{"gene":"GID8","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":2003,"finding":"Twa1 (GID8) was identified as a novel nuclear protein that forms a protein complex with RanBPM and hMuskelin, demonstrated by two-hybrid screening, immunoprecipitation, and gel-filtration analyses. Twa1 contains LisH-CTLH motifs and localizes within the nucleus.","method":"Yeast two-hybrid screening, co-immunoprecipitation, gel-filtration chromatography","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP and gel filtration in single lab with two orthogonal methods","pmids":["12559565"],"is_preprint":false},{"year":2017,"finding":"TWA1/GID8 constitutively forms stable homodimers via its LisH-CTLH-CRA domains, as shown by gel filtration, SEC-MALS, and native PAGE with recombinant tag-free mouse TWA1; dimerization is independent of its single cysteine residue (Cys139) and is not disulfide-mediated. CD spectroscopy showed it is predominantly α-helical.","method":"Gel filtration chromatography, SEC-MALS, native PAGE, CD spectroscopy, recombinant protein purification","journal":"Bioscience reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple orthogonal biophysical methods (SEC-MALS, native PAGE, CD) on recombinant protein with mutagenesis-informed cysteine control in one rigorous study","pmids":["27920276"],"is_preprint":false},{"year":2017,"finding":"NMR chemical shift assignments of the LisH domain homodimer of human Twa1/GID8 were determined, confirming homodimerization of the LisH domain in solution and providing the first step toward its solution structure.","method":"NMR spectroscopy (1H, 13C, 15N chemical shift assignment)","journal":"Biomolecular NMR assignments","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — NMR structural assignment in single lab, no full structure or functional mutagenesis reported","pmids":["29067546"],"is_preprint":false},{"year":2017,"finding":"Twa1/GID8 acts as a nuclear retention factor for β-catenin in Wnt signaling. In the absence of Wnt, Twa1 associates with β-catenin in the Axin destruction complex and undergoes ubiquitination and degradation. Upon Wnt activation, Twa1 translocates into the nucleus where it binds and retains β-catenin. Depletion of Twa1 attenuates Wnt-stimulated gene expression, dorsal development in zebrafish embryos, and xenograft tumor growth of CRC cells.","method":"Co-immunoprecipitation, siRNA/shRNA knockdown, reporter gene assays, zebrafish embryo epistasis, xenograft tumor assay, nuclear/cytoplasmic fractionation","journal":"Cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (Co-IP, genetic epistasis in zebrafish, xenograft, reporter assays) demonstrating nuclear retention mechanism, single lab but with in vivo validation","pmids":["28829046"],"is_preprint":false},{"year":2024,"finding":"GID8/Twa1 maintains active glutamine uptake and metabolism by regulating expression of excitatory amino acid transporter SLC1A3 and glutaminase (GLS), facilitating CRC progression. YTHDF1 promotes GID8 translation efficiency in an m6A-dependent manner.","method":"In vitro knockdown/overexpression assays, in vivo xenograft, m6A-dependent translation reporter, metabolic assays","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — functional knockdown with defined metabolic readout (SLC1A3, GLS regulation) and in vivo validation, single lab","pmids":["39151722"],"is_preprint":false},{"year":2026,"finding":"Drosophila Pasovec (Psv; mammalian Gid8 ortholog) is required for nuclear translocation of β-catenin/Armadillo in Wg/Wnt signaling. Psv physically binds IFT140, a core component of the IFT-A complex, independently of Wg/Wnt activation. The CRA domain of Psv/Gid8 mediates its own nuclear localization and that of β-catenin/Arm; CRA domain mutations abolish nuclear localization of both Psv and β-catenin, and a CRA-mutant Psv acts as a dominant inhibitor of Wg/Wnt signaling.","method":"Drosophila loss-of-function genetics, co-immunoprecipitation (Psv-IFT140 interaction), domain mutagenesis (CRA domain mutants), epistasis analysis, fluorescence imaging of β-catenin/Arm localization","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, domain mutagenesis with defined phenotypic readout, genetic epistasis in Drosophila, multiple orthogonal methods in one study","pmids":["41931626"],"is_preprint":false}],"current_model":"GID8/Twa1 is a conserved LisH-CTLH-CRA domain protein that constitutively homodimerizes and functions as a nuclear retention factor for β-catenin in Wnt/Wg signaling: upon pathway activation it translocates to the nucleus (via its CRA domain), binds and retains β-catenin there, and physically associates with the IFT-A complex component IFT140 to facilitate β-catenin nuclear import; in the absence of Wnt it is part of the Axin destruction complex and undergoes ubiquitination/degradation, and it also promotes colorectal cancer progression by regulating glutamine metabolism through SLC1A3 and GLS."