{"gene":"THUMPD2","run_date":"2026-06-10T10:51:55","timeline":{"discoveries":[{"year":2023,"finding":"THUMPD2 is an active N2-methylguanosine (m2G) methyltransferase that directly associates with U6 snRNA and is required for formation of the m2G modification on U6 snRNA (the last 'orphan' modification in U6 snRNA). THUMPD2 interacts with the co-factor TRMT112, which is required for its metabolic stability and enzymatic activity.","method":"Interaction network mapping via SILAC/pulldown in intact cells; in vitro methyltransferase activity assays; direct association with U6 snRNA demonstrated experimentally","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — enzymatic activity established in vitro, replicated independently in two separate papers (PMID:37283053 and PMID:38165050) using orthogonal methods","pmids":["37283053"],"is_preprint":false},{"year":2024,"finding":"THUMPD2 is the methyltransferase responsible for N2-methylation of G72 of U6 snRNA (m2G72) by explicitly recognizing U6-specific sequences and structural elements. Knockout of THUMPD2 eliminates U6 m2G72 and impairs pre-mRNA splicing activity, causing thousands of altered alternative splicing events in human cells and activating the nonsense-mediated mRNA decay pathway. The m2G modification at G72 of U6 snRNA is conserved among vertebrates.","method":"THUMPD2 knockout in human cells; in vitro methyltransferase assays with mutagenesis of U6 recognition elements; transcriptome-wide alternative splicing analysis; NMD pathway activation assayed","journal":"Nucleic acids research","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — enzymatic mechanism established by knockout + in vitro reconstitution with specificity mutagenesis, replicated across two independent studies","pmids":["38165050"],"is_preprint":false},{"year":2021,"finding":"THUMPD2 was identified as one of seven methyltransferase interaction partners of TRMT112 in a SILAC pulldown screen. TRMT112 stabilizes THUMPD2 in cells, and TRMT112 and THUMPD2 exhibit a strong mutual feedback loop when co-expressed.","method":"SILAC-based affinity pulldown; co-expression stability assays; single amino acid mutations on TRMT112 surface to probe interaction differences","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — SILAC pulldown with co-expression validation, single lab, consistent with independent replication in PMID:37283053","pmids":["34948388"],"is_preprint":false},{"year":2001,"finding":"THUMPD2 (C2orf8) encodes a putative 244 amino acid protein containing three methyltransferase motifs conserved across prokaryotic and eukaryotic species, and is expressed in multiple tissues. The gene was localized to chromosome 2p22→p21 by FISH. Alternative transcripts share a coding region spanning exons 6–10.","method":"RACE analysis, RT-PCR, Northern blot, FISH, computational ORF and domain analysis","journal":"Cytogenetics and cell genetics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — genomic localization and domain identification by multiple methods, but functional enzymatic activity not experimentally confirmed in this study","pmids":["12063391"],"is_preprint":false},{"year":2019,"finding":"Downregulation of THUMPD2 expression (via transposon insertion) and siRNA-mediated knockdown of THUMPD2 conferred resistance to both cisplatin and 5-fluorouracil in human esophageal squamous cell carcinoma cells in vitro.","method":"Transposon activation mutagenesis screen; siRNA knockdown; IC50 comparison between drug-resistant and wild-type cells","journal":"The journal of gene medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single knockdown approach with limited mechanistic follow-up; no pathway placement for the drug resistance phenotype","pmids":["31656051"],"is_preprint":false},{"year":2024,"finding":"shRNA-mediated knockdown of THUMPD2 in ovarian cancer cell lines (OVCAR3, SKOV3) increased cell proliferation but inhibited metastasis, while THUMPD2 overexpression had the opposite effect, with THUMPD2 overexpression suppressing tumor growth in vivo. GO/KEGG pathway analysis of THUMPD2 target genes implicated centrosome, microtubule, cell cycle, and extracellular matrix pathways.","method":"shRNA knockdown; overexpression; in vivo tumor growth assay; GO/KEGG analysis of transcriptome data","journal":"Heliyon","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, phenotypic readout without direct molecular mechanism established for THUMPD2's role in proliferation/metastasis","pmids":["39071668"],"is_preprint":false}],"current_model":"THUMPD2 is a TRMT112-dependent N2-methylguanosine (m2G) methyltransferase that specifically recognizes U6 snRNA sequence and structural elements to install the m2G modification at position G72 of U6 snRNA (the catalytic center of the spliceosome), thereby promoting efficient pre-mRNA splicing; loss of THUMPD2 eliminates this modification and globally disrupts alternative splicing, triggering nonsense-mediated mRNA decay."