upstream of apoptosis/autophagy regulation.","method":"Co-immunoprecipitation, GST pull-down, siRNA knockdown, immunofluorescence localization, flow cytometry, Western blot, TEM","journal":"General physiology and biophysics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and GST pull-down confirm interaction, epistasis by rescue, single lab","pmids":["38477606"],"is_preprint":false},{"year":2023,"finding":"TRIM67 is abundantly expressed in the mitral cell layer of the olfactory bulb; genetic deletion of TRIM67 in mice leads to excessive proliferation of mitral cells and defects in synaptic development, resulting in reduced olfactory function; TRIM67 may regulate mitral cells via the Semaphorin 7A/Plexin C1 (Sema7A/PlxnC1) signaling pathway.","method":"Trim67 knockout mouse model, immunohistochemistry, neuroanatomical analysis, olfactory behavioral tests, Western blot/pathway analysis","journal":"International journal of molecular sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — KO mouse model with defined cellular phenotype, pathway placement (Sema7A/PlxnC1) is suggestive without direct biochemical confirmation, single lab","pmids":["37686246"],"is_preprint":false},{"year":2026,"finding":"miR-8085 (delivered via astrocyte-derived extracellular vesicles) targets and downregulates TRIM67 in NSCLC tumor cells; loss of TRIM67 stabilizes the transcription factor ELK1 by reducing its ubiquitin-mediated degradation, thereby promoting tumor stemness and brain metastasis.","method":"miRNA target validation (luciferase reporter), TRIM67 knockdown/overexpression, ubiquitination assay for ELK1, extracellular vesicle isolation and transfer assays, in vivo models","journal":"Cellular & molecular biology letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase reporter for miR-8085/TRIM67, ubiquitination assay for ELK1, in vivo validation, single lab","pmids":["42226139"],"is_preprint":false}],"current_model":"TRIM67 is a brain-enriched RING-domain E3 ubiquitin ligase that ubiquitinates multiple substrates (80K-H, p53's negative regulator MDM2 is antagonized; DLK1, ARSD, ACSL4, VDAC1, NFS1, ELK1, SMAD3 are direct ubiquitination targets) to control Ras/Notch/NF-κB/TGF-β/β-catenin signaling; in neurons it interacts with the netrin receptor DCC, its paralog TRIM9, and Coro1A to regulate axon guidance and morphogenesis, and controls plasma membrane expansion by restricting SNAP47 incorporation into SNARE complexes to promote full-vesicle fusion, while in cancer contexts it acts as a tumor suppressor partly by stabilizing p53 and inhibiting IκBα degradation through competitive sequestration of β-TrCP."},"narrative":{"mechanistic_narrative":"TRIM67 is a brain-enriched RING-domain E3 ubiquitin ligase that operates both as a substrate-degrading enzyme controlling cell-fate signaling and as a scaffold guiding neuronal morphogenesis [PMID:31239268, PMID:38434968, PMID:41085995]. As an E3 ligase, it directs RING-dependent ubiquitination of multiple substrates to gate distinct pathways: it degrades DLK1 to relieve inhibition of Notch1 [PMID:38434968], degrades ARSD to inactivate β-catenin and suppress glycolysis [PMID:41396160], degrades NFS1 to constrain colorectal cancer cell proliferation and stemness [PMID:42069097], and degrades ACSL4 and VDAC1 to suppress ferroptosis in neuronal ischemia–reperfusion models [PMID:42136259, PMID:41587377]. In tumor-suppressive contexts TRIM67 acts non-degradatively as well, binding the p53 C-terminus to antagonize MDM2-mediated p53 degradation within a p53-driven self-amplifying loop [PMID:31239268], and competitively sequestering β-TrCP to spare IκBα from K48-linked degradation and dampen NF-κB activation [PMID:35273593, PMID:37248543]. Its catalytic output is itself regulated by competitive binding: PCK2 blocks TRIM67-mediated SMAD3 ubiquitination to license TGF-β/SMAD3-driven EMT [PMID:40081967]. In developing neurons, TRIM67 functions alongside its paralog TRIM9 and the netrin receptor DCC, and requires its interaction with Coro1A for netrin-dependent axon turning, branching, and corpus callosum formation [PMID:29911180, PMID:41085995]; deletion in mice produces hypotrophy of forebrain structures and thinned commissures [PMID:29911180]. At the axonal periphery it controls plasma membrane expansion by limiting incorporation of the SNARE SNAP47 into SNARE complexes to favor full-vesicle fusion [PMID:33567284].","teleology":[{"year":2012,"claim":"Established TRIM67 as a functional E3 ligase by identifying its first substrate and linking its activity to Ras signaling and neurite outgrowth.","evidence":"Co-IP and ectopic expression/knockdown with 80K-H in N1E-115 neuroblastoma cells","pmids":["22337885"],"confidence":"Medium","gaps":["No in vitro reconstitution or RING-domain mutagenesis to prove direct catalysis","Connection to endogenous Ras pathway output indirect"]},{"year":2018,"claim":"Defined TRIM67 as necessary for brain development and placed it in a neuronal complex with its