{"gene":"TCP10L","run_date":"2026-06-10T10:51:54","timeline":{"discoveries":[{"year":2003,"finding":"TCP10L encodes a leucine zipper-containing protein that localizes to the nucleus and functions as a transcriptional repressor; luciferase reporter assays showed ~2.6–9.8 fold repression of reporter gene expression in HEK293, SK-HEP-1, and CHO cells, and a leucine zipper mutant almost completely abolished this repression, indicating the leucine zipper is critical for transcriptional inhibition activity.","method":"Dual luciferase reporter assay with wild-type and leucine zipper mutant TCP10L; TCP10L-EGFP subcellular localization by fluorescence imaging","journal":"Journal of human genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional reporter assay with mutagenesis and localization, single lab, two orthogonal methods","pmids":["14586771"],"is_preprint":false},{"year":2004,"finding":"TCP10L physically interacts with MAD4 (a MYC-antagonizing transcription factor); interaction identified by yeast two-hybrid and confirmed by co-immunoprecipitation and co-localization experiments.","method":"Yeast two-hybrid screen, co-immunoprecipitation, subcellular localization","journal":"Journal of biochemistry and molecular biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal confirmation by Co-IP after Y2H, single lab, two orthogonal methods","pmids":["15469726"],"is_preprint":false},{"year":2005,"finding":"TCP10L is expressed specifically in the nucleus of spermatogenic cells during spermatogenesis and binds to death-associated protein kinase 3 (DAPK-3/ZIP kinase); the interaction depends on the leucine zipper motif-containing region of TCP10L, as shown by mutagenesis.","method":"Yeast two-hybrid screening, co-immunoprecipitation, subcellular localization, mutagenesis","journal":"International journal of andrology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Y2H confirmed by Co-IP and localization, mutagenesis defining domain, single lab","pmids":["15910542"],"is_preprint":false},{"year":2007,"finding":"TCP10L homodimerizes through its leucine zipper motif; deletion or point mutation of the leucine zipper abolishes homodimerization in vitro and in vivo, but leucine zipper mutants still localize to the nucleus, indicating the leucine zipper is required for dimerization but not for nuclear localization.","method":"In vitro and in vivo homodimerization assays, leucine zipper deletion and point mutation analysis, immunofluorescence in HeLa cells","journal":"Molecular biology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro and in vivo methods with mutagenesis, single lab","pmids":["17377852"],"is_preprint":false},{"year":2014,"finding":"TCP10L acts as a tumor suppressor in hepatocellular carcinoma by inhibiting cell cycle progression at G0/G1 phase; overexpression suppressed colony formation and attenuated cell growth in vivo, while silencing promoted cell cycle progression and cell growth.","method":"TCP10L overexpression and siRNA knockdown in HCC cells; colony formation assay, cell cycle analysis, in vivo tumor growth assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — bidirectional (OE and KD) loss/gain of function with defined cell cycle phenotype, single lab","pmids":["24565846"],"is_preprint":false},{"year":2016,"finding":"TCP10L interacts with MAD1 through their respective leucine zipper domains; TCP10L stabilizes intracellular MAD1 protein levels through this direct interaction, synergizes with MAD1 in transcriptional repression and G1 cell cycle arrest, and the interaction-deficient TCP10L mutant fails to stabilize MAD1 or suppress cell growth.","method":"Co-immunoprecipitation, leucine zipper domain mutants, luciferase transcriptional repression assay, cell cycle analysis, Western blot for MAD1 protein levels","journal":"BMB reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with domain mutants plus multiple functional readouts, single lab, orthogonal methods","pmids":["26698869"],"is_preprint":false},{"year":2025,"finding":"NEK6, a serine/threonine kinase, binds to TCP10L and degrades TCP10L protein through ubiquitination, thereby negatively regulating TCP10L expression; rescue experiments demonstrated that TCP10L reverses the pro-tumorigenic and pro-glycolytic effects of NEK6 in HCC cells.","method":"Co-immunoprecipitation (NEK6–TCP10L binding), ubiquitination assay, NEK6 knockdown with TCP10L rescue experiments in HCC cells","journal":"Critical reviews in eukaryotic gene expression","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus ubiquitination assay plus rescue experiments, single lab","pmids":["40228222"],"is_preprint":false}],"current_model":"TCP10L is a primate-specific, liver- and testis-expressed nuclear protein that homodimerizes via its leucine zipper motif and functions as a transcriptional repressor; it physically interacts with MAD1 (stabilizing it), MAD4, and DAPK-3 through its leucine zipper domain, synergizes with MAD1 to enforce G1 cell cycle arrest and suppress cell growth, and its protein level is regulated by NEK6-mediated ubiquitination and degradation."},"narrative":{"mechanistic_narrative":"TCP10L is a nuclear, leucine zipper-containing transcriptional repressor expressed in liver and in the nucleus of spermatogenic cells, where it restrains cell growth and cell cycle progression [PMID:14586771, PMID:15910542, PMID:24565846]. Its leucine zipper motif mediates homodimerization and is essential both for transcriptional repression activity and for protein-protein interactions, while nuclear localization is independent of this motif [PMID:14586771, PMID:17377852]. Through its leucine zipper domain TCP10L engages the MYC-antagonizing transcription factors MAD4 and MAD1 as well as DAPK-3/ZIP kinase; binding to MAD1 stabilizes MAD1 protein levels and synergizes with it to drive transcriptional repression and G1 cell cycle arrest, an activity lost in interaction-deficient mutants [PMID:15469726, PMID:15910542, PMID:26698869]. Consistent with this growth-suppressive role, TCP10L acts as a tumor suppressor in hepatocellular carcinoma, arresting cells at G0/G1 and suppressing colony formation and tumor growth, and its level is negatively controlled by NEK6, which binds and targets TCP10L for ubiquitination and degradation to relieve its suppression of tumorigenic and glycolytic programs [PMID:24565846, PMID:40228222].","teleology":[{"year":2003,"claim":"Established that TCP10L is a nuclear protein with intrinsic transcriptional repressor activity dependent on its leucine zipper, defining its core molecular function.","evidence":"Dual luciferase reporter assays with wild-type and leucine zipper mutant plus EGFP localization in HEK293, SK-HEP-1, and CHO cells","pmids":["14586771"],"confidence":"Medium","gaps":["No direct DNA-binding target or promoter identified","Repression mechanism (co-repressor recruitment) not defined"]},{"year":2004,"claim":"Identified the first interaction partner, MAD4, linking TCP10L to the MYC/MAD transcriptional network.","evidence":"Yeast two-hybrid screen confirmed by co-immunoprecipitation and co-localization","pmids":["15469726"],"confidence":"Medium","gaps":["Functional consequence of the MAD4 interaction not established","Interaction domain not mapped in this study"]},{"year":2005,"claim":"Showed TCP10L is expressed in spermatogenic cell nuclei and binds DAPK-3/ZIP kinase via its leucine zipper region, extending its partner repertoire to a kinase.","evidence":"Yeast two-hybrid, co-immunoprecipitation, localization, and mutagenesis","pmids":["15910542"],"confidence":"Medium","gaps":["Functional outcome of DAPK-3 binding (phosphorylation of TCP10L?) not determined","Role in spermatogenesis not functionally tested"]},{"year":2007,"claim":"Resolved that the leucine zipper drives homodimerization but is dispensable for nuclear import, separating the dimerization function from localization.","evidence":"In vitro and in vivo homodimerization assays with leucine zipper deletion/point mutants and immunofluorescence in HeLa cells","pmids":["17377852"],"confidence":"Medium","gaps":["Nuclear localization signal not mapped","Whether homodimers vs heterodimers dictate repressor activity unresolved"]},{"year":2014,"claim":"Demonstrated a physiological growth-suppressive role: TCP10L is a hepatocellular carcinoma tumor suppressor that enforces G0/G1 arrest.","evidence":"Overexpression and siRNA knockdown in HCC cells with colony formation, cell cycle analysis, and in vivo tumor growth assays","pmids":["24565846"],"confidence":"Medium","gaps":["Direct downstream cell cycle effectors not identified","Link between transcriptional repression and arrest mechanistically unspecified"]},{"year":2016,"claim":"Provided the mechanistic basis for growth suppression: TCP10L binds and stabilizes