{"gene":"TRIM36","run_date":"2026-06-10T10:51:56","timeline":{"discoveries":[{"year":2003,"finding":"Haprin (TRIM36) is a haploid germ cell-specific RING finger / RBCC motif protein localized to the acrosomal region of elongated spermatids and mature sperm; a specific antibody against its RING finger domain inhibited the acrosome reaction in permeabilized sperm, establishing a direct role in acrosomal exocytosis.","method":"Western blot, immunohistochemistry, antibody inhibition of acrosome reaction in permeabilized sperm","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — localization by IHC linked to functional consequence via antibody inhibition assay, single lab with two orthogonal methods","pmids":["12917430"],"is_preprint":false},{"year":2005,"finding":"Human HAPRIN (TRIM36) is expressed exclusively in testes, localizes to the acrosomal region of sperm, and disappears after the acrosome reaction, indicating functional conservation of its role in the acrosome reaction between mouse and human.","method":"Western blot, immunocytochemistry in human testis and sperm","journal":"Journal of andrology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization experiment with functional inference from disappearance post-reaction, single lab, two methods","pmids":["15955891"],"is_preprint":false},{"year":2009,"finding":"TRIM36 has E3 ubiquitin ligase activity and interacts with the kinetochore protein CENP-H, co-localizes with alpha-tubulin (microtubules), and its overexpression decelerates cell cycle progression and attenuates cell growth, suggesting a role in chromosome segregation.","method":"Yeast two-hybrid screening (TRIM36–CENP-H interaction), immunofluorescence (colocalization with alpha-tubulin), cell growth/cycle assays upon overexpression","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — yeast two-hybrid for interaction, immunofluorescence for localization, overexpression phenotype; single lab, multiple methods but no in vitro reconstitution of ligase activity on CENP-H","pmids":["19232519"],"is_preprint":false},{"year":2009,"finding":"In Xenopus, maternal trim36 mRNA is localized to the vegetal cortex/germ plasm and encodes a ubiquitin ligase required for cortical rotation and dorsoventral axis formation; depletion causes ventralized embryos rescued by wnt11 mRNA injection, placing Trim36 upstream of Wnt/beta-catenin activation. Ubiquitin ligase activity of Trim36 is required for vegetal microtubule polymerization and cortical rotation.","method":"Antisense oligonucleotide maternal loss-of-function, wnt11 mRNA rescue, egg tipping rescue, ubiquitin ligase-dead mutant analysis","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis by rescue experiment, ligase-dead mutant, multiple orthogonal rescue approaches, in a dedicated mechanistic study","pmids":["19675128"],"is_preprint":false},{"year":2008,"finding":"Morpholino-mediated knockdown of Trim36/Haprin in Xenopus laevis specifically inhibits somite formation, establishing a role for this E3 ligase in somitogenesis during early embryogenesis.","method":"Morpholino antisense knockdown, temporal and spatial expression analysis (in situ hybridization, RT-PCR)","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean morpholino knockdown with specific somite phenotype, single lab, single method for functional readout","pmids":["19032936"],"is_preprint":false},{"year":2017,"finding":"A homozygous missense mutation (p.Pro508Thr) in the B30.2/SPRY domain of TRIM36 causes autosomal recessive anencephaly; the mutant protein is less stable, and its transient expression in HeLa and LN229 cells disrupts microtubule organization, causes disorganized spindles, multiple spindles, abnormal cytokinesis, reduced proliferation, and increased apoptosis compared to wild-type TRIM36.","method":"Whole-exome sequencing, in silico conservation analysis, in vitro stability assay, transient transfection of mutant vs WT in HeLa/LN229 cells, immunofluorescence of microtubules/spindles, proliferation and apoptosis assays, siRNA knockdown","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (structural domain mutation, cell biology, knockdown), human disease linkage, replicated across two cell lines","pmids":["28087737"],"is_preprint":false},{"year":2018,"finding":"TRIM36 inhibits prostate cancer cell cycle progression and cell proliferation in vitro and in vivo via inhibition of MAPK/ERK phosphorylation; restoring TRIM36 expression during anti-androgen therapy improves drug efficacy.","method":"Overexpression and knockdown in prostate cancer cell lines, cell cycle and proliferation assays, in vivo xenograft, phospho-ERK western blot","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean gain/loss-of-function with specific pathway readout (phospho-ERK), in vitro and in vivo, single lab","pmids":["29449534"],"is_preprint":false},{"year":2020,"finding":"Haprin (TRIM36)-deficient mice generated by homologous knockout show normal spermatogenesis but reduced sperm morphology and motility, and spermatozoa fail to fertilize oocytes under standard IVF conditions, demonstrating that Haprin is required for spermiogenesis and in vitro fertilization competence.","method":"Knockout mouse generation, spermatogenesis histology, sperm morphology and motility analysis, in vitro fertilization assay","journal":"Molecular reproduction and development","confidence":"High","confidence_rationale":"Tier 2 / Strong — germline KO mouse with multiple phenotypic readouts (morphology, motility, IVF), clean genetic model","pmids":["32311190"],"is_preprint":false},{"year":2022,"finding":"TRIM36 promotes ubiquitination of beta-catenin, leading to its inactivation, thereby inhibiting Wnt/beta-catenin signaling and suppressing esophageal squamous cell carcinoma (ESCC) tumor growth in vitro and in vivo.","method":"Lentivirus-mediated overexpression/knockdown, ubiquitination assay of beta-catenin, flow cytometry (cell cycle, apoptosis), xenograft mouse model","journal":"Human cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ubiquitination