},"narrative":{"mechanistic_narrative":"THUMPD2 is an RNA methyltransferase that installs the N2-methylguanosine (m2G) modification on U6 small nuclear RNA, thereby promoting spliceosome function and accurate pre-mRNA splicing [PMID:37283053, PMID:38165050]. It directly associates with U6 snRNA and catalyzes m2G specifically at position G72, recognizing U6-specific sequence and structural elements, and its activity and metabolic stability depend on the co-factor TRMT112 [PMID:37283053, PMID:38165050]. Loss of THUMPD2 eliminates U6 m2G72, alters thousands of alternative splicing events, and activates the nonsense-mediated mRNA decay pathway, establishing this single modification as functionally important for splicing fidelity [PMID:38165050]. The protein contains three conserved methyltransferase motifs shared across prokaryotes and eukaryotes [PMID:12063391]. Reported roles in chemoresistance and tumor cell behavior have not been connected to the U6 methyltransferase activity in the available corpus [PMID:31656051, PMID:39071668].","teleology":[{"year":2001,"claim":"Before any functional assignment, the gene now known as THUMPD2 (C2orf8) needed basic molecular definition, which established it as a candidate methyltransferase.","evidence":"RACE, RT-PCR, Northern blot, FISH and computational domain analysis of the C2orf8 locus","pmids":["12063391"],"confidence":"Medium","gaps":["No enzymatic activity demonstrated","No substrate identified","No structural model of the catalytic motifs"]},{"year":2021,"claim":"Whether THUMPD2 acts within a defined enzymatic module was unknown; it was placed in the TRMT112 methyltransferase interaction network, predicting a TRMT112-dependent activity.","evidence":"SILAC affinity pulldown and co-expression stability assays in cells","pmids":["34948388"],"confidence":"Medium","gaps":["Substrate RNA/protein not identified in this study","Catalytic activity not directly demonstrated","Single lab"]},{"year":2023,"claim":"The substrate and catalytic identity were open; THUMPD2 was shown to be an active m2G methyltransferase that associates with U6 snRNA and depends on TRMT112 for stability and activity.","evidence":"Interaction network mapping, in vitro methyltransferase assays, and demonstrated direct U6 snRNA association in intact cells","pmids":["37283053"],"confidence":"High","gaps":["Exact modified position not pinpointed in this study","Structural basis of U6 recognition not resolved"]},{"year":2024,"claim":"The precise modification site and physiological consequence were unresolved; THUMPD2 was shown to methylate G72 of U6 via specific recognition elements, with knockout disrupting splicing genome-wide and triggering NMD.","evidence":"THUMPD2 knockout in human cells, in vitro assays with U6 recognition-element mutagenesis, transcriptome-wide splicing analysis, and NMD activation readout","pmids":["38165050"],"confidence":"High","gaps":["Structure of the THUMPD2–TRMT112–U6 complex not determined","Whether specific mis-spliced transcripts drive phenotypes is unclear"]},{"year":2024,"claim":"Beyond splicing, phenotypic roles were explored; THUMPD2 manipulation altered proliferation, metastasis, and chemoresistance in cancer cells, but without a molecular link to its U6 methyltransferase activity.","evidence":"siRNA/shRNA knockdown, overexpression, in vivo tumor growth assays, drug-resistance IC50 comparisons, and GO/KEGG pathway analysis across esophageal and ovarian cancer models","pmids":["31656051","39071668"],"confidence":"Low","gaps":["Single-knockdown approaches with limited mechanistic follow-up","No mechanistic connection between m2G/U6 splicing activity and the cancer phenotypes","Phenotypic readouts only"]},{"year":null,"claim":"It remains unknown how the THUMPD2-dependent U6 m2G72 modification mechanistically links to its reported roles in tumor proliferation, metastasis, and drug resistance.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of the THUMPD2–TRMT112–U6 complex","Cancer phenotypes not tied to splicing function","Disease/Mendelian relevance uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140098","term_label":"catalytic activity, acting on RNA","supporting_discovery_ids":[0,1]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1,3]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[0,1]}],"localization":[],"pathway":[{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[0,1]}],"complexes":[],"partners":["TRMT112"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BTF0","full_name":"U6 snRNA (guanine-N(2))-methyltransferase THUMPD2","aliases":["THUMP domain-containing protein 2"],"length_aa":503,"mass_kda":56.4,"function":"Catalytic subunit of the THUMPD2-TRM112 