MAD1 via leucine zipper domains and synergizes with it for repression and G1 arrest.","evidence":"Co-immunoprecipitation with domain mutants, luciferase repression assays, cell cycle analysis, and MAD1 Western blots","pmids":["26698869"],"confidence":"Medium","gaps":["Mechanism by which TCP10L stabilizes MAD1 (blocking degradation?) not defined","Shared target genes of TCP10L/MAD1 not identified"]},{"year":2025,"claim":"Identified upstream regulation of TCP10L: NEK6 binds and ubiquitinates TCP10L for degradation, and restoring TCP10L reverses NEK6-driven tumorigenesis and glycolysis.","evidence":"Co-immunoprecipitation, ubiquitination assay, and NEK6 knockdown with TCP10L rescue in HCC cells","pmids":["40228222"],"confidence":"Medium","gaps":["Whether NEK6 directly ubiquitinates or recruits an E3 ligase not resolved","Ubiquitination sites on TCP10L not mapped"]},{"year":null,"claim":"The direct DNA targets and the full transcriptional program controlled by TCP10L, and its in vivo function in spermatogenesis, remain undefined.","evidence":"No genome-wide target or knockout phenotype reported in the available corpus","pmids":[],"confidence":"Medium","gaps":["No ChIP/genome-wide target data","No organismal loss-of-function model","Mechanistic basis of testis-specific role unaddressed"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[0,5]},{"term_id":"GO:0042393","term_label":"histone binding","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,2,3]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[4,5]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[0]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,6]}],"complexes":[],"partners":["MAD1","MAD4","DAPK3","NEK6"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8TDR4","full_name":"T-complex protein 10A homolog 1","aliases":["TCP10-like"],"length_aa":215,"mass_kda":23.9,"function":"May be involved in transcriptional regulation. Has in vitro transcription inhibition activity. Acts as a tumor suppressor in hepatocellular carcinoma (HCC) cells","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q8TDR4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/TCP10L","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/TCP10L","total_profiled":1310},"omim":[{"mim_id":"620016","title":"MAX DIMERIZATION PROTEIN 4; MXD4","url":"https://www.omim.org/entry/620016"},{"mim_id":"608365","title":"T-COMPLEX PROTEIN 10-LIKE; TCP10L","url":"https://www.omim.org/entry/608365"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enriched","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"testis","ntpm":142.6}],"url":"https://www.proteinatlas.org/search/TCP10L"},"hgnc":{"alias_symbol":["PRED77"],"prev_symbol":[]},"alphafold":{"accession":"Q8TDR4","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8TDR4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8TDR4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8TDR4-F1-predicted_aligned_error_v6.png","plddt_mean":66.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=TCP10L","jax_strain_url":"https://www.jax.org/strain/search?query=TCP10L"},"sequence":{"accession":"Q8TDR4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8TDR4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8TDR4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8TDR4"}},"corpus_meta":[{"pmid":"28803425","id":"PMC_28803425","title":"Frameshift Mutations in Repeat Sequences of ANK3, HACD4, TCP10L, TP53BP1, MFN1, LCMT2, RNMT, TRMT6, METTL8 and METTL16 Genes in Colon Cancers.","date":"2017","source":"Pathology oncology research : POR","url":"https://pubmed.ncbi.nlm.nih.gov/28803425","citation_count":47,"is_preprint":false},{"pmid":"15910542","id":"PMC_15910542","title":"TCP10L is expressed specifically in spermatogenic cells and binds to death associated protein kinase-3.","date":"2005","source":"International journal of andrology","url":"https://pubmed.ncbi.nlm.nih.gov/15910542","citation_count":12,"is_preprint":false},{"pmid":"17377852","id":"PMC_17377852","title":"Identification of TCP10L as primate-specific gene derived via segmental duplication and homodimerization of TCP10L through the leucine zipper motif.","date":"2007","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/17377852","citation_count":6,"is_preprint":false},{"pmid":"24565846","id":"PMC_24565846","title":"TCP10L acts as a tumor suppressor by inhibiting cell proliferation in