{"gene":"MIOS","run_date":"2026-06-10T02:59:50","timeline":{"discoveries":[{"year":2024,"finding":"Rbpms2 acts upstream of the GATOR2 complex component Mios (MIOS) to promote oocyte fate in zebrafish; genetic analyses indicate Rbpms2 promotes nucleolar amplification via the mTORC1 signaling pathway specifically through Mios, placing Mios as an essential GATOR2-mediated checkpoint component in oogenesis.","method":"Genetic epistasis analysis in zebrafish; translational regulation assays; loss-of-function","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with defined pathway placement in zebrafish, single lab, multiple genetic tools but no in vitro reconstitution","pmids":["38898112"],"is_preprint":false},{"year":2024,"finding":"Rbpms2 acts upstream of Mios (MIOS/GATOR2) to promote mTORC1 activation and nucleolar amplification during oogenesis in zebrafish, consistent with Mios functioning as a GATOR2 component that activates mTORC1 signaling in a nutrient-sensing pathway context.","method":"Genetic epistasis in zebrafish; loss-of-function","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 2 / Weak — preprint, single lab, overlapping with published version; included as corroboration","pmids":["38328218"],"is_preprint":true},{"year":2024,"finding":"MIOS (GATOR2 component) is required for Tanshinone IIA (T2A)-mediated inhibition of mTORC1 and induction of autophagy; in Dictyostelium discoideum, ablation of mios abolished T2A-induced autophagy and mTORC1 inhibition, placing MIOS downstream of sestrin 2 (SESN) in the SESN–GATOR2–mTORC1 axis.","method":"Genetic ablation (mios- cells) in Dictyostelium discoideum; pharmacological treatment; mTORC1 activity assay; autophagy assay","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean genetic loss-of-function with specific cellular readouts (mTORC1 activity, autophagy), single lab, two orthogonal readouts","pmids":["38928292"],"is_preprint":false},{"year":2021,"finding":"YULINK (MIOS) knockdown in mouse HL-1 cardiomyocytes reduced DNA binding activity of PPARγ and decreased SERCA2 expression; PPARγ agonists rescued SERCA2 expression in YULINK-KD cells and PPARγ shRNA knockdown phenocopied YULINK loss, indicating YULINK regulates SERCA2 expression through PPARγ nuclear entry.","method":"shRNA knockdown in mouse HL-1 cardiomyocytes; DNA binding assay for PPARγ; western blot; agonist rescue; Ca2+ imaging; morpholino knockdown in zebrafish","journal":"Journal of biomedical science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (KD, agonist rescue, PPARγ-shRNA phenocopy, DNA binding assay), single lab","pmids":["33423678"],"is_preprint":false},{"year":2023,"finding":"YULINK (MIOS) co-localizes and physically interacts with endosome-related proteins EPS15, RAB33B, TICAM2 and endocytic markers (Clathrin, RHOB) in HUVECs; YULINK knockdown compromised VEGF-induced VEGFR2 internalization, indicating YULINK participates in endocytic vesicle trafficking to regulate vascular formation.","method":"Yeast two-hybrid; FLIM-FRET; immunoprecipitation; immunofluorescence imaging; gene knockdown in zebrafish and HUVECs","journal":"Biological research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal binding confirmation by yeast two-hybrid and FLIM-FRET plus IP, single lab, multiple orthogonal methods","pmids":["36843032"],"is_preprint":false},{"year":2023,"finding":"YULINK (MIOS) knockdown in PASMCs suppressed PDGFR expression and phosphorylation of FAK, PI3K, and AKT, indicating YULINK acts upstream of the PI3K-AKT signaling pathway to regulate PASMC migration, proliferation, and glycolysis.","method":"shRNA knockdown and overexpression in PASMCs; western blot for pathway components; glucose uptake assay; migration assay; immunofluorescence co-localization","journal":"Biological research","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway placement inferred from inhibitor/KD readouts without direct binding evidence for PI3K-AKT components","pmids":["38057829"],"is_preprint":false},{"year":2025,"finding":"YULINK (MIOS) co-localizes and physically interacts with glucose transporter GLUT1; YULINK knockdown suppressed GLUT1 expression and disrupted GLUT1 translocation from cytosol to cell membrane, resulting in reduced glucose uptake and glycolysis in HCC (Huh7) cells.","method":"Proximity ligation assay; immunoprecipitation; immunofluorescence; glucose uptake assay; glycolytic function assay; western blot; shRNA knockdown","journal":"Molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct physical interaction confirmed by two orthogonal methods (PLA + IP), functional consequence demonstrated, single lab","pmids":["41013205"],"is_preprint":false},{"year":2018,"finding":"YULINK (MIOS) regulates SERCA2 expression through PPARγ in cardiomyocytes; YULINK knockdown reduced PPARγ activity and SERCA2 expression, leading to disrupted Ca2+ reuptake and cardiac arrhythmia phenotypes in zebrafish and mouse cardiomyocytes. NOTE: This paper was retracted.","method":"Morpholino knockdown in zebrafish; shRNA knockdown in mouse cardiomyocytes; PPARγ activity assay; Ca2+ imaging; western blot [RETRACTED paper]","journal":"FASEB journal","confidence":"Low","confidence_rationale":"Tier 2 / Weak — paper was retracted; findings partially corroborated by later non-retracted work (PMID:33423678) but original report cannot be trusted independently","pmids":["29401584"],"is_preprint":false}],"current_model":"MIOS (YULINK/Sea4) is a WD40-repeat-containing subunit of the GATOR2 complex that promotes mTORC1 activation in nutrient-sensing pathways; it physically interacts with GLUT1 to regulate glucose transporter membrane localization and glycolysis, associates with endocytic machinery (EPS15, RAB33B, Clathrin) to support VEGFR2 internalization and vascular formation, and in cardiomyocytes regulates SERCA2 expression via a PPARγ-dependent mechanism to maintain Ca2+ cycling."},"narrative":{"mechanistic_narrative":"MIOS (YULINK/Sea4) is a WD40-repeat subunit of the GATOR2 complex that promotes mTORC1 activation within nutrient-sensing pathways [PMID:38898112, PMID:38928292]. In the SESN–GATOR2–mTORC1 axis, MIOS acts downstream of sestrin 2 and is required for inhibition of mTORC1 and induction of autophagy, since its ablation in Dictyostelium abolished both responses [PMID:38928292]; in zebrafish oogenesis it functions as an essential GATOR2-mediated checkpoint downstream of Rbpms2 to drive mTORC1-dependent nucleolar amplification and oocyte fate [PMID:38898112]. Beyond this canonical role, MIOS engages glucose-handling and trafficking machinery: it physically interacts with the glucose transporter GLUT1 and is required for GLUT1 expression and translocation from cytosol to the plasma membrane, thereby supporting glucose uptake and glycolysis [PMID:41013205], and it associates with endocytic proteins EPS15, RAB33B, TICAM2, Clathrin and RHOB to support VEGF-induced VEGFR2 internalization and vascular formation [PMID:36843032]. In cardiomyocytes, MIOS regulates SERCA2 expression through a PPARγ-dependent mechanism, controlling Ca2+ reuptake [PMID:33423678].","teleology":[{"year":2018,"claim":"An initial question was whether MIOS/YULINK has a role in cardiac calcium handling; loss-of-function linked it to SERCA2 expression and Ca2+ cycling, but the founding report was later retracted.","evidence":"Morpholino knockdown in zebrafish and shRNA in mouse cardiomyocytes with PPARγ activity and Ca2+ imaging (retracted)","pmids":["29401584"],"confidence":"Low","gaps":["Original report retracted; cannot be trusted independently","Mechanism connecting MIOS to PPARγ not defined at the molecular level"]},{"year":2021,"claim":"Re-establishing the cardiac role with non-retracted data clarified that MIOS controls SERCA2 expression specifically through promoting PPARγ nuclear/DNA-binding activity.","evidence":"shRNA knockdown in HL-1 cardiomyocytes with PPARγ DNA-binding assay, agonist rescue, PPARγ-shRNA phenocopy, and Ca2+ imaging","pmids":["33423678"],"confidence":"Medium","gaps":["How MIOS regulates PPARγ activity mechanistically is unresolved","No demonstrated direct physical interaction between MIOS and PPARγ"]},{"year":2023,"claim":"Whether MIOS participates in membrane trafficking was tested in endothelial cells, showing it physically associates with endocytic machinery to enable receptor internalization.","evidence":"Yeast two-hybrid, FLIM-FRET, IP and immunofluorescence in HUVECs, with VEGFR2 internalization assays and zebrafish knockdown","pmids":["36843032"],"confidence":"Medium","gaps":["Direct vs. scaffolding role within the endocytic complex not resolved","Relationship between this trafficking function and GATOR2/mTORC1 role unknown"]},{"year":2023,"claim":"A parallel study placed MIOS upstream of PI3K-AKT signaling in vascular smooth muscle, linking it to migration, proliferation and glycolysis.","evidence":"shRNA knockdown and overexpression in PASMCs with western blot of pathway components, glucose uptake and migration assays","pmids":["38057829"],"confidence":"Low","gaps":["Pathway placement inferred from KD readouts without direct binding evidence for PI3K-AKT components","Mechanism connecting MIOS to PDGFR expression not established"]},{"year":2024,"claim":"The canonical GATOR2/mTORC1 function was established in two systems: MIOS is required downstream of sestrin 2 for mTORC1 inhibition and autophagy, and acts as an mTORC1-activating GATOR2 checkpoint in oocyte fate determination.","evidence":"Genetic ablation in Dictyostelium with mTORC1 and autophagy readouts; genetic epistasis with Rbpms2 in zebrafish oogenesis (plus corroborating preprint)","pmids":["38928292","38898112","38328218"],"confidence":"Medium","gaps":["Structural basis of MIOS within GATOR2 in these systems not addressed","Whether nutrient-sensing and trafficking/cardiac roles are mechanistically connected is unknown"]},{"year":2025,"claim":"Whether MIOS directly controls a glucose transporter was answered by showing physical interaction with GLUT1 and a requirement for GLUT1 membrane translocation, tying MIOS to glycolytic metabolism.","evidence":"Proximity ligation assay and IP for