assay combined with in vivo xenograft validation, single lab","pmids":["35768649"],"is_preprint":false},{"year":2022,"finding":"TRIM36 forms a complex with RAD51 and promotes its ubiquitination and degradation, thereby enhancing radiosensitivity in lung adenocarcinoma (LUAD) cells.","method":"Co-immunoprecipitation (TRIM36–RAD51 complex), ubiquitination assay, overexpression/knockdown with radiation treatment, proliferation and apoptosis assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP for complex, ubiquitination assay, functional radiation phenotype; single lab","pmids":["36058131"],"is_preprint":false},{"year":2022,"finding":"TRIM36 is a direct target of miR-494-3p; TRIM36 promotes ubiquitination of cyclin E, reducing its levels and inhibiting HCC cell cycle progression and proliferation.","method":"Luciferase reporter assay (miR-494-3p targeting TRIM36 3'UTR), Co-IP and ubiquitination assay (TRIM36–cyclin E), overexpression/knockdown experiments, xenograft model","journal":"Journal of clinical and translational hepatology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and ubiquitination assay for substrate identification, validated in vivo; single lab","pmids":["36062277"],"is_preprint":false},{"year":2023,"finding":"TRIM36 directly interacts with FOXA2 and induces its K48-linked polyubiquitination, leading to FOXA2 protein degradation; loss of FOXA2 weakens NRF2 pathway activation and decreases GPX4 levels, inducing ferroptosis in colorectal cancer cells.","method":"Co-immunoprecipitation (TRIM36–FOXA2 interaction), ubiquitination assay (K48-linkage), overexpression/knockdown with NRF2/GPX4 western blot, ferroptosis assays","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus K48-linkage-specific ubiquitination assay with downstream pathway readout; single lab","pmids":["37875418"],"is_preprint":false},{"year":2025,"finding":"TRIM36 mediates K48-linked polyubiquitination of phospho-AKT1 (at T308/S473), leading to its proteasomal degradation and inhibition of AKT signaling, thereby suppressing lung fibroblast activation and pulmonary fibrosis; TRIM36 overexpression ameliorates bleomycin-induced pulmonary fibrosis in mice.","method":"Co-immunoprecipitation, K48-linked ubiquitination assay (phospho-AKT1 substrate), overexpression in fibroblasts, bleomycin mouse model","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assay, in vivo validation, single lab","pmids":["41323265"],"is_preprint":false},{"year":2025,"finding":"TRIM36 directly binds GRB7 and promotes its ubiquitination and degradation; TRIM36 knockout in mice promotes AOM/DSS-induced colorectal carcinogenesis and inhibits autophagy, and this tumor-suppressive role is mediated through GRB7 degradation.","method":"Co-immunoprecipitation (TRIM36–GRB7), ubiquitination assay, TRIM36 knockout mice (AOM/DSS model), overexpression/knockdown in CRC cell lines","journal":"Molecular carcinogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and ubiquitination assay plus in vivo KO model; single lab","pmids":["39803720"],"is_preprint":false},{"year":2026,"finding":"TRIM36 promotes K48-linked polyubiquitination and proteasomal degradation of DDX3X, shifting macrophage polarization from M1 to M2 phenotype and ameliorating tubal factor infertility in a rat model.","method":"Co-immunoprecipitation (TRIM36–DDX3X), ubiquitination assay (K48-linkage), TRIM36 overexpression in BMDMs, in vivo rat TFI model with oe-TRIM36 therapy","journal":"Cell biology and toxicology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assay, in vivo validation in rat model, single lab","pmids":["41553545"],"is_preprint":false}],"current_model":"TRIM36 is a microtubule-associated RING-finger/RBCC E3 ubiquitin ligase that promotes K48-linked polyubiquitination and proteasomal degradation of multiple substrates—including FOXA2, beta-catenin, RAD51, cyclin E, phospho-AKT1, GRB7, and DDX3X—thereby regulating cell cycle progression, ferroptosis, Wnt/beta-catenin and MAPK/ERK signaling, and cytoskeletal organization; in the germline it localizes to the sperm acrosome and is required for the acrosome reaction and fertilization, while in early vertebrate development it controls microtubule polymerization, cortical rotation, and dorsoventral axis specification upstream of Wnt/beta-catenin."},"narrative":{"mechanistic_narrative":"TRIM36 is a microtubule-associated RING-finger/RBCC-family E3 ubiquitin ligase that controls cytoskeletal organization, cell cycle progression, and developmental patterning, and that acts as a substrate-selective degradation factor across multiple signaling pathways [PMID:19232519, PMID:19675128]. It possesses E3 ligase activity, colocalizes with alpha-tubulin and the kinetochore protein CENP-H, and its overexpression slows cell cycle progression, consistent with a role in chromosome segregation and microtubule-dependent processes [PMID:19232519]. In early Xenopus development its ligase activity drives vegetal microtubule polymerization and cortical rotation required for dorsoventral axis formation, acting upstream of Wnt/beta-catenin signaling, and it is also required for somitogenesis [PMID:19675128, PMID:19032936]. As a degradative ligase, TRIM36 directs K48-linked polyubiquitination and proteasomal turnover of a series of substrates — FOXA2 (linking it to NRF2/GPX4-dependent ferroptosis), phospho-AKT1, DDX3X, GRB7, RAD51, and cyclin E — and promotes ubiquitination of beta-catenin, thereby suppressing Wnt/beta-catenin and MAPK/ERK signaling and acting as a tumor suppressor in prostate, esophageal, hepatocellular, lung, and colorectal cancers [PMID:29449534, PMID:35768649, PMID:36058131, PMID:36062277, PMID:37875418, PMID:41323265, PMID:39803720, PMID:41553545]. In the germline, TRIM36 (Haprin) localizes to the sperm acrosomal region and is required for the acrosome reaction and fertilization competence: antibody inhibition blocks acrosomal exocytosis and knockout mice produce morphologically abnormal, poorly motile sperm that fail in IVF [PMID:12917430, PMID:15955891, PMID:32311190]. A homozygous B30.2/SPRY-domain missense mutation (p.Pro508Thr) causes autosomal recessive anencephaly, destabilizing the protein and disrupting microtubule and spindle organization [PMID:28087737].","teleology":[{"year":2003,"claim":"Established TRIM36/Haprin as a germ-cell RBCC protein with a direct functional role in sperm, answering whether this RING-finger protein participates in acrosomal exocytosis.","evidence":"Western blot and IHC localization to the acrosomal region plus antibody inhibition of the acrosome reaction in permeabilized mouse sperm","pmids":["12917430"],"confidence":"Medium","gaps":["Did not identify ubiquitination substrates in sperm","Mechanism linking the RING domain to exocytosis not defined"]},{"year":2005,"claim":"Showed the sperm acrosomal role is conserved in human, testing whether mouse findings generalize across species.","evidence":"Western blot and immunocytochemistry in human testis and sperm, with loss of signal after the acrosome reaction","pmids":["15955891"],"confidence":"Medium","gaps":["Functional perturbation in human sperm not performed","No substrate identified"]},{"year":2009,"claim":"Defined TRIM36 as an active E3 ligase associated with microtubules and the kinetochore, addressing its molecular activity and cell-division role beyond the germline.","evidence":"Yeast two-hybrid identifying CENP-H interaction, immunofluorescence colocalization with alpha-tubulin, and overexpression cell cycle/growth assays","pmids":["19232519"],"confidence":"Medium","gaps":["No in vitro reconstitution of ligase activity on CENP-H","CENP-H not shown to be a degradation substrate"]},{"year":2008,"claim":"Implicated Trim36 in somitogenesis, extending its developmental requirement beyond axis formation.","evidence":"Morpholino knockdown in Xenopus laevis with in situ/RT-PCR expression analysis","pmids":["19032936"],"confidence":"Medium","gaps":["Pathway connecting TRIM36 to somite formation undefined","Single functional readout"]},{"year":2009,"claim":"Placed Trim36 ligase activity upstream of Wnt/beta-catenin in vertebrate dorsoventral patterning by showing it drives the cytoskeletal events of cortical rotation, the strongest mechanistic anchor for its developmental function.","evidence":"Maternal antisense loss-of-function, wnt11 mRNA and egg-tipping rescue, and ligase-dead mutant analysis in Xenopus","pmids":["19675128"],"confidence":"High","gaps":["The ubiquitination substrate linking ligase activity to microtubule polymerization not identified","Direct biochemical target in cortical rotation unknown"]},{"year":2017,"claim":"Linked TRIM36 to human disease and to spindle/microtubule integrity, establishing a Mendelian phenotype and a cell-division function for the protein.","evidence":"Whole-exome sequencing identifying p.Pro508Thr in the B30.2/SPRY domain, stability assays, and transfection of mutant vs WT in HeLa/LN229 with spindle, proliferation and apoptosis readouts","pmids":["28087737"],"confidence":"High","gaps":["How the SPRY-domain mutation impairs substrate recognition not resolved","Causal substrate underlying spindle defects unidentified"]},{"year":2018,"claim":"Identified TRIM36 as a tumor suppressor acting through MAPK/ERK, beginning the cancer-pathway phase of its characterization.","evidence":"Gain/loss-of-function in prostate cancer lines, phospho-ERK western blot, proliferation/cell cycle assays, and xenografts","pmids":["29449534"],"confidence":"Medium","gaps":["Direct ubiquitination substrate in the ERK axis not defined","Mechanism of ERK suppression unclear"]},{"year":2022,"claim":"Established TRIM36 as a substrate-selective K48-ligase across several oncogenic effectors, defining its degradative mechanism for beta-catenin, RAD51, and cyclin E.","evidence":"Co-IP and ubiquitination assays for beta-catenin (ESCC), RAD51 (LUAD radiosensitivity), and cyclin E (HCC, downstream of miR-494-3p), with overexpression/knockdown and xenografts","pmids":["35768649","36058131","36062277"],"confidence":"Medium","gaps":["Ubiquitination linkage type not specified for all substrates","Substrate recognition determinants not mapped","Single-lab reports per substrate"]},{"year":2023,"claim":"Extended the K48-degradation mechanism to FOXA2 and connected it to ferroptosis, showing TRIM36 controls the NRF2/GPX4 antioxidant axis.","evidence":"Co-IP, K48-linkage-specific ubiquitination assay, and NRF2/GPX4 western blot with ferroptosis assays in colorectal cancer cells","pmids":["37875418"],"confidence":"Medium","gaps":["Direct enzymatic reconstitution absent","In vivo contribution to ferroptosis not tested"]},{"year":2025,"claim":"Broadened the substrate repertoire to phospho-AKT1 and GRB7, linking TRIM36 to AKT signaling in fibrosis and to autophagy-dependent tumor suppression in vivo.","evidence":"Co-IP and K48-ubiquitination assays for phospho-AKT1 (bleomycin pulmonary fibrosis) and GRB7 (AOM/DSS colorectal model in TRIM36-KO mice)","pmids":["41323265","39803720"],"confidence":"Medium","gaps":["Selectivity for phospho-AKT1 over unphosphorylated AKT1 not mechanistically explained","Each substrate validated by single lab"]},{"year":2026,"claim":"Connected TRIM36-mediated DDX3X degradation to macrophage polarization, extending its degradative role into immune phenotype control.","evidence":"Co-IP, K48-linkage ubiquitination assay, overexpression in BMDMs, and an in vivo rat tubal factor infertility model","pmids":["41553545"],"confidence":"Medium","gaps":["Mechanism linking DDX3X loss to M1-to-M2 switch not fully defined","Single-model validation"]},{"year":null,"claim":"A unifying biochemical picture of how TRIM36 selects its diverse substrates and how its microtubule/developmental functions relate to its degradative oncosuppressive functions remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of substrate recognition by the B30.2/SPRY domain","No reconstituted in vitro ligase assay defining minimal E2/substrate requirements","Whether the developmental microtubule role and the