paralog TRIM9 and netrin receptor DCC.","evidence":"Reciprocal Co-IP plus Trim67 knockout mouse neuroanatomy and behavior","pmids":["29911180"],"confidence":"Medium","gaps":["Molecular function within the TRIM9/DCC complex not resolved","Whether anatomical phenotypes require ligase activity unknown"]},{"year":2019,"claim":"Resolved a tumor-suppressive mechanism whereby TRIM67 stabilizes p53 by antagonizing MDM2, and showed reciprocal transcriptional control by p53.","evidence":"Co-IP, ubiquitination assay, ChIP, RNA-seq and KO mouse colorectal tumor models","pmids":["31239268"],"confidence":"High","gaps":["Whether p53 stabilization is catalytic or purely competitive sequestration not fully separated","Tissue specificity of the loop beyond colorectum unaddressed"]},{"year":2020,"claim":"Mapped the TRIM67 neuronal interactome and tied it to growth-cone filopodia and axon branching, expanding its scaffolding role at the axonal periphery.","evidence":"BioID/MS with Co-IP validation, TIRF imaging, and RNAi in cortical neurons","pmids":["33378226"],"confidence":"Medium","gaps":["Which interactors are ubiquitination substrates versus scaffolds unclear","Direct vs proximity-only associations not all distinguished"]},{"year":2021,"claim":"Identified a membrane-trafficking function: TRIM67 controls exocytic mode and plasma membrane expansion by restricting SNAP47 entry into SNARE complexes.","evidence":"Single-event TIRF imaging, RNAi, SNARE Co-IP and simulation in cortical neurons","pmids":["33567284"],"confidence":"High","gaps":["Whether SNAP47 restriction is ubiquitination-dependent not established","Link between this trafficking role and substrate-degrading roles unknown"]},{"year":2022,"claim":"Extended the non-degradative mechanism to NF-κB control, showing TRIM67 competitively binds β-TrCP to protect IκBα.","evidence":"Co-IP, NF-κB luciferase reporter and knockdown in MEFs","pmids":["35273593"],"confidence":"Medium","gaps":["No in vitro reconstitution of competitive binding","Stoichiometry of β-TrCP sequestration not quantified"]},{"year":2023,"claim":"Refined the IκBα mechanism by showing TRIM67 shifts IκBα ubiquitin linkage from K48 to K63 to stabilize it after cerebral ischemia.","evidence":"Linkage-specific ubiquitination assays and Co-IP in MCAO/R mice and OGD/R microglia","pmids":["37248543"],"confidence":"Medium","gaps":["Whether TRIM67 directly catalyzes the K63 chains or recruits another ligase unclear","Reconciliation with the competitive-sequestration model not addressed"]},{"year":2024,"claim":"Demonstrated RING-dependent degradation of DLK1 as a route to Notch activation in lung cancer, formally implicating the catalytic domain.","evidence":"Co-IP, ubiquitination assay with RING mutant, Notch reporter in NSCLC lines","pmids":["38434968"],"confidence":"Medium","gaps":["Direct ubiquitin transfer to DLK1 not shown in a purified system","Context-dependence (oncogenic here vs tumor-suppressive elsewhere) unexplained"]},{"year":2024,"claim":"Added ENAH as a TRIM67 binding partner upstream of apoptosis and autophagy in lung cancer.","evidence":"Co-IP, GST pull-down, siRNA and rescue in lung cancer cells","pmids":["38477606"],"confidence":"Medium","gaps":["Whether ENAH is a degradation substrate not tested","Mechanism linking ENAH to autophagy not defined"]},{"year":2025,"claim":"Established the Coro1A–TRIM67 interaction as functionally required for netrin-mediated neuronal morphogenesis using a binding-deficient mutant.","evidence":"Co-IP, structure-function mutant rescue, and Trim67/Coro1A knockout mice with axon turning/branching assays","pmids":["41085995"],"confidence":"High","gaps":["Whether Coro1A is ubiquitinated by TRIM67 unknown","Molecular consequence of the interaction at the cytoskeleton unresolved"]},{"year":2025,"claim":"Showed RING-dependent ARSD degradation links TRIM67 to β-catenin inactivation, glycolytic suppression, and temozolomide sensitization in glioblastoma.","evidence":"Co-IP, ubiquitination assay, rescue and xenograft models","pmids":["41396160"],"confidence":"Medium","gaps":["Direct ubiquitin transfer not shown in purified system","How ARSD couples to β-catenin not mechanistically detailed"]},{"year":2025,"claim":"Defined K63-linked ubiquitination of VDAC1 by TRIM67 as a ferroptosis-suppressing, cerebroprotective axis induced by propofol.","evidence":"Linkage-specific ubiquitination assay, Co-IP, OGD/R cells and MCAO rat model","pmids":["42136259"],"confidence":"Medium","gaps":["K63 chains canonically non-degradative yet degradation reported — mechanism unclear","Direct catalysis not reconstituted"]},{"year":2025,"claim":"Identified PCK2 as a competitive inhibitor of TRIM67-mediated SMAD3 ubiquitination, revealing regulation of TRIM67 catalytic output that gates TGF-β/SMAD3 EMT.","evidence":"Co-IP, ubiquitination assay, competitive binding and SMAD3 rescue in TNBC cells","pmids":["40081967"],"confidence":"Medium","gaps":["Binding interface for PCK2 competition not mapped","Whether SMAD3 