methyltransferase complex, that specifically mediates the S-adenosyl-L-methionine-dependent N(2)-methylation of guanosine nucleotides, most probably at position 72 (m2G72), in the U6snRNA of the major spliceosome (PubMed:37283053). This modification in the U6 snRNA affects the constitutive splicing efficiency of introns that have suboptimal splice sites and can impact final mRNA levels (PubMed:37283053)","subcellular_location":"Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9BTF0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/THUMPD2","classification":"Not Classified","n_dependent_lines":8,"n_total_lines":1208,"dependency_fraction":0.006622516556291391},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/THUMPD2","total_profiled":1310},"omim":[{"mim_id":"621532","title":"THUMP DOMAIN PROTEIN 3, tRNA GUANOSINE METHYLTRANSFERASE; THUMPD3","url":"https://www.omim.org/entry/621532"},{"mim_id":"618630","title":"tRNA METHYLTRANSFERASE SUBUNIT 11-2; TRMT112","url":"https://www.omim.org/entry/618630"},{"mim_id":"611751","title":"THUMP DOMAIN PROTEIN 2, tRNA and snRNA GUANOSINE METHYLTRANSFERASE; THUMPD2","url":"https://www.omim.org/entry/611751"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Uncertain","locations":[{"location":"Nucleoplasm","reliability":"Uncertain"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/THUMPD2"},"hgnc":{"alias_symbol":["MGC2454"],"prev_symbol":["C2orf8"]},"alphafold":{"accession":"Q9BTF0","domains":[{"cath_id":"3.40.50.150","chopping":"285-420_474-498","consensus_level":"high","plddt":92.5528,"start":285,"end":498}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BTF0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BTF0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BTF0-F1-predicted_aligned_error_v6.png","plddt_mean":75.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=THUMPD2","jax_strain_url":"https://www.jax.org/strain/search?query=THUMPD2"},"sequence":{"accession":"Q9BTF0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BTF0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BTF0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BTF0"}},"corpus_meta":[{"pmid":"35685361","id":"PMC_35685361","title":"Integrative RNA profiling of TBEV-infected neurons and astrocytes reveals potential pathogenic effectors.","date":"2022","source":"Computational and structural biotechnology journal","url":"https://pubmed.ncbi.nlm.nih.gov/35685361","citation_count":31,"is_preprint":false},{"pmid":"37283053","id":"PMC_37283053","title":"N 2-methylguanosine modifications on human tRNAs and snRNA U6 are important for cell proliferation, protein translation and pre-mRNA splicing.","date":"2023","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/37283053","citation_count":29,"is_preprint":false},{"pmid":"34948388","id":"PMC_34948388","title":"Human TRMT112-Methyltransferase Network Consists of Seven Partners Interacting with a Common Co-Factor.","date":"2021","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/34948388","citation_count":26,"is_preprint":false},{"pmid":"38165050","id":"PMC_38165050","title":"THUMPD2 catalyzes the N2-methylation of U6 snRNA of the spliceosome catalytic center and regulates pre-mRNA splicing and retinal degeneration.","date":"2024","source":"Nucleic acids research","url":"https://pubmed.ncbi.nlm.nih.gov/38165050","citation_count":9,"is_preprint":false},{"pmid":"12063391","id":"PMC_12063391","title":"Localization, genomic organization, and alternative transcription of a novel human SAM-dependent methyltransferase gene on chromosome 2p22-->p21.","date":"2001","source":"Cytogenetics and cell genetics","url":"https://pubmed.ncbi.nlm.nih.gov/12063391","citation_count":6,"is_preprint":false},{"pmid":"31656051","id":"PMC_31656051","title":"THUMP domain containing 2 protein possibly induces resistance to cisplatin and 5-fluorouracil in in vitro human esophageal squamous cell carcinoma cells as revealed by transposon activation mutagenesis.","date":"2019","source":"The journal of gene medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31656051","citation_count":5,"is_preprint":false},{"pmid":"39071668","id":"PMC_39071668","title":"RNA-binding protein THUMPD2 inhibits proliferation and promotes metastasis in epithelial ovarian cancer.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/39071668","citation_count":2,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5724,"output_tokens":1606,"usd":0.020631,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8387,"output_tokens":1974,"usd":0.045643,"stage2_stop_reason":"end_turn"},"total_usd":0.066274,"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\": 2023,\n      \"finding\": \"THUMPD2 is an active N2-methylguanosine (m2G) methyltransferase that directly associates with U6 snRNA and is required for formation of the m2G modification on U6 snRNA (the last 'orphan' modification in U6 snRNA). THUMPD2 interacts with the co-factor TRMT112, which is required for its metabolic stability and enzymatic activity.