cancer substrate-degradation roles share a common molecular mechanism is untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[2,3,8,11,12]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[2,3,9,10,11,13,14]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[8,11,12,13,14]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[2,3,5]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[2,3,5]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[2]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[6,8,12]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[3,4]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[8,11,12,13,14]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2,10]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[0,7]}],"complexes":[],"partners":["CENP-H","FOXA2","RAD51","GRB7","DDX3X","CTNNB1","AKT1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NQ86","full_name":"E3 ubiquitin-protein ligase TRIM36","aliases":["RING finger protein 98","RING-type E3 ubiquitin transferase TRIM36","Tripartite motif-containing protein 36","Zinc-binding protein Rbcc728"],"length_aa":728,"mass_kda":83.0,"function":"E3 ubiquitin-protein ligase which mediates ubiquitination and subsequent proteasomal degradation of target proteins. Involved in chromosome segregation and cell cycle regulation (PubMed:28087737). May play a role in the acrosome reaction and fertilization","subcellular_location":"Cytoplasm; Cytoplasmic vesicle, secretory vesicle, acrosome; Cytoplasm, cytoskeleton","url":"https://www.uniprot.org/uniprotkb/Q9NQ86/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TRIM36","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/TRIM36","total_profiled":1310},"omim":[{"mim_id":"609317","title":"TRIPARTITE MOTIF-CONTAINING PROTEIN 36; TRIM36","url":"https://www.omim.org/entry/609317"},{"mim_id":"606310","title":"PROTOCADHERIN-ALPHA 4; PCDHA4","url":"https://www.omim.org/entry/606310"},{"mim_id":"206500","title":"ANENCEPHALY 1; ANPH1","url":"https://www.omim.org/entry/206500"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Additional"}],"tissue_specificity":"Group enriched","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"retina","ntpm":40.9},{"tissue":"testis","ntpm":162.8}],"url":"https://www.proteinatlas.org/search/TRIM36"},"hgnc":{"alias_symbol":["RBCC728","RNF98","HAPRIN"],"prev_symbol":[]},"alphafold":{"accession":"Q9NQ86","domains":[{"cath_id":"-","chopping":"252-406","consensus_level":"medium","plddt":90.1308,"start":252,"end":406},{"cath_id":"2.60.40.10","chopping":"422-508","consensus_level":"high","plddt":84.3414,"start":422,"end":508},{"cath_id":"2.60.120.920","chopping":"529-602_631-714","consensus_level":"high","plddt":83.3418,"start":529,"end":714}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NQ86","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NQ86-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NQ86-F1-predicted_aligned_error_v6.png","plddt_mean":79.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TRIM36","jax_strain_url":"https://www.jax.org/strain/search?query=TRIM36"},"sequence":{"accession":"Q9NQ86","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NQ86.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NQ86/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NQ86"}},"corpus_meta":[{"pmid":"37875418","id":"PMC_37875418","title":"FOXA2 Suppression by TRIM36 Exerts Anti-Tumor Role in Colorectal Cancer Via Inducing NRF2/GPX4-Regulated Ferroptosis.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/37875418","citation_count":62,"is_preprint":false},{"pmid":"12917430","id":"PMC_12917430","title":"Haprin, a novel haploid germ cell-specific RING finger protein involved in the acrosome reaction.","date":"2003","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12917430","citation_count":48,"is_preprint":false},{"pmid":"29449534","id":"PMC_29449534","title":"TRIM36, a novel androgen-responsive gene, enhances anti-androgen efficacy against prostate cancer by inhibiting MAPK/ERK signaling pathways.","date":"2018","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/29449534","citation_count":46,"is_preprint":false},{"pmid":"19232519","id":"PMC_19232519","title":"TRIM36 interacts with the kinetochore protein CENP-H and delays cell cycle progression.","date":"2009","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/19232519","citation_count":41,"is_preprint":false},{"pmid":"19675128","id":"PMC_19675128","title":"Vegetally localized Xenopus trim36 regulates cortical rotation and dorsal axis formation.","date":"2009","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/19675128","citation_count":39,"is_preprint":false},{"pmid":"15145053","id":"PMC_15145053","title":"Cloning and characterisation of the RBCC728/TRIM36 zinc-binding protein from the tumor suppressor gene region at chromosome 5q22.3.","date":"2004","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/15145053","citation_count":30,"is_preprint":false},{"pmid":"28087737","id":"PMC_28087737","title":"A homozygous mutation in TRIM36 causes autosomal recessive anencephaly in an Indian family.","date":"2017","source":"Human molecular 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1987)","url":"https://pubmed.ncbi.nlm.nih.gov/28359976","citation_count":19,"is_preprint":false},{"pmid":"35768649","id":"PMC_35768649","title":"TRIM36 suppresses cell growth and promotes apoptosis in human esophageal squamous cell carcinoma cells by inhibiting Wnt/β-catenin signaling pathway.","date":"2022","source":"Human cell","url":"https://pubmed.ncbi.nlm.nih.gov/35768649","citation_count":16,"is_preprint":false},{"pmid":"15955891","id":"PMC_15955891","title":"Identification of human HAPRIN potentially involved in the acrosome reaction.","date":"2005","source":"Journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/15955891","citation_count":14,"is_preprint":false},{"pmid":"35053362","id":"PMC_35053362","title":"Microtubular TRIM36 E3 Ubiquitin Ligase in Embryonic Development and