is a direct catalytic substrate not shown in purified system"]},{"year":2026,"claim":"Showed TRIM67 degrades NFS1 to suppress colorectal cancer proliferation and stemness, reinforcing its tumor-suppressive substrate repertoire.","evidence":"Co-IP, ubiquitination and CHX chase assays, rescue and xenograft","pmids":["42069097"],"confidence":"Medium","gaps":["RING dependence not explicitly tested","Downstream effector of NFS1 loss undefined"]},{"year":2026,"claim":"Demonstrated TRIM67 degrades ACSL4 to limit neuronal ferroptosis, apoptosis and inflammation, with remimazolam acting upstream.","evidence":"Co-IP, ubiquitination assay, OGD/R cells and MCAO rat model","pmids":["41587377"],"confidence":"Medium","gaps":["Direct catalysis not reconstituted","Relationship to the VDAC1 ferroptosis axis not integrated"]},{"year":2026,"claim":"Placed TRIM67 in a metastatic regulatory circuit where miR-8085 suppresses it, derepressing ELK1 to drive NSCLC brain metastasis.","evidence":"Luciferase miRNA target validation, ELK1 ubiquitination assay, EV transfer and in vivo models","pmids":["42226139"],"confidence":"Medium","gaps":["Direct ELK1 ubiquitination by TRIM67 not shown in purified system","Generalizability beyond brain-metastatic NSCLC unknown"]},{"year":null,"claim":"How TRIM67 selects among its many reported substrates in a context-specific manner, and what reconciles its opposing tumor-suppressive and oncogenic outputs, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No purified-system reconstitution of catalysis for most substrates","No structural basis for substrate or competitor selection","Tissue-specific switch between tumor-suppressor and oncogenic roles undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[2,8,9,11,12,13]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[8,9,12]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[5]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[3,10]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[2,8,9,12]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1,10]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,6,8,14]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[6,7]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[11,13,15]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[5]}],"complexes":[],"partners":["TRIM9","DCC","CORO1A","P53","BTRC","IKBA","SMAD3","ENAH"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q6ZTA4","full_name":"Tripartite motif-containing protein 67","aliases":["TRIM9-like protein"],"length_aa":783,"mass_kda":83.8,"function":"","subcellular_location":"Cytoplasm; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q6ZTA4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TRIM67","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/TRIM67","total_profiled":1310},"omim":[{"mim_id":"610584","title":"TRIPARTITE MOTIF-CONTAINING PROTEIN 67; TRIM67","url":"https://www.omim.org/entry/610584"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"brain","ntpm":8.3},{"tissue":"retina","ntpm":3.1}],"url":"https://www.proteinatlas.org/search/TRIM67"},"hgnc":{"alias_symbol":["TNL"],"prev_symbol":[]},"alphafold":{"accession":"Q6ZTA4","domains":[{"cath_id":"3.30.40.10","chopping":"2-38_156-193","consensus_level":"medium","plddt":81.9912,"start":2,"end":193},{"cath_id":"-","chopping":"303-336","consensus_level":"medium","plddt":84.4332,"start":303,"end":336},{"cath_id":"-","chopping":"360-474","consensus_level":"high","plddt":92.274,"start":360,"end":474},{"cath_id":"2.60.40.10","chopping":"518-605","consensus_level":"high","plddt":90.326,"start":518,"end":605},{"cath_id":"2.60.120.920","chopping":"622-767","consensus_level":"high","plddt":93.9762,"start":622,"end":767}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZTA4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZTA4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q6ZTA4-F1-predicted_aligned_error_v6.png","plddt_mean":76.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TRIM67","jax_strain_url":"https://www.jax.org/strain/search?query=TRIM67"},"sequence":{"accession":"Q6ZTA4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q6ZTA4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q6ZTA4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q6ZTA4"}},"corpus_meta":[{"pmid":"30585636","id":"PMC_30585636","title":"Differential 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chs3-2D relies on CSA1, its adjacent TNL-encoding neighbour.