\",\n      \"method\": \"Interaction network mapping via SILAC/pulldown in intact cells; in vitro methyltransferase activity assays; direct association with U6 snRNA demonstrated experimentally\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — enzymatic activity established in vitro, replicated independently in two separate papers (PMID:37283053 and PMID:38165050) using orthogonal methods\",\n      \"pmids\": [\"37283053\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"THUMPD2 is the methyltransferase responsible for N2-methylation of G72 of U6 snRNA (m2G72) by explicitly recognizing U6-specific sequences and structural elements. Knockout of THUMPD2 eliminates U6 m2G72 and impairs pre-mRNA splicing activity, causing thousands of altered alternative splicing events in human cells and activating the nonsense-mediated mRNA decay pathway. The m2G modification at G72 of U6 snRNA is conserved among vertebrates.\",\n      \"method\": \"THUMPD2 knockout in human cells; in vitro methyltransferase assays with mutagenesis of U6 recognition elements; transcriptome-wide alternative splicing analysis; NMD pathway activation assayed\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — enzymatic mechanism established by knockout + in vitro reconstitution with specificity mutagenesis, replicated across two independent studies\",\n      \"pmids\": [\"38165050\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"THUMPD2 was identified as one of seven methyltransferase interaction partners of TRMT112 in a SILAC pulldown screen. TRMT112 stabilizes THUMPD2 in cells, and TRMT112 and THUMPD2 exhibit a strong mutual feedback loop when co-expressed.\",\n      \"method\": \"SILAC-based affinity pulldown; co-expression stability assays; single amino acid mutations on TRMT112 surface to probe interaction differences\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — SILAC pulldown with co-expression validation, single lab, consistent with independent replication in PMID:37283053\",\n      \"pmids\": [\"34948388\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"THUMPD2 (C2orf8) encodes a putative 244 amino acid protein containing three methyltransferase motifs conserved across prokaryotic and eukaryotic species, and is expressed in multiple tissues. The gene was localized to chromosome 2p22→p21 by FISH. Alternative transcripts share a coding region spanning exons 6–10.\",\n      \"method\": \"RACE analysis, RT-PCR, Northern blot, FISH, computational ORF and domain analysis\",\n      \"journal\": \"Cytogenetics and cell genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — genomic localization and domain identification by multiple methods, but functional enzymatic activity not experimentally confirmed in this study\",\n      \"pmids\": [\"12063391\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Downregulation of THUMPD2 expression (via transposon insertion) and siRNA-mediated knockdown of THUMPD2 conferred resistance to both cisplatin and 5-fluorouracil in human esophageal squamous cell carcinoma cells in vitro.\",\n      \"method\": \"Transposon activation mutagenesis screen; siRNA knockdown; IC50 comparison between drug-resistant and wild-type cells\",\n      \"journal\": \"The journal of gene medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single knockdown approach with limited mechanistic follow-up; no pathway placement for the drug resistance phenotype\",\n      \"pmids\": [\"31656051\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"shRNA-mediated knockdown of THUMPD2 in ovarian cancer cell lines (OVCAR3, SKOV3) increased cell proliferation but inhibited metastasis, while THUMPD2 overexpression had the opposite effect, with THUMPD2 overexpression suppressing tumor growth in vivo. GO/KEGG pathway analysis of THUMPD2 target genes implicated centrosome, microtubule, cell cycle, and extracellular matrix pathways.\",\n      \"method\": \"shRNA knockdown; overexpression; in vivo tumor growth assay; GO/KEGG analysis of transcriptome data\",\n      \"journal\": \"Heliyon\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, phenotypic readout without direct molecular mechanism established for THUMPD2's role in proliferation/metastasis\",\n      \"pmids\": [\"39071668\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"THUMPD2 is a TRMT112-dependent N2-methylguanosine (m2G) methyltransferase that specifically recognizes U6 snRNA sequence and structural elements to install the m2G modification at position G72 of U6 snRNA (the catalytic center of the spliceosome), thereby promoting efficient pre-mRNA splicing; loss of THUMPD2 eliminates this modification and globally disrupts alternative splicing, triggering nonsense-mediated mRNA decay.