hepatocellular carcinoma.","date":"2014","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/24565846","citation_count":5,"is_preprint":false},{"pmid":"14586771","id":"PMC_14586771","title":"Identification of a novel liver-specific expressed gene, TCP10L, encoding a human leucine zipper protein with transcription inhibition activity.","date":"2003","source":"Journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/14586771","citation_count":5,"is_preprint":false},{"pmid":"15469726","id":"PMC_15469726","title":"Human liver specific transcriptional factor TCP10L binds to MAD4.","date":"2004","source":"Journal of biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/15469726","citation_count":5,"is_preprint":false},{"pmid":"31671693","id":"PMC_31671693","title":"Novel Mutations Segregating with Complete Androgen Insensitivity Syndrome and their Molecular Characteristics.","date":"2019","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/31671693","citation_count":5,"is_preprint":false},{"pmid":"40228222","id":"PMC_40228222","title":"NEK6 Accelerates Hepatocellular Carcinoma Progression and Glycolysis through Ubiquitination of TCP10L.","date":"2025","source":"Critical reviews in eukaryotic gene expression","url":"https://pubmed.ncbi.nlm.nih.gov/40228222","citation_count":3,"is_preprint":false},{"pmid":"33947493","id":"PMC_33947493","title":"Whole-Genome Study of a Multigenerational Family with Essential Tremor.","date":"2021","source":"The Canadian journal of neurological sciences. Le journal canadien des sciences neurologiques","url":"https://pubmed.ncbi.nlm.nih.gov/33947493","citation_count":3,"is_preprint":false},{"pmid":"26698869","id":"PMC_26698869","title":"TCP10L synergizes with MAD1 in transcriptional suppression and cell cycle arrest through mutual interaction.","date":"2016","source":"BMB reports","url":"https://pubmed.ncbi.nlm.nih.gov/26698869","citation_count":2,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6527,"output_tokens":1657,"usd":0.022218,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8511,"output_tokens":2464,"usd":0.052077,"stage2_stop_reason":"end_turn"},"total_usd":0.074295,"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\": \"TCP10L encodes a leucine zipper-containing protein that localizes to the nucleus and functions as a transcriptional repressor; luciferase reporter assays showed ~2.6–9.8 fold repression of reporter gene expression in HEK293, SK-HEP-1, and CHO cells, and a leucine zipper mutant almost completely abolished this repression, indicating the leucine zipper is critical for transcriptional inhibition activity.\",\n      \"method\": \"Dual luciferase reporter assay with wild-type and leucine zipper mutant TCP10L; TCP10L-EGFP subcellular localization by fluorescence imaging\",\n      \"journal\": \"Journal of human genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional reporter assay with mutagenesis and localization, single lab, two orthogonal methods\",\n      \"pmids\": [\"14586771\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"TCP10L physically interacts with MAD4 (a MYC-antagonizing transcription factor); interaction identified by yeast two-hybrid and confirmed by co-immunoprecipitation and co-localization experiments.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, subcellular localization\",\n      \"journal\": \"Journal of biochemistry and molecular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal confirmation by Co-IP after Y2H, single lab, two orthogonal methods\",\n      \"pmids\": [\"15469726\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"TCP10L is expressed specifically in the nucleus of spermatogenic cells during spermatogenesis and binds to death-associated protein kinase 3 (DAPK-3/ZIP kinase); the interaction depends on the leucine zipper motif-containing region of TCP10L, as shown by mutagenesis.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation, subcellular localization, mutagenesis\",\n      \"journal\": \"International journal of andrology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Y2H confirmed by Co-IP and localization, mutagenesis defining domain, single lab\",\n      \"pmids\": [\"15910542\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"TCP10L homodimerizes through its leucine zipper motif; deletion or point mutation of the leucine zipper abolishes homodimerization in vitro and in vivo, but leucine zipper mutants still localize to the nucleus, indicating the leucine zipper is required for dimerization but not for nuclear localization.