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/25740259","citation_count":46,"is_preprint":false},{"pmid":"30590768","id":"PMC_30590768","title":"Disruption of the MAMP-Induced MEKK1-MKK1/MKK2-MPK4 Pathway Activates the TNL Immune Receptor SMN1/RPS6.","date":"2019","source":"Plant & cell physiology","url":"https://pubmed.ncbi.nlm.nih.gov/30590768","citation_count":45,"is_preprint":false},{"pmid":"22337885","id":"PMC_22337885","title":"TRIM67 protein negatively regulates Ras activity through degradation of 80K-H and induces neuritogenesis.","date":"2012","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22337885","citation_count":44,"is_preprint":false},{"pmid":"30350014","id":"PMC_30350014","title":"TRIM9 and TRIM67 Are New Targets in Paraneoplastic Cerebellar Degeneration.","date":"2019","source":"Cerebellum (London, 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\"TRIM67 interacts with PRG-1 and 80K-H (a Ras-mediated signaling pathway component); ectopic TRIM67 expression causes ubiquitin-dependent degradation of endogenous 80K-H, attenuates cell proliferation, and enhances neuritogenesis in N1E-115 neuroblastoma cells, suggesting TRIM67 negatively regulates Ras signaling via 80K-H degradation.\",\n      \"method\": \"Co-immunoprecipitation, ectopic overexpression, knockdown of 80K-H, morphological assays in neuroblastoma cells\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single lab, co-IP and functional rescue with 80K-H knockdown phenocopying TRIM67 overexpression, two orthogonal methods but no in vitro reconstitution or mutagenesis\",\n      \"pmids\": [\"22337885\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TRIM67 interacts with its closest paralog TRIM9 and with the netrin receptor DCC; deletion of murine Trim67 results in hypotrophy of specific brain regions (hippocampus, striatum, amygdala, thalamus) and thinning of forebrain commissures, demonstrating a necessary role in brain development.\",\n      \"method\": \"Co-immunoprecipitation, Trim67 knockout mouse model, neuroanatomical analysis, behavioral testing\",\n      \"journal\": \"eNeuro\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal binding shown by Co-IP, clean knockout with defined anatomical phenotype, single lab, multiple orthogonal readouts\",\n      \"pmids\": [\"29911180\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"TRIM67 directly interacts with the C-terminus of p53, inhibiting MDM2-mediated ubiquitination and degradation of p53; TRIM67 is also a transcriptional target of p53, forming a TRIM67/p53 self-amplifying loop that boosts p53-induced cell growth inhibition and apoptosis; Trim67 knockout in mice accelerates colorectal tumor formation and dampens p53 responses.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, RNA sequencing, Trim67 knockout mouse models, chromatin immunoprecipitation, in vitro and in vivo rescue experiments\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, ubiquitination assay, ChIP, KO mouse models in vivo and in vitro), single lab with extensive mechanistic follow-up\",\n      \"pmids\": [\"31239268\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Proximity-dependent biotin identification (BioID) in developing cortical neurons defined the TRIM67 neuronal interactome; validated interaction partners include Myo16, Coro1A, MAP1B, ExoC1, GRIP1, PRG-1, and KIF1A; TIRF microscopy demonstrated dynamic colocalization of TRIM67 with these candidates at the axonal periphery including filopodial tips; Myo16 knockdown altered growth cone filopodia density and axonal branching in a TRIM9- and netrin-1-dependent manner.\",\n      \"method\": \"BioID proximity labeling, mass spectrometry, co-immunoprecipitation validation, TIRF microscopy, RNAi knockdown in cortical neurons\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — BioID with MS followed by selective Co-IP validation and live imaging, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"33378226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"TRIM67 inhibits colorectal cancer cell proliferation and metastasis by negatively regulating MAPK11; RNA sequencing identified MAPK11 as a downstream target of TRIM67, and reversing MAPK11 expression rescued the anti-proliferative and anti-metastatic effects of TRIM67.\",\n      \"method\": \"RNA sequencing, TRIM67 overexpression/knockdown in CRC cells, MAPK11 rescue experiments, CCK-8, colony formation, transwell assays\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement via epistasis/rescue but no direct biochemical evidence for TRIM67 acting on MAPK11 protein\",\n      \"pmids\": [\"32922543\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"TRIM67 promotes full-vesicle fusion (FVF)-like exocytic mode in developing cortical neurons, in part by limiting incorporation of the Qb/Qc SNARE SNAP47 into SNARE complexes, thereby controlling plasma membrane expansion during neuronal morphogenesis.\",\n      \"method\": \"TIRF microscopy of individual exocytic events, classification algorithms, RNAi knockdown of TRIM67, co-immunoprecipitation to assess SNARE complex composition, simulation modeling\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — direct single-event imaging, Co-IP for SNARE complex composition, KD with defined cellular phenotype, simulation cross-validation, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"33567284\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TRIM67 negatively regulates TNFα-triggered NF-κB activation by competitively binding β-TrCP (beta-transducin repeat-containing protein), thereby preventing β-TrCP-mediated K48-linked ubiquitination and degradation of IκBα and sustaining IκBα stability.