},"narrative":{"mechanistic_narrative":"GID8/Twa1 is a conserved LisH-CTLH-CRA domain protein that functions as a nuclear retention factor for β-catenin in Wnt/Wg signaling [PMID:28829046]. It constitutively forms stable, predominantly α-helical homodimers through its LisH-CTLH-CRA domains, independently of disulfide bonding [PMID:27920276, PMID:29067546], and assembles into a nuclear complex with RanBPM and hMuskelin [PMID:12559565]. In the absence of Wnt, Twa1 associates with β-catenin in the Axin destruction complex and is ubiquitinated and degraded; upon Wnt activation it translocates into the nucleus where it binds and retains β-catenin, and its depletion attenuates Wnt-stimulated gene expression, dorsal development in zebrafish, and CRC xenograft growth [PMID:28829046]. Nuclear translocation of both Twa1 and β-catenin is governed by the CRA domain and requires physical association with the IFT-A component IFT140, which Twa1 binds independently of pathway activation; CRA-domain mutants fail to enter the nucleus and act as dominant inhibitors of Wg/Wnt signaling [PMID:41931626]. Beyond Wnt signaling, GID8 sustains glutamine uptake and metabolism by regulating SLC1A3 and glutaminase (GLS) expression to drive colorectal cancer progression, and its own translation is enhanced by YTHDF1 in an m6A-dependent manner [PMID:39151722].","teleology":[{"year":2003,"claim":"Established GID8/Twa1 as a nuclear protein embedded in a defined protein complex, giving the first clue to its molecular context.","evidence":"Yeast two-hybrid, reciprocal Co-IP, and gel filtration identifying a RanBPM/hMuskelin complex","pmids":["12559565"],"confidence":"Medium","gaps":["Functional role of the complex undefined","No link to a signaling pathway yet","LisH-CTLH motif function not tested"]},{"year":2017,"claim":"Resolved the oligomeric state of the protein, showing it is an obligate homodimer assembled through its LisH-CTLH-CRA domains rather than transient or disulfide-linked association.","evidence":"SEC-MALS, native PAGE, CD spectroscopy on recombinant tag-free mouse TWA1 with Cys139 control; NMR chemical-shift assignment of the human LisH domain homodimer","pmids":["27920276","29067546"],"confidence":"High","gaps":["No full solution or crystal structure","Functional consequence of dimerization for partner binding untested"]},{"year":2017,"claim":"Defined the central mechanistic role: Twa1 is a Wnt-regulated nuclear retention factor for β-catenin, linking its degradation in the Axin complex to nuclear accumulation upon pathway activation.","evidence":"Co-IP, siRNA/shRNA knockdown, reporter assays, nuclear/cytoplasmic fractionation, zebrafish epistasis, and CRC xenografts","pmids":["28829046"],"confidence":"High","gaps":["Mechanism of Wnt-triggered nuclear translocation not resolved at this stage","Identity of the responsible ubiquitin ligase unspecified","Domain requirements not mapped"]},{"year":2024,"claim":"Extended GID8 function beyond Wnt to metabolic control, showing it drives glutamine uptake and CRC progression via SLC1A3/GLS, and that its expression is set by m6A-dependent translation.","evidence":"Knockdown/overexpression, metabolic assays, in vivo xenograft, and m6A-dependent translation reporter (YTHDF1)","pmids":["39151722"],"confidence":"Medium","gaps":["Direct vs indirect regulation of SLC1A3/GLS unclear","Relationship between metabolic and Wnt roles undefined","Single lab without orthogonal confirmation"]},{"year":2026,"claim":"Mapped the translocation mechanism to the CRA domain and identified IFT140 as the physical partner enabling β-catenin nuclear import, explaining how Twa1 ferries β-catenin into the nucleus.","evidence":"Drosophila loss-of-function genetics, reciprocal Co-IP (Psv-IFT140), CRA domain mutagenesis, epistasis, and β-catenin/Arm localization imaging","pmids":["41931626"],"confidence":"High","gaps":["Structural basis of CRA-mediated nuclear entry unknown","How IFT-A component mediates import mechanistically unresolved","Conservation of IFT140 dependence in mammalian CRC not directly tested"]},{"year":null,"claim":"How the homodimeric LisH-CTLH-CRA architecture, Axin-complex degradation, IFT140-dependent import, and glutamine metabolic control are mechanistically integrated remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No structure of the Twa1-β-catenin or Twa1-IFT140 complex","Ubiquitin ligase acting on Twa1 unidentified","Causal connection between Wnt and metabolic functions unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[3,5]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,3,5]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3,5]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[4]}],"complexes":["Axin destruction complex"],"partners":["CTNNB1","IFT140","RANBPM","MKLN1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NWU2","full_name":"Glucose-induced degradation protein 8 homolog","aliases":["Two hybrid-associated protein 1 with