\",\n      \"method\": \"In vitro and in vivo homodimerization assays, leucine zipper deletion and point mutation analysis, immunofluorescence in HeLa cells\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro and in vivo methods with mutagenesis, single lab\",\n      \"pmids\": [\"17377852\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"TCP10L acts as a tumor suppressor in hepatocellular carcinoma by inhibiting cell cycle progression at G0/G1 phase; overexpression suppressed colony formation and attenuated cell growth in vivo, while silencing promoted cell cycle progression and cell growth.\",\n      \"method\": \"TCP10L overexpression and siRNA knockdown in HCC cells; colony formation assay, cell cycle analysis, in vivo tumor growth assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — bidirectional (OE and KD) loss/gain of function with defined cell cycle phenotype, single lab\",\n      \"pmids\": [\"24565846\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"TCP10L interacts with MAD1 through their respective leucine zipper domains; TCP10L stabilizes intracellular MAD1 protein levels through this direct interaction, synergizes with MAD1 in transcriptional repression and G1 cell cycle arrest, and the interaction-deficient TCP10L mutant fails to stabilize MAD1 or suppress cell growth.\",\n      \"method\": \"Co-immunoprecipitation, leucine zipper domain mutants, luciferase transcriptional repression assay, cell cycle analysis, Western blot for MAD1 protein levels\",\n      \"journal\": \"BMB reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with domain mutants plus multiple functional readouts, single lab, orthogonal methods\",\n      \"pmids\": [\"26698869\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NEK6, a serine/threonine kinase, binds to TCP10L and degrades TCP10L protein through ubiquitination, thereby negatively regulating TCP10L expression; rescue experiments demonstrated that TCP10L reverses the pro-tumorigenic and pro-glycolytic effects of NEK6 in HCC cells.\",\n      \"method\": \"Co-immunoprecipitation (NEK6–TCP10L binding), ubiquitination assay, NEK6 knockdown with TCP10L rescue experiments in HCC cells\",\n      \"journal\": \"Critical reviews in eukaryotic gene expression\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus ubiquitination assay plus rescue experiments, single lab\",\n      \"pmids\": [\"40228222\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"TCP10L is a primate-specific, liver- and testis-expressed nuclear protein that homodimerizes via its leucine zipper motif and functions as a transcriptional repressor; it physically interacts with MAD1 (stabilizing it), MAD4, and DAPK-3 through its leucine zipper domain, synergizes with MAD1 to enforce G1 cell cycle arrest and suppress cell growth, and its protein level is regulated by NEK6-mediated ubiquitination and degradation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"TCP10L is a nuclear, leucine zipper-containing transcriptional repressor expressed in liver and in the nucleus of spermatogenic cells, where it restrains cell growth and cell cycle progression [#0, #2, #4]. Its leucine zipper motif mediates homodimerization and is essential both for transcriptional repression activity and for protein-protein interactions, while nuclear localization is independent of this motif [#0, #3]. Through its leucine zipper domain TCP10L engages the MYC-antagonizing transcription factors MAD4 and MAD1 as well as DAPK-3/ZIP kinase; binding to MAD1 stabilizes MAD1 protein levels and synergizes with it to drive transcriptional repression and G1 cell cycle arrest, an activity lost in interaction-deficient mutants [#1, #2, #5]. Consistent with this growth-suppressive role, TCP10L acts as a tumor suppressor in hepatocellular carcinoma, arresting cells at G0/G1 and suppressing colony formation and tumor growth, and its level is negatively controlled by NEK6, which binds and targets TCP10L for ubiquitination and degradation to relieve its suppression of tumorigenic and glycolytic programs [#4, #6].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established that TCP10L is a nuclear protein with intrinsic transcriptional repressor activity dependent on its leucine zipper, defining its core molecular function.