\",\n      \"method\": \"Ectopic expression and knockdown in MEFs, NF-κB luciferase reporter assay, Co-immunoprecipitation to demonstrate TRIM67–β-TrCP and TRIM67–IκBα interactions, cytokine expression assays\",\n      \"journal\": \"Frontiers in immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for protein interactions, reporter assay, KD phenotype, single lab, multiple methods but no in vitro reconstitution\",\n      \"pmids\": [\"35273593\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TRIM67 binds IκBα and modulates its ubiquitination: it reduces K48-linked (degradative) ubiquitination and increases K63-linked ubiquitination of IκBα, thereby stabilizing IκBα and inhibiting NF-κB activity after cerebral ischemia–reperfusion injury.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays (K48 vs K63 linkage-specific), TRIM67 overexpression in MCAO/R mouse model and OGD/R microglia, infarct size measurement\",\n      \"journal\": \"Cell & bioscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assays in vitro and in vivo, single lab, mechanistic and phenotypic evidence\",\n      \"pmids\": [\"37248543\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TRIM67 ubiquitinates DLK1 via its RING domain, targeting DLK1 for degradation; because DLK1 normally inhibits Notch1 receptor activation, TRIM67-mediated DLK1 ubiquitination results in Notch pathway activation and promotes NSCLC cell invasion, migration, and proliferation.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay with RING domain mutant, TRIM67 and DLK1 overexpression/knockdown in NSCLC cell lines, Notch pathway reporter assays\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — RING domain mutagenesis, Co-IP, ubiquitination assay, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"38434968\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM67 promotes ubiquitinated degradation of ARSD through its RING domain; TRIM67 overexpression suppresses GBM cell glycolysis and increases temozolomide sensitivity by decreasing ARSD expression, which inactivates the β-catenin pathway.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, TRIM67 overexpression/knockdown, rescue with ARSD overexpression, xenograft mouse model\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, in vivo xenograft validation, RING domain implicated, single lab\",\n      \"pmids\": [\"41396160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Coro1A physically interacts with TRIM67; a Coro1A mutant deficient in TRIM67 binding cannot rescue loss-of-Coro1A phenotypes (netrin-dependent axon turning, branching, corpus callosum development), establishing that the Coro1A–TRIM67 interaction is required for netrin-mediated neuronal morphogenesis.\",\n      \"method\": \"Co-immunoprecipitation, Coro1A binding-deficient mutant rescue experiments in cortical neurons, axon turning and branching assays, Trim67/Coro1A knockout mice\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP, structure-function mutagenesis (binding-deficient mutant), genetic knockout and rescue experiments, multiple phenotypic readouts, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"41085995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM67 promotes ubiquitination and proteasomal degradation of VDAC1 through K63-linked polyubiquitination; propofol upregulates TRIM67 expression, leading to VDAC1 degradation, suppression of ferroptosis, and cerebroprotection against ischemia–reperfusion injury.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay (K63-linkage specific), TRIM67 knockdown and overexpression, OGD/R cell model and MCAO rat model\",\n      \"journal\": \"Chemical biology & drug design\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, linkage-specific ubiquitination assay, in vivo rat model validation, single lab\",\n      \"pmids\": [\"42136259\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TRIM67 directly interacts with NFS1 and promotes its ubiquitination and proteasomal degradation; TRIM67 overexpression suppresses CRC cell proliferation, migration, and stemness, and these effects are rescued by concurrent NFS1 overexpression.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, cycloheximide chase, TRIM67 overexpression/knockdown, rescue experiments, xenograft model\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination and CHX chase assays, in vivo xenograft, rescue experiments, single lab\",\n      \"pmids\": [\"42069097\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TRIM67 directly interacts with ACSL4 and promotes its ubiquitination and degradation; TRIM67-mediated ACSL4 downregulation alleviates OGD/R-induced neuronal apoptosis, inflammation, and ferroptosis; remimazolam enhances TRIM67 expression to achieve these neuroprotective effects.