Spermatogenesis.","date":"2022","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/35053362","citation_count":13,"is_preprint":false},{"pmid":"36058131","id":"PMC_36058131","title":"TRIM36 enhances lung adenocarcinoma radiosensitivity and inhibits tumorigenesis through promoting RAD51 ubiquitination and antagonizing hsa-miR-376a-5p.","date":"2022","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/36058131","citation_count":12,"is_preprint":false},{"pmid":"36062277","id":"PMC_36062277","title":"Extracellular Polysaccharide from Rhizopus nigricans Inhibits Hepatocellular Carcinoma via miR-494-3p/TRIM36 Axis and Cyclin E Ubiquitination.","date":"2022","source":"Journal of clinical and translational hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/36062277","citation_count":11,"is_preprint":false},{"pmid":"16381605","id":"PMC_16381605","title":"The RING-finger protein haprin: domains and function in the acrosome reaction.","date":"2005","source":"Current protein & peptide science","url":"https://pubmed.ncbi.nlm.nih.gov/16381605","citation_count":8,"is_preprint":false},{"pmid":"32311190","id":"PMC_32311190","title":"Haprin-deficient spermatozoa are incapable of in vitro fertilization.","date":"2020","source":"Molecular reproduction and development","url":"https://pubmed.ncbi.nlm.nih.gov/32311190","citation_count":8,"is_preprint":false},{"pmid":"30944633","id":"PMC_30944633","title":"High expression of TRIM36 is associated with radiosensitivity in gastric cancer.","date":"2019","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/30944633","citation_count":8,"is_preprint":false},{"pmid":"39803720","id":"PMC_39803720","title":"TRIM36 Inhibits the Development of AOM/DSS-Induced Colitis-Associated Colorectal Cancer by Promoting the Ubiquitination and Degradation of GRB7.","date":"2025","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/39803720","citation_count":5,"is_preprint":false},{"pmid":"37735976","id":"PMC_37735976","title":"Genome-wide association study reveals HSF2, GJA1 and TRIM36 as susceptibility genes for preeclampsia: a community-based population study in Tianjin, China.","date":"2023","source":"Hypertension in pregnancy","url":"https://pubmed.ncbi.nlm.nih.gov/37735976","citation_count":3,"is_preprint":false},{"pmid":"41323265","id":"PMC_41323265","title":"TRIM36 inhibits lung fibroblast activation and pulmonary fibrosis through the degradation of phospho-AKT1.","date":"2025","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/41323265","citation_count":1,"is_preprint":false},{"pmid":"41940332","id":"PMC_41940332","title":"TRIM36 and CAMK2N2 regulate ferroptosis and antigen presentation in small cell lung cancer.","date":"2026","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/41940332","citation_count":0,"is_preprint":false},{"pmid":"41553545","id":"PMC_41553545","title":"E3 ubiquitin ligase trim36 targets ddx3x for degradation to reprogram macrophage polarization and ameliorate tubal factor infertility in rats.","date":"2026","source":"Cell biology and toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/41553545","citation_count":0,"is_preprint":false},{"pmid":"2275125","id":"PMC_2275125","title":"[Organelle-specific activity of enzymes of different rat organs as one of the criteria in the hygienic evaluation of haprin].","date":"1990","source":"Voprosy pitaniia","url":"https://pubmed.ncbi.nlm.nih.gov/2275125","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.06.15.659737","title":"Cytoprotective roles of “E3 ubiquitin ligases-NF-κB-autophagy” axis in Pacific oysters  <i>Crassostrea gigas</i>  exposed to phenanthrene","date":"2025-06-18","source":"bioRxiv","url":"https://doi.org/10.1101/2025.06.15.659737","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13695,"output_tokens":3643,"usd":0.047865,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11316,"output_tokens":3961,"usd":0.077802,"stage2_stop_reason":"end_turn"},"total_usd":0.125667,"stage1_batch_id":"msgbatch_017qqVz33Ga63SCDwzb35XPb","stage2_batch_id":"msgbatch_01Scr2Ei9x4HJMiRGxwrmvLK","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"Haprin (TRIM36) is a haploid germ cell-specific RING finger / RBCC motif protein localized to the acrosomal region of elongated spermatids and mature sperm; a specific antibody against its RING finger domain inhibited the acrosome reaction in permeabilized sperm, establishing a direct role in acrosomal exocytosis.\",\n      \"method\": \"Western blot, immunohistochemistry, antibody inhibition of acrosome reaction in permeabilized sperm\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — localization by IHC linked to functional consequence via antibody inhibition assay, single lab with two orthogonal methods\",\n      \"pmids\": [\"12917430\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Human HAPRIN (TRIM36) is expressed exclusively in testes, localizes to the acrosomal region of sperm, and disappears after the acrosome reaction, indicating functional conservation of its role in the acrosome reaction between mouse and human.\",\n      \"method\": \"Western blot, immunocytochemistry in human testis and sperm\",\n      \"journal\": \"Journal of andrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization experiment with functional inference from disappearance post-reaction, single lab, two methods\",\n      \"pmids\": [\"15955891\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"TRIM36 has E3 ubiquitin ligase activity and interacts with the kinetochore protein CENP-H, co-localizes with alpha-tubulin (microtubules), and its overexpression decelerates cell cycle progression and attenuates cell growth, suggesting a role in chromosome segregation.