RanBPM","Twa1"],"length_aa":228,"mass_kda":26.7,"function":"Core component of the CTLH E3 ubiquitin-protein ligase complex that selectively accepts ubiquitin from UBE2H and mediates ubiquitination and subsequent proteasomal degradation of the transcription factor HBP1 (PubMed:29911972). Acts as a positive regulator of Wnt signaling pathway by promoting beta-catenin (CTNNB1) nuclear accumulation (PubMed:28829046)","subcellular_location":"Cytoplasm; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9NWU2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GID8","classification":"Not Classified","n_dependent_lines":25,"n_total_lines":1208,"dependency_fraction":0.020695364238410598},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"RANBP10","stoichiometry":10.0},{"gene":"RANBP9","stoichiometry":10.0},{"gene":"HNRNPH1","stoichiometry":0.2},{"gene":"SNRPF","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/GID8","total_profiled":1310},"omim":[{"mim_id":"611625","title":"GID COMPLEX, SUBUNIT 8; GID8","url":"https://www.omim.org/entry/611625"},{"mim_id":"605623","title":"MUSKELIN 1; MKLN1","url":"https://www.omim.org/entry/605623"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Cell Junctions","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/GID8"},"hgnc":{"alias_symbol":["FLJ20602","bA305P22.1","TWA1"],"prev_symbol":["C20orf11"]},"alphafold":{"accession":"Q9NWU2","domains":[{"cath_id":"-","chopping":"64-186","consensus_level":"high","plddt":93.4144,"start":64,"end":186}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NWU2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NWU2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NWU2-F1-predicted_aligned_error_v6.png","plddt_mean":91.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GID8","jax_strain_url":"https://www.jax.org/strain/search?query=GID8"},"sequence":{"accession":"Q9NWU2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NWU2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NWU2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NWU2"}},"corpus_meta":[{"pmid":"12559565","id":"PMC_12559565","title":"A novel nuclear protein, Twa1, and Muskelin comprise a complex with RanBPM.","date":"2003","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/12559565","citation_count":62,"is_preprint":false},{"pmid":"28829046","id":"PMC_28829046","title":"Twa1/Gid8 is a β-catenin nuclear retention factor in Wnt signaling and colorectal tumorigenesis.","date":"2017","source":"Cell research","url":"https://pubmed.ncbi.nlm.nih.gov/28829046","citation_count":52,"is_preprint":false},{"pmid":"34382916","id":"PMC_34382916","title":"circ_C20orf11 enhances DDP resistance by inhibiting miR-527/YWHAZ through the promotion of extracellular vesicle-mediated macrophage M2 polarization in ovarian cancer.","date":"2021","source":"Cancer biology & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/34382916","citation_count":27,"is_preprint":false},{"pmid":"39151722","id":"PMC_39151722","title":"YTHDF1 regulates GID8-mediated glutamine metabolism to promote colorectal cancer progression in m6A-dependent manner.","date":"2024","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/39151722","citation_count":20,"is_preprint":false},{"pmid":"27920276","id":"PMC_27920276","title":"Studies of recombinant TWA1 reveal constitutive dimerization.","date":"2017","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/27920276","citation_count":7,"is_preprint":false},{"pmid":"29067546","id":"PMC_29067546","title":"1H, 13C and 15N chemical shift assignment of lissencephaly-1 homology (LisH) domain homodimer of human two-hybrid-associated protein 1 with RanBPM (Twa1).","date":"2017","source":"Biomolecular NMR assignments","url":"https://pubmed.ncbi.nlm.nih.gov/29067546","citation_count":2,"is_preprint":false},{"pmid":"40350652","id":"PMC_40350652","title":"circ-C20orf11 promotes karyopherin alpha 2 expression through miR-495-3p and affects non-small cell lung cancer development.","date":"2025","source":"Journal of physiology and pharmacology : an official journal of the Polish Physiological Society","url":"https://pubmed.ncbi.nlm.nih.gov/40350652","citation_count":0,"is_preprint":false},{"pmid":"41931626","id":"PMC_41931626","title":"Nuclear translocation of β-catenin in Wg/Wnt signaling via the IFT-A microtubule-associated complex requires Pasovec/Gid8 proteins.","date":"2026","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/41931626","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":6052,"output_tokens":1713,"usd":0.021926,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8513,"output_tokens":2219,"usd":0.04902,"stage2_stop_reason":"end_turn"},"total_usd":0.070946,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"Twa1 (GID8) was identified as a novel nuclear protein that forms a protein complex with RanBPM and hMuskelin, demonstrated by two-hybrid screening, immunoprecipitation, and gel-filtration analyses. Twa1 contains LisH-CTLH motifs and localizes within the nucleus.