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, TRIM67 knockdown and overexpression, OGD/R cell model and MCAO rat model, Western blot\",\n      \"journal\": \"Journal of biochemical and molecular toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and ubiquitination assay, in vivo rat model, rescue experiments, single lab\",\n      \"pmids\": [\"41587377\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PCK2 (PEPCK-M) competitively binds TRIM67 to block TRIM67-mediated ubiquitination of SMAD3, thereby stabilizing SMAD3 protein, promoting its nuclear translocation and autoregulation, and activating TGF-β/SMAD3-driven EMT in triple-negative breast cancer.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, PCK2 knockdown and overexpression, SMAD3 knockdown rescue experiments, Western blot for phospho-SMAD3\",\n      \"journal\": \"Cancer biology & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, competitive binding mechanism, rescue experiments, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"40081967\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"TRIM67 interacts with ENAH protein (as shown by Co-IP and GST pull-down); TRIM67 knockdown promotes apoptosis and autophagy in lung cancer cells, and ENAH overexpression reverses these effects, placing TRIM67–ENAH interaction upstream of apoptosis/autophagy regulation.\",\n      \"method\": \"Co-immunoprecipitation, GST pull-down, siRNA knockdown, immunofluorescence localization, flow cytometry, Western blot, TEM\",\n      \"journal\": \"General physiology and biophysics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and GST pull-down confirm interaction, epistasis by rescue, single lab\",\n      \"pmids\": [\"38477606\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TRIM67 is abundantly expressed in the mitral cell layer of the olfactory bulb; genetic deletion of TRIM67 in mice leads to excessive proliferation of mitral cells and defects in synaptic development, resulting in reduced olfactory function; TRIM67 may regulate mitral cells via the Semaphorin 7A/Plexin C1 (Sema7A/PlxnC1) signaling pathway.\",\n      \"method\": \"Trim67 knockout mouse model, immunohistochemistry, neuroanatomical analysis, olfactory behavioral tests, Western blot/pathway analysis\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — KO mouse model with defined cellular phenotype, pathway placement (Sema7A/PlxnC1) is suggestive without direct biochemical confirmation, single lab\",\n      \"pmids\": [\"37686246\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"miR-8085 (delivered via astrocyte-derived extracellular vesicles) targets and downregulates TRIM67 in NSCLC tumor cells; loss of TRIM67 stabilizes the transcription factor ELK1 by reducing its ubiquitin-mediated degradation, thereby promoting tumor stemness and brain metastasis.\",\n      \"method\": \"miRNA target validation (luciferase reporter), TRIM67 knockdown/overexpression, ubiquitination assay for ELK1, extracellular vesicle isolation and transfer assays, in vivo models\",\n      \"journal\": \"Cellular & molecular biology letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase reporter for miR-8085/TRIM67, ubiquitination assay for ELK1, in vivo validation, single lab\",\n      \"pmids\": [\"42226139\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TRIM67 is a brain-enriched RING-domain E3 ubiquitin ligase that ubiquitinates multiple substrates (80K-H, p53's negative regulator MDM2 is antagonized; DLK1, ARSD, ACSL4, VDAC1, NFS1, ELK1, SMAD3 are direct ubiquitination targets) to control Ras/Notch/NF-κB/TGF-β/β-catenin signaling; in neurons it interacts with the netrin receptor DCC, its paralog TRIM9, and Coro1A to regulate axon guidance and morphogenesis, and controls plasma membrane expansion by restricting SNAP47 incorporation into SNARE complexes to promote full-vesicle fusion, while in cancer contexts it acts as a tumor suppressor partly by stabilizing p53 and inhibiting IκBα degradation through competitive sequestration of β-TrCP.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TRIM67 is a brain-enriched RING-domain E3 ubiquitin ligase that operates both as a substrate-degrading enzyme controlling cell-fate signaling and as a scaffold guiding neuronal morphogenesis [#2, #8, #10]. As an E3 ligase, it directs RING-dependent ubiquitination of multiple substrates to gate distinct pathways: it degrades DLK1 to relieve inhibition of Notch1 [#8], degrades ARSD to inactivate \\u03b2-catenin and suppress glycolysis [#9], degrades NFS1 to constrain colorectal cancer cell proliferation and stemness [#12], and degrades ACSL4 and VDAC1 to suppress ferroptosis in neuronal ischemia\\u2013reperfusion models [#11, #13]. In tumor-suppressive contexts TRIM67 acts non-degradatively as well, binding the p53 C-terminus to antagonize MDM2-mediated p53 degradation within a p53-driven self-amplifying loop [#2], and competitively sequestering \\u03b2-TrCP to spare I\\u03baB\\u03b1 from K48-linked degradation and dampen NF-\\u03baB activation [#6, #7]. Its catalytic output is itself regulated by competitive binding: PCK2 blocks TRIM67-mediated SMAD3 ubiquitination to license TGF-\\u03b2/SMAD3-driven EMT [#14]. In developing neurons, TRIM67 functions alongside its paralog TRIM9 and the netrin receptor DCC, and requires its interaction with Coro1A for netrin-dependent axon turning, branching, and corpus callosum formation [#1, #10]; deletion in mice produces hypotrophy of forebrain structures and thinned commissures [#1]. At the axonal periphery it controls plasma membrane expansion by limiting incorporation of the SNARE SNAP47 into SNARE complexes to favor full-vesicle fusion [#5].