\",\n      \"method\": \"Yeast two-hybrid screening (TRIM36–CENP-H interaction), immunofluorescence (colocalization with alpha-tubulin), cell growth/cycle assays upon overexpression\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — yeast two-hybrid for interaction, immunofluorescence for localization, overexpression phenotype; single lab, multiple methods but no in vitro reconstitution of ligase activity on CENP-H\",\n      \"pmids\": [\"19232519\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"In Xenopus, maternal trim36 mRNA is localized to the vegetal cortex/germ plasm and encodes a ubiquitin ligase required for cortical rotation and dorsoventral axis formation; depletion causes ventralized embryos rescued by wnt11 mRNA injection, placing Trim36 upstream of Wnt/beta-catenin activation. Ubiquitin ligase activity of Trim36 is required for vegetal microtubule polymerization and cortical rotation.\",\n      \"method\": \"Antisense oligonucleotide maternal loss-of-function, wnt11 mRNA rescue, egg tipping rescue, ubiquitin ligase-dead mutant analysis\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis by rescue experiment, ligase-dead mutant, multiple orthogonal rescue approaches, in a dedicated mechanistic study\",\n      \"pmids\": [\"19675128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Morpholino-mediated knockdown of Trim36/Haprin in Xenopus laevis specifically inhibits somite formation, establishing a role for this E3 ligase in somitogenesis during early embryogenesis.\",\n      \"method\": \"Morpholino antisense knockdown, temporal and spatial expression analysis (in situ hybridization, RT-PCR)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean morpholino knockdown with specific somite phenotype, single lab, single method for functional readout\",\n      \"pmids\": [\"19032936\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A homozygous missense mutation (p.Pro508Thr) in the B30.2/SPRY domain of TRIM36 causes autosomal recessive anencephaly; the mutant protein is less stable, and its transient expression in HeLa and LN229 cells disrupts microtubule organization, causes disorganized spindles, multiple spindles, abnormal cytokinesis, reduced proliferation, and increased apoptosis compared to wild-type TRIM36.\",\n      \"method\": \"Whole-exome sequencing, in silico conservation analysis, in vitro stability assay, transient transfection of mutant vs WT in HeLa/LN229 cells, immunofluorescence of microtubules/spindles, proliferation and apoptosis assays, siRNA knockdown\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (structural domain mutation, cell biology, knockdown), human disease linkage, replicated across two cell lines\",\n      \"pmids\": [\"28087737\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"TRIM36 inhibits prostate cancer cell cycle progression and cell proliferation in vitro and in vivo via inhibition of MAPK/ERK phosphorylation; restoring TRIM36 expression during anti-androgen therapy improves drug efficacy.\",\n      \"method\": \"Overexpression and knockdown in prostate cancer cell lines, cell cycle and proliferation assays, in vivo xenograft, phospho-ERK western blot\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean gain/loss-of-function with specific pathway readout (phospho-ERK), in vitro and in vivo, single lab\",\n      \"pmids\": [\"29449534\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Haprin (TRIM36)-deficient mice generated by homologous knockout show normal spermatogenesis but reduced sperm morphology and motility, and spermatozoa fail to fertilize oocytes under standard IVF conditions, demonstrating that Haprin is required for spermiogenesis and in vitro fertilization competence.\",\n      \"method\": \"Knockout mouse generation, spermatogenesis histology, sperm morphology and motility analysis, in vitro fertilization assay\",\n      \"journal\": \"Molecular reproduction and development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — germline KO mouse with multiple phenotypic readouts (morphology, motility, IVF), clean genetic model\",\n      \"pmids\": [\"32311190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TRIM36 promotes ubiquitination of beta-catenin, leading to its inactivation, thereby inhibiting Wnt/beta-catenin signaling and suppressing esophageal squamous cell carcinoma (ESCC) tumor growth in vitro and in vivo.\",\n      \"method\": \"Lentivirus-mediated overexpression/knockdown, ubiquitination assay of beta-catenin, flow cytometry (cell cycle, apoptosis), xenograft mouse model\",\n      \"journal\": \"Human cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ubiquitination assay combined with in vivo xenograft validation, single lab\",\n      \"pmids\": [\"35768649\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TRIM36 forms a complex with RAD51 and promotes its ubiquitination and degradation, thereby enhancing radiosensitivity in lung adenocarcinoma (LUAD) cells.\",\n      \"method\": \"Co-immunoprecipitation (TRIM36–RAD51 complex), ubiquitination assay, overexpression/knockdown with radiation treatment, proliferation and apoptosis assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP for complex, ubiquitination assay, functional radiation phenotype; single lab\",\n      \"pmids\": [\"36058131\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"TRIM36 is a direct target of miR-494-3p; TRIM36 promotes ubiquitination of cyclin E, reducing its levels and inhibiting HCC cell cycle progression and proliferation.\",\n      \"method\": \"Luciferase reporter assay (miR-494-3p targeting TRIM36 3'UTR), Co-IP and ubiquitination assay (TRIM36–cyclin E), overexpression/knockdown experiments, xenograft model\",\n      \"journal\": \"Journal of clinical and translational hepatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and ubiquitination assay for substrate identification, validated in vivo; single lab\",\n      \"pmids\": [\"36062277\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"TRIM36 directly interacts with FOXA2 and induces its K48-linked polyubiquitination, leading to FOXA2 protein degradation; loss of FOXA2 weakens NRF2 pathway activation and decreases GPX4 levels, inducing ferroptosis in colorectal cancer cells.