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation, gel-filtration chromatography\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP and gel filtration in single lab with two orthogonal methods\",\n      \"pmids\": [\"12559565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"TWA1/GID8 constitutively forms stable homodimers via its LisH-CTLH-CRA domains, as shown by gel filtration, SEC-MALS, and native PAGE with recombinant tag-free mouse TWA1; dimerization is independent of its single cysteine residue (Cys139) and is not disulfide-mediated. CD spectroscopy showed it is predominantly α-helical.\",\n      \"method\": \"Gel filtration chromatography, SEC-MALS, native PAGE, CD spectroscopy, recombinant protein purification\",\n      \"journal\": \"Bioscience reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple orthogonal biophysical methods (SEC-MALS, native PAGE, CD) on recombinant protein with mutagenesis-informed cysteine control in one rigorous study\",\n      \"pmids\": [\"27920276\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NMR chemical shift assignments of the LisH domain homodimer of human Twa1/GID8 were determined, confirming homodimerization of the LisH domain in solution and providing the first step toward its solution structure.\",\n      \"method\": \"NMR spectroscopy (1H, 13C, 15N chemical shift assignment)\",\n      \"journal\": \"Biomolecular NMR assignments\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — NMR structural assignment in single lab, no full structure or functional mutagenesis reported\",\n      \"pmids\": [\"29067546\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Twa1/GID8 acts as a nuclear retention factor for β-catenin in Wnt signaling. In the absence of Wnt, Twa1 associates with β-catenin in the Axin destruction complex and undergoes ubiquitination and degradation. Upon Wnt activation, Twa1 translocates into the nucleus where it binds and retains β-catenin. Depletion of Twa1 attenuates Wnt-stimulated gene expression, dorsal development in zebrafish embryos, and xenograft tumor growth of CRC cells.\",\n      \"method\": \"Co-immunoprecipitation, siRNA/shRNA knockdown, reporter gene assays, zebrafish embryo epistasis, xenograft tumor assay, nuclear/cytoplasmic fractionation\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (Co-IP, genetic epistasis in zebrafish, xenograft, reporter assays) demonstrating nuclear retention mechanism, single lab but with in vivo validation\",\n      \"pmids\": [\"28829046\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"GID8/Twa1 maintains active glutamine uptake and metabolism by regulating expression of excitatory amino acid transporter SLC1A3 and glutaminase (GLS), facilitating CRC progression. YTHDF1 promotes GID8 translation efficiency in an m6A-dependent manner.\",\n      \"method\": \"In vitro knockdown/overexpression assays, in vivo xenograft, m6A-dependent translation reporter, metabolic assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — functional knockdown with defined metabolic readout (SLC1A3, GLS regulation) and in vivo validation, single lab\",\n      \"pmids\": [\"39151722\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Drosophila Pasovec (Psv; mammalian Gid8 ortholog) is required for nuclear translocation of β-catenin/Armadillo in Wg/Wnt signaling. Psv physically binds IFT140, a core component of the IFT-A complex, independently of Wg/Wnt activation. The CRA domain of Psv/Gid8 mediates its own nuclear localization and that of β-catenin/Arm; CRA domain mutations abolish nuclear localization of both Psv and β-catenin, and a CRA-mutant Psv acts as a dominant inhibitor of Wg/Wnt signaling.\",\n      \"method\": \"Drosophila loss-of-function genetics, co-immunoprecipitation (Psv-IFT140 interaction), domain mutagenesis (CRA domain mutants), epistasis analysis, fluorescence imaging of β-catenin/Arm localization\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, domain mutagenesis with defined phenotypic readout, genetic epistasis in Drosophila, multiple orthogonal methods in one study\",\n      \"pmids\": [\"41931626\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"GID8/Twa1 is a conserved LisH-CTLH-CRA domain protein that constitutively homodimerizes and functions as a nuclear retention factor for β-catenin in Wnt/Wg signaling: upon pathway activation it translocates to the nucleus (via its CRA domain), binds and retains β-catenin there, and physically associates with the IFT-A complex component IFT140 to facilitate β-catenin nuclear import; in the absence of Wnt it is part of the Axin destruction complex and undergoes ubiquitination/degradation, and it also promotes colorectal cancer progression by regulating glutamine metabolism through SLC1A3 and GLS.