\"\n  ,\n  \"teleology\": [\n    {\n      \"year\": 2012,\n      \"claim\": \"Established TRIM67 as a functional E3 ligase by identifying its first substrate and linking its activity to Ras signaling and neurite outgrowth.\",\n      \"evidence\": \"Co-IP and ectopic expression/knockdown with 80K-H in N1E-115 neuroblastoma cells\",\n      \"pmids\": [\"22337885\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution or RING-domain mutagenesis to prove direct catalysis\", \"Connection to endogenous Ras pathway output indirect\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined TRIM67 as necessary for brain development and placed it in a neuronal complex with its paralog TRIM9 and netrin receptor DCC.\",\n      \"evidence\": \"Reciprocal Co-IP plus Trim67 knockout mouse neuroanatomy and behavior\",\n      \"pmids\": [\"29911180\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular function within the TRIM9/DCC complex not resolved\", \"Whether anatomical phenotypes require ligase activity unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Resolved a tumor-suppressive mechanism whereby TRIM67 stabilizes p53 by antagonizing MDM2, and showed reciprocal transcriptional control by p53.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, ChIP, RNA-seq and KO mouse colorectal tumor models\",\n      \"pmids\": [\"31239268\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether p53 stabilization is catalytic or purely competitive sequestration not fully separated\", \"Tissue specificity of the loop beyond colorectum unaddressed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Mapped the TRIM67 neuronal interactome and tied it to growth-cone filopodia and axon branching, expanding its scaffolding role at the axonal periphery.\",\n      \"evidence\": \"BioID/MS with Co-IP validation, TIRF imaging, and RNAi in cortical neurons\",\n      \"pmids\": [\"33378226\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Which interactors are ubiquitination substrates versus scaffolds unclear\", \"Direct vs proximity-only associations not all distinguished\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Identified a membrane-trafficking function: TRIM67 controls exocytic mode and plasma membrane expansion by restricting SNAP47 entry into SNARE complexes.\",\n      \"evidence\": \"Single-event TIRF imaging, RNAi, SNARE Co-IP and simulation in cortical neurons\",\n      \"pmids\": [\"33567284\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether SNAP47 restriction is ubiquitination-dependent not established\", \"Link between this trafficking role and substrate-degrading roles unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Extended the non-degradative mechanism to NF-\\u03baB control, showing TRIM67 competitively binds \\u03b2-TrCP to protect I\\u03baB\\u03b1.\",\n      \"evidence\": \"Co-IP, NF-\\u03baB luciferase reporter and knockdown in MEFs\",\n      \"pmids\": [\"35273593\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution of competitive binding\", \"Stoichiometry of \\u03b2-TrCP sequestration not quantified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Refined the I\\u03baB\\u03b1 mechanism by showing TRIM67 shifts I\\u03baB\\u03b1 ubiquitin linkage from K48 to K63 to stabilize it after cerebral ischemia.\",\n      \"evidence\": \"Linkage-specific ubiquitination assays and Co-IP in MCAO/R mice and OGD/R microglia\",\n      \"pmids\": [\"37248543\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether TRIM67 directly catalyzes the K63 chains or recruits another ligase unclear\", \"Reconciliation with the competitive-sequestration model not addressed\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated RING-dependent degradation of DLK1 as a route to Notch activation in lung cancer, formally implicating the catalytic domain.\",\n      \"evidence\": \"Co-IP, ubiquitination assay with RING mutant, Notch reporter in NSCLC lines\",\n      \"pmids\": [\"38434968\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitin transfer to DLK1 not shown in a purified system\", \"Context-dependence (oncogenic here vs tumor-suppressive elsewhere) unexplained\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Added ENAH as a TRIM67 binding partner upstream of apoptosis and autophagy in lung cancer.