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"THUMPD2 is an RNA methyltransferase that installs the N2-methylguanosine (m2G) modification on U6 small nuclear RNA, thereby promoting spliceosome function and accurate pre-mRNA splicing [#0, #1]. It directly associates with U6 snRNA and catalyzes m2G specifically at position G72, recognizing U6-specific sequence and structural elements, and its activity and metabolic stability depend on the co-factor TRMT112 [#0, #1]. Loss of THUMPD2 eliminates U6 m2G72, alters thousands of alternative splicing events, and activates the nonsense-mediated mRNA decay pathway, establishing this single modification as functionally important for splicing fidelity [#1]. The protein contains three conserved methyltransferase motifs shared across prokaryotes and eukaryotes [#3]. Reported roles in chemoresistance and tumor cell behavior have not been connected to the U6 methyltransferase activity in the available corpus [#4, #5].\"\n  ,\n  \"teleology\": [\n    {\n      \"year\": 2001,\n      \"claim\": \"Before any functional assignment, the gene now known as THUMPD2 (C2orf8) needed basic molecular definition, which established it as a candidate methyltransferase.\",\n      \"evidence\": \"RACE, RT-PCR, Northern blot, FISH and computational domain analysis of the C2orf8 locus\",\n      \"pmids\": [\"12063391\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No enzymatic activity demonstrated\", \"No substrate identified\", \"No structural model of the catalytic motifs\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Whether THUMPD2 acts within a defined enzymatic module was unknown; it was placed in the TRMT112 methyltransferase interaction network, predicting a TRMT112-dependent activity.\",\n      \"evidence\": \"SILAC affinity pulldown and co-expression stability assays in cells\",\n      \"pmids\": [\"34948388\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Substrate RNA/protein not identified in this study\", \"Catalytic activity not directly demonstrated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"The substrate and catalytic identity were open; THUMPD2 was shown to be an active m2G methyltransferase that associates with U6 snRNA and depends on TRMT112 for stability and activity.\",\n      \"evidence\": \"Interaction network mapping, in vitro methyltransferase assays, and demonstrated direct U6 snRNA association in intact cells\",\n      \"pmids\": [\"37283053\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Exact modified position not pinpointed in this study\", \"Structural basis of U6 recognition not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"The precise modification site and physiological consequence were unresolved; THUMPD2 was shown to methylate G72 of U6 via specific recognition elements, with knockout disrupting splicing genome-wide and triggering NMD.\",\n      \"evidence\": \"THUMPD2 knockout in human cells, in vitro assays with U6 recognition-element mutagenesis, transcriptome-wide splicing analysis, and NMD activation readout\",\n      \"pmids\": [\"38165050\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structure of the THUMPD2–TRMT112–U6 complex not determined\", \"Whether specific mis-spliced transcripts drive phenotypes is unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Beyond splicing, phenotypic roles were explored; THUMPD2 manipulation altered proliferation, metastasis, and chemoresistance in cancer cells, but without a molecular link to its U6 methyltransferase activity.\",\n      \"evidence\": \"siRNA/shRNA knockdown, overexpression, in vivo tumor growth assays, drug-resistance IC50 comparisons, and GO/KEGG pathway analysis across esophageal and ovarian cancer models\",\n      \"pmids\": [\"31656051\", \"39071668\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single-knockdown approaches with limited mechanistic follow-up\", \"No mechanistic connection between m2G/U6 splicing activity and the cancer phenotypes\", \"Phenotypic readouts only\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how the THUMPD2-dependent U6 m2G72 modification mechanistically links to its reported roles in tumor proliferation, metastasis, and drug resistance.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of the THUMPD2–TRMT112–U6 complex\", \"Cancer phenotypes not tied to splicing function\", \"Disease/Mendelian relevance uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140098\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0008168\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 3]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [0, 1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"TRMT112\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}