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"GID8/Twa1 is a conserved LisH-CTLH-CRA domain protein that functions as a nuclear retention factor for β-catenin in Wnt/Wg signaling [#3]. It constitutively forms stable, predominantly α-helical homodimers through its LisH-CTLH-CRA domains, independently of disulfide bonding [#1, #2], and assembles into a nuclear complex with RanBPM and hMuskelin [#0]. In the absence of Wnt, Twa1 associates with β-catenin in the Axin destruction complex and is ubiquitinated and degraded; upon Wnt activation it translocates into the nucleus where it binds and retains β-catenin, and its depletion attenuates Wnt-stimulated gene expression, dorsal development in zebrafish, and CRC xenograft growth [#3]. Nuclear translocation of both Twa1 and β-catenin is governed by the CRA domain and requires physical association with the IFT-A component IFT140, which Twa1 binds independently of pathway activation; CRA-domain mutants fail to enter the nucleus and act as dominant inhibitors of Wg/Wnt signaling [#5]. Beyond Wnt signaling, GID8 sustains glutamine uptake and metabolism by regulating SLC1A3 and glutaminase (GLS) expression to drive colorectal cancer progression, and its own translation is enhanced by YTHDF1 in an m6A-dependent manner [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established GID8/Twa1 as a nuclear protein embedded in a defined protein complex, giving the first clue to its molecular context.\",\n      \"evidence\": \"Yeast two-hybrid, reciprocal Co-IP, and gel filtration identifying a RanBPM/hMuskelin complex\",\n      \"pmids\": [\"12559565\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional role of the complex undefined\", \"No link to a signaling pathway yet\", \"LisH-CTLH motif function not tested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Resolved the oligomeric state of the protein, showing it is an obligate homodimer assembled through its LisH-CTLH-CRA domains rather than transient or disulfide-linked association.\",\n      \"evidence\": \"SEC-MALS, native PAGE, CD spectroscopy on recombinant tag-free mouse TWA1 with Cys139 control; NMR chemical-shift assignment of the human LisH domain homodimer\",\n      \"pmids\": [\"27920276\", \"29067546\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No full solution or crystal structure\", \"Functional consequence of dimerization for partner binding untested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined the central mechanistic role: Twa1 is a Wnt-regulated nuclear retention factor for β-catenin, linking its degradation in the Axin complex to nuclear accumulation upon pathway activation.\",\n      \"evidence\": \"Co-IP, siRNA/shRNA knockdown, reporter assays, nuclear/cytoplasmic fractionation, zebrafish epistasis, and CRC xenografts\",\n      \"pmids\": [\"28829046\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism of Wnt-triggered nuclear translocation not resolved at this stage\", \"Identity of the responsible ubiquitin ligase unspecified\", \"Domain requirements not mapped\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended GID8 function beyond Wnt to metabolic control, showing it drives glutamine uptake and CRC progression via SLC1A3/GLS, and that its expression is set by m6A-dependent translation.\",\n      \"evidence\": \"Knockdown/overexpression, metabolic assays, in vivo xenograft, and m6A-dependent translation reporter (YTHDF1)\",\n      \"pmids\": [\"39151722\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect regulation of SLC1A3/GLS unclear\", \"Relationship between metabolic and Wnt roles undefined\", \"Single lab without orthogonal confirmation\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Mapped the translocation mechanism to the CRA domain and identified IFT140 as the physical partner enabling β-catenin nuclear import, explaining how Twa1 ferries β-catenin into the nucleus.\",\n      \"evidence\": \"Drosophila loss-of-function genetics, reciprocal Co-IP (Psv-IFT140), CRA domain mutagenesis, epistasis, and β-catenin/Arm localization imaging\",\n      \"pmids\": [\"41931626\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of CRA-mediated nuclear entry unknown\", \"How IFT-A component mediates import mechanistically unresolved\", \"Conservation of IFT140 dependence in mammalian CRC not directly tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the homodimeric LisH-CTLH-CRA architecture, Axin-complex degradation, IFT140-dependent import, and glutamine metabolic control are mechanistically integrated remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structure of the Twa1-β-catenin or Twa1-IFT140 complex\", \"Ubiquitin ligase acting on Twa1 unidentified\", \"Causal connection between Wnt and metabolic functions unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 3, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3, 5]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"complexes\": [\"Axin destruction complex\"],\n    \"partners\": [\"CTNNB1\", \"IFT140\", \"RanBPM\", \"MKLN1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":5,"faith_pct":80.0}}