\",\n      \"evidence\": \"Dual luciferase reporter assays with wild-type and leucine zipper mutant plus EGFP localization in HEK293, SK-HEP-1, and CHO cells\",\n      \"pmids\": [\"14586771\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct DNA-binding target or promoter identified\", \"Repression mechanism (co-repressor recruitment) not defined\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Identified the first interaction partner, MAD4, linking TCP10L to the MYC/MAD transcriptional network.\",\n      \"evidence\": \"Yeast two-hybrid screen confirmed by co-immunoprecipitation and co-localization\",\n      \"pmids\": [\"15469726\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the MAD4 interaction not established\", \"Interaction domain not mapped in this study\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Showed TCP10L is expressed in spermatogenic cell nuclei and binds DAPK-3/ZIP kinase via its leucine zipper region, extending its partner repertoire to a kinase.\",\n      \"evidence\": \"Yeast two-hybrid, co-immunoprecipitation, localization, and mutagenesis\",\n      \"pmids\": [\"15910542\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional outcome of DAPK-3 binding (phosphorylation of TCP10L?) not determined\", \"Role in spermatogenesis not functionally tested\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Resolved that the leucine zipper drives homodimerization but is dispensable for nuclear import, separating the dimerization function from localization.\",\n      \"evidence\": \"In vitro and in vivo homodimerization assays with leucine zipper deletion/point mutants and immunofluorescence in HeLa cells\",\n      \"pmids\": [\"17377852\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Nuclear localization signal not mapped\", \"Whether homodimers vs heterodimers dictate repressor activity unresolved\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated a physiological growth-suppressive role: TCP10L is a hepatocellular carcinoma tumor suppressor that enforces G0/G1 arrest.\",\n      \"evidence\": \"Overexpression and siRNA knockdown in HCC cells with colony formation, cell cycle analysis, and in vivo tumor growth assays\",\n      \"pmids\": [\"24565846\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct downstream cell cycle effectors not identified\", \"Link between transcriptional repression and arrest mechanistically unspecified\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Provided the mechanistic basis for growth suppression: TCP10L binds and stabilizes MAD1 via leucine zipper domains and synergizes with it for repression and G1 arrest.\",\n      \"evidence\": \"Co-immunoprecipitation with domain mutants, luciferase repression assays, cell cycle analysis, and MAD1 Western blots\",\n      \"pmids\": [\"26698869\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which TCP10L stabilizes MAD1 (blocking degradation?) not defined\", \"Shared target genes of TCP10L/MAD1 not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified upstream regulation of TCP10L: NEK6 binds and ubiquitinates TCP10L for degradation, and restoring TCP10L reverses NEK6-driven tumorigenesis and glycolysis.\",\n      \"evidence\": \"Co-immunoprecipitation, ubiquitination assay, and NEK6 knockdown with TCP10L rescue in HCC cells\",\n      \"pmids\": [\"40228222\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether NEK6 directly ubiquitinates or recruits an E3 ligase not resolved\", \"Ubiquitination sites on TCP10L not mapped\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The direct DNA targets and the full transcriptional program controlled by TCP10L, and its in vivo function in spermatogenesis, remain undefined.\",\n      \"evidence\": \"No genome-wide target or knockout phenotype reported in the available corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No ChIP/genome-wide target data\", \"No organismal loss-of-function model\", \"Mechanistic basis of testis-specific role unaddressed\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [0, 5]},\n      {\"term_id\": \"GO:0042393\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 2, 3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [4, 5]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"MAD1\", \"MAD4\", \"DAPK3\", \"NEK6\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}