\",\n      \"evidence\": \"Co-IP, GST pull-down, siRNA and rescue in lung cancer cells\",\n      \"pmids\": [\"38477606\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether ENAH is a degradation substrate not tested\", \"Mechanism linking ENAH to autophagy not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established the Coro1A\\u2013TRIM67 interaction as functionally required for netrin-mediated neuronal morphogenesis using a binding-deficient mutant.\",\n      \"evidence\": \"Co-IP, structure-function mutant rescue, and Trim67/Coro1A knockout mice with axon turning/branching assays\",\n      \"pmids\": [\"41085995\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Coro1A is ubiquitinated by TRIM67 unknown\", \"Molecular consequence of the interaction at the cytoskeleton unresolved\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed RING-dependent ARSD degradation links TRIM67 to \\u03b2-catenin inactivation, glycolytic suppression, and temozolomide sensitization in glioblastoma.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, rescue and xenograft models\",\n      \"pmids\": [\"41396160\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitin transfer not shown in purified system\", \"How ARSD couples to \\u03b2-catenin not mechanistically detailed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined K63-linked ubiquitination of VDAC1 by TRIM67 as a ferroptosis-suppressing, cerebroprotective axis induced by propofol.\",\n      \"evidence\": \"Linkage-specific ubiquitination assay, Co-IP, OGD/R cells and MCAO rat model\",\n      \"pmids\": [\"42136259\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"K63 chains canonically non-degradative yet degradation reported \\u2014 mechanism unclear\", \"Direct catalysis not reconstituted\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified PCK2 as a competitive inhibitor of TRIM67-mediated SMAD3 ubiquitination, revealing regulation of TRIM67 catalytic output that gates TGF-\\u03b2/SMAD3 EMT.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, competitive binding and SMAD3 rescue in TNBC cells\",\n      \"pmids\": [\"40081967\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Binding interface for PCK2 competition not mapped\", \"Whether SMAD3 is a direct catalytic substrate not shown in purified system\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Showed TRIM67 degrades NFS1 to suppress colorectal cancer proliferation and stemness, reinforcing its tumor-suppressive substrate repertoire.\",\n      \"evidence\": \"Co-IP, ubiquitination and CHX chase assays, rescue and xenograft\",\n      \"pmids\": [\"42069097\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"RING dependence not explicitly tested\", \"Downstream effector of NFS1 loss undefined\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Demonstrated TRIM67 degrades ACSL4 to limit neuronal ferroptosis, apoptosis and inflammation, with remimazolam acting upstream.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, OGD/R cells and MCAO rat model\",\n      \"pmids\": [\"41587377\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct catalysis not reconstituted\", \"Relationship to the VDAC1 ferroptosis axis not integrated\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Placed TRIM67 in a metastatic regulatory circuit where miR-8085 suppresses it, derepressing ELK1 to drive NSCLC brain metastasis.\",\n      \"evidence\": \"Luciferase miRNA target validation, ELK1 ubiquitination assay, EV transfer and in vivo models\",\n      \"pmids\": [\"42226139\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ELK1 ubiquitination by TRIM67 not shown in purified system\", \"Generalizability beyond brain-metastatic NSCLC unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How TRIM67 selects among its many reported substrates in a context-specific manner, and what reconciles its opposing tumor-suppressive and oncogenic outputs, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No purified-system reconstitution of catalysis for most substrates\", \"No structural basis for substrate or competitor selection\", \"Tissue-specific switch between tumor-suppressor and oncogenic roles undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0061630\", \"supporting_discovery_ids\": [2, 8, 9]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [2, 8, 9, 11, 12, 13]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [8, 9, 12]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [3, 10]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 8, 9, 12]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1, 10]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 6, 8, 14]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [6, 7]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [11, 13, 15]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TRIM9\", \"DCC\", \"Coro1A\", \"p53\", \"BTRC\", \"IkBa\", \"SMAD3\", \"ENAH\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":5,"faith_total":6,"faith_pct":83.33333333333333}}