\",\n      \"method\": \"Co-immunoprecipitation (TRIM36–FOXA2 interaction), ubiquitination assay (K48-linkage), overexpression/knockdown with NRF2/GPX4 western blot, ferroptosis assays\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus K48-linkage-specific ubiquitination assay with downstream pathway readout; single lab\",\n      \"pmids\": [\"37875418\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM36 mediates K48-linked polyubiquitination of phospho-AKT1 (at T308/S473), leading to its proteasomal degradation and inhibition of AKT signaling, thereby suppressing lung fibroblast activation and pulmonary fibrosis; TRIM36 overexpression ameliorates bleomycin-induced pulmonary fibrosis in mice.\",\n      \"method\": \"Co-immunoprecipitation, K48-linked ubiquitination assay (phospho-AKT1 substrate), overexpression in fibroblasts, bleomycin mouse model\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assay, in vivo validation, single lab\",\n      \"pmids\": [\"41323265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM36 directly binds GRB7 and promotes its ubiquitination and degradation; TRIM36 knockout in mice promotes AOM/DSS-induced colorectal carcinogenesis and inhibits autophagy, and this tumor-suppressive role is mediated through GRB7 degradation.\",\n      \"method\": \"Co-immunoprecipitation (TRIM36–GRB7), ubiquitination assay, TRIM36 knockout mice (AOM/DSS model), overexpression/knockdown in CRC cell lines\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and ubiquitination assay plus in vivo KO model; single lab\",\n      \"pmids\": [\"39803720\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"TRIM36 promotes K48-linked polyubiquitination and proteasomal degradation of DDX3X, shifting macrophage polarization from M1 to M2 phenotype and ameliorating tubal factor infertility in a rat model.\",\n      \"method\": \"Co-immunoprecipitation (TRIM36–DDX3X), ubiquitination assay (K48-linkage), TRIM36 overexpression in BMDMs, in vivo rat TFI model with oe-TRIM36 therapy\",\n      \"journal\": \"Cell biology and toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP and linkage-specific ubiquitination assay, in vivo validation in rat model, single lab\",\n      \"pmids\": [\"41553545\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TRIM36 is a microtubule-associated RING-finger/RBCC E3 ubiquitin ligase that promotes K48-linked polyubiquitination and proteasomal degradation of multiple substrates—including FOXA2, beta-catenin, RAD51, cyclin E, phospho-AKT1, GRB7, and DDX3X—thereby regulating cell cycle progression, ferroptosis, Wnt/beta-catenin and MAPK/ERK signaling, and cytoskeletal organization; in the germline it localizes to the sperm acrosome and is required for the acrosome reaction and fertilization, while in early vertebrate development it controls microtubule polymerization, cortical rotation, and dorsoventral axis specification upstream of Wnt/beta-catenin.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TRIM36 is a microtubule-associated RING-finger/RBCC-family E3 ubiquitin ligase that controls cytoskeletal organization, cell cycle progression, and developmental patterning, and that acts as a substrate-selective degradation factor across multiple signaling pathways [#2, #3]. It possesses E3 ligase activity, colocalizes with alpha-tubulin and the kinetochore protein CENP-H, and its overexpression slows cell cycle progression, consistent with a role in chromosome segregation and microtubule-dependent processes [#2]. In early Xenopus development its ligase activity drives vegetal microtubule polymerization and cortical rotation required for dorsoventral axis formation, acting upstream of Wnt/beta-catenin signaling, and it is also required for somitogenesis [#3, #4]. As a degradative ligase, TRIM36 directs K48-linked polyubiquitination and proteasomal turnover of a series of substrates — FOXA2 (linking it to NRF2/GPX4-dependent ferroptosis), phospho-AKT1, DDX3X, GRB7, RAD51, and cyclin E — and promotes ubiquitination of beta-catenin, thereby suppressing Wnt/beta-catenin and MAPK/ERK signaling and acting as a tumor suppressor in prostate, esophageal, hepatocellular, lung, and colorectal cancers [#6, #8, #9, #10, #11, #12, #13, #14]. In the germline, TRIM36 (Haprin) localizes to the sperm acrosomal region and is required for the acrosome reaction and fertilization competence: antibody inhibition blocks acrosomal exocytosis and knockout mice produce morphologically abnormal, poorly motile sperm that fail in IVF [#0, #1, #7]. A homozygous B30.2/SPRY-domain missense mutation (p.Pro508Thr) causes autosomal recessive anencephaly, destabilizing the protein and disrupting microtubule and spindle organization [#5].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established TRIM36/Haprin as a germ-cell RBCC protein with a direct functional role in sperm, answering whether this RING-finger protein participates in acrosomal exocytosis.\",\n      \"evidence\": \"Western blot and IHC localization to the acrosomal region plus antibody inhibition of the acrosome reaction in permeabilized mouse sperm\",\n      \"pmids\": [\"12917430\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not identify ubiquitination substrates in sperm\", \"Mechanism linking the RING domain to exocytosis not defined\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Showed the sperm acrosomal role is conserved in human, testing whether mouse findings generalize across species.\",\n      \"evidence\": \"Western blot and immunocytochemistry in human testis and sperm, with loss of signal after the acrosome reaction\",\n      \"pmids\": [\"15955891\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional perturbation in human sperm not performed\", \"No substrate identified\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined TRIM36 as an active E3 ligase associated with microtubules and the kinetochore, addressing its molecular activity and cell-division role beyond the germline.\",\n      \"evidence\": \"Yeast two-hybrid identifying CENP-H interaction, immunofluorescence colocalization with alpha-tubulin, and overexpression cell cycle/growth assays\",\n      \"pmids\": [\"19232519\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution of ligase activity on CENP-H\", \"CENP-H not shown to be a degradation substrate\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Implicated Trim36 in somitogenesis, extending its developmental requirement beyond axis formation.\",\n      \"evidence\": \"Morpholino knockdown in Xenopus laevis with in situ/RT-PCR expression analysis\",\n      \"pmids\": [\"19032936\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Pathway connecting TRIM36 to somite formation undefined\", \"Single functional readout\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Placed Trim36 ligase activity upstream of Wnt/beta-catenin in vertebrate dorsoventral patterning by showing it drives the cytoskeletal events of cortical rotation, the strongest mechanistic anchor for its developmental function.\",\n      \"evidence\": \"Maternal antisense loss-of-function, wnt11 mRNA and egg-tipping rescue, and ligase-dead mutant analysis in Xenopus\",\n      \"pmids\": [\"19675128\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The ubiquitination substrate linking ligase activity to microtubule polymerization not identified\", \"Direct biochemical target in cortical rotation unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Linked TRIM36 to human disease and to spindle/microtubule integrity, establishing a Mendelian phenotype and a cell-division function for the protein.\",\n      \"evidence\": \"Whole-exome sequencing identifying p.Pro508Thr in the B30.2/SPRY domain, stability assays, and transfection of mutant vs WT in HeLa/LN229 with spindle, proliferation and apoptosis readouts\",\n      \"pmids\": [\"28087737\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the SPRY-domain mutation impairs substrate recognition not resolved\", \"Causal substrate underlying spindle defects unidentified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified TRIM36 as a tumor suppressor acting through MAPK/ERK, beginning the cancer-pathway phase of its characterization.\",\n      \"evidence\": \"Gain/loss-of-function in prostate cancer lines, phospho-ERK western blot, proliferation/cell cycle assays, and xenografts\",\n      \"pmids\": [\"29449534\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitination substrate in the ERK axis not defined\", \"Mechanism of ERK suppression unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Established TRIM36 as a substrate-selective K48-ligase across several oncogenic effectors, defining its degradative mechanism for beta-catenin, RAD51, and cyclin E.\",\n      \"evidence\": \"Co-IP and ubiquitination assays for beta-catenin (ESCC), RAD51 (LUAD radiosensitivity), and cyclin E (HCC, downstream of miR-494-3p), with overexpression/knockdown and xenografts\",\n      \"pmids\": [\"35768649\", \"36058131\", \"36062277\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ubiquitination linkage type not specified for all substrates\", \"Substrate recognition determinants not mapped\", \"Single-lab reports per substrate\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended the K48-degradation mechanism to FOXA2 and connected it to ferroptosis, showing TRIM36 controls the NRF2/GPX4 antioxidant axis.\",\n      \"evidence\": \"Co-IP, K48-linkage-specific ubiquitination assay, and NRF2/GPX4 western blot with ferroptosis assays in colorectal cancer cells\",\n      \"pmids\": [\"37875418\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct enzymatic reconstitution absent\", \"In vivo contribution to ferroptosis not tested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Broadened the substrate repertoire to phospho-AKT1 and GRB7, linking TRIM36 to AKT signaling in fibrosis and to autophagy-dependent tumor suppression in vivo.\",\n      \"evidence\": \"Co-IP and K48-ubiquitination assays for phospho-AKT1 (bleomycin pulmonary fibrosis) and GRB7 (AOM/DSS colorectal model in TRIM36-KO mice)\",\n      \"pmids\": [\"41323265\", \"39803720\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Selectivity for phospho-AKT1 over unphosphorylated AKT1 not mechanistically explained\", \"Each substrate validated by single lab\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Connected TRIM36-mediated DDX3X degradation to macrophage polarization, extending its degradative role into immune phenotype control.\",\n      \"evidence\": \"Co-IP, K48-linkage ubiquitination assay, overexpression in BMDMs, and an in vivo rat tubal factor infertility model\",\n      \"pmids\": [\"41553545\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism linking DDX3X loss to M1-to-M2 switch not fully defined\", \"Single-model validation\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A unifying biochemical picture of how TRIM36 selects its diverse substrates and how its microtubule/developmental functions relate to its degradative oncosuppressive functions remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of substrate recognition by the B30.2/SPRY domain\", \"No reconstituted in vitro ligase assay defining minimal E2/substrate requirements\", \"Whether the developmental microtubule role and the cancer substrate-degradation roles share a common molecular mechanism is untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [2, 3, 8, 11, 12]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [2, 3, 9, 10, 11, 13, 14]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [8, 11, 12, 13, 14]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [2, 3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [2, 3, 5]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [6, 8, 12]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [3, 4]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [8, 11, 12, 13, 14]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2, 10]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [0, 7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"CENP-H\", \"FOXA2\", \"RAD51\", \"GRB7\", \"DDX3X\", \"CTNNB1\", \"AKT1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}