interaction; immunofluorescence, glucose uptake and glycolytic assays in Huh7 HCC cells","pmids":["41013205"],"confidence":"Medium","gaps":["Mechanism by which MIOS drives GLUT1 trafficking not defined","Relationship to MIOS endocytic/GATOR2 functions unresolved"]},{"year":null,"claim":"It remains unknown how MIOS's GATOR2/mTORC1 nutrient-sensing function relates mechanistically to its reported roles in GLUT1 trafficking, endocytic VEGFR2 internalization, and PPARγ-dependent SERCA2 regulation.","evidence":"No timeline study integrates these activities into a single mechanism","pmids":[],"confidence":"Low","gaps":["No structural model of MIOS in human cells","Whether trafficking and metabolic roles are mTORC1-dependent is untested","Direct binding partners bridging these functions not identified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,2]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[4]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[2]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[4]}],"complexes":["GATOR2"],"partners":["GLUT1","EPS15","RAB33B","TICAM2","RHOB"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9NXC5","full_name":"GATOR2 complex protein MIOS","aliases":["Missing oocyte meiosis regulator homolog"],"length_aa":875,"mass_kda":98.6,"function":"As a component of the GATOR2 complex, functions as an activator of the amino acid-sensing branch of the mTORC1 signaling pathway (PubMed:23723238, PubMed:26586190, PubMed:27487210, PubMed:35831510, PubMed:36528027). The GATOR2 complex indirectly activates mTORC1 through the inhibition of the GATOR1 subcomplex (PubMed:23723238, PubMed:26586190, PubMed:27487210, PubMed:35831510, PubMed:36528027). GATOR2 probably acts as an E3 ubiquitin-protein ligase toward GATOR1 (PubMed:36528027). In the presence of abundant amino acids, the GATOR2 complex mediates ubiquitination of the NPRL2 core component of the GATOR1 complex, leading to GATOR1 inactivation (PubMed:36528027). In the absence of amino acids, GATOR2 is inhibited, activating the GATOR1 complex (PubMed:25263562, PubMed:25457612, PubMed:26586190, PubMed:27487210). Within the GATOR2 complex, MIOS is required to prevent autoubiquitination of WDR24, the catalytic subunit of the complex (PubMed:35831510). The GATOR2 complex is required for brain myelination (By similarity)","subcellular_location":"Lysosome membrane","url":"https://www.uniprot.org/uniprotkb/Q9NXC5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MIOS","classification":"Not Classified","n_dependent_lines":608,"n_total_lines":1208,"dependency_fraction":0.5033112582781457},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"HSP90B1","stoichiometry":0.2},{"gene":"SEC13","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MIOS","total_profiled":1310},"omim":[{"mim_id":"620307","title":"WD REPEAT-CONTAINING PROTEIN 24; WDR24","url":"https://www.omim.org/entry/620307"},{"mim_id":"617418","title":"WD REPEAT-CONTAINING PROTEIN 59; WDR59","url":"https://www.omim.org/entry/617418"},{"mim_id":"617034","title":"CELLULAR ARGININE SENSOR FOR MTORC1 PROTEIN 1; CASTOR1","url":"https://www.omim.org/entry/617034"},{"mim_id":"617033","title":"CELLULAR ARGININE SENSOR FOR MTORC1 PROTEIN 2; CASTOR2","url":"https://www.omim.org/entry/617033"},{"mim_id":"615359","title":"MEIOSIS REGULATOR FOR OOCYTE DEVELOPMENT; MIOS","url":"https://www.omim.org/entry/615359"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":81.5}],"url":"https://www.proteinatlas.org/search/MIOS"},"hgnc":{"alias_symbol":["FLJ20323","MIO","Sea4","Yulink"],"prev_symbol":[]},"alphafold":{"accession":"Q9NXC5","domains":[{"cath_id":"2.130.10.10","chopping":"1-152_170-223","consensus_level":"medium","plddt":89.1445,"start":1,"end":223},{"cath_id":"-","chopping":"819-875","consensus_level":"medium","plddt":81.274,"start":819,"end":875}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NXC5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NXC5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NXC5-F1-predicted_aligned_error_v6.png","plddt_mean":82.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MIOS","jax_strain_url":"https://www.jax.org/strain/search?query=MIOS"},"sequence":{"accession":"Q9NXC5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NXC5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NXC5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NXC5"}},"corpus_meta":[{"pmid":"36220894","id":"PMC_36220894","title":"MiOS, an integrated imaging and computational strategy to model gene folding with nucleosome resolution.","date":"2022","source":"Nature structural & molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/36220894","citation_count":25,"is_preprint":false},{"pmid":"38898112","id":"PMC_38898112","title":"Rbpms2 promotes female fate upstream of the nutrient sensing Gator2 complex component Mios.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38898112","citation_count":12,"is_preprint":false},{"pmid":"38057829","id":"PMC_38057829","title":"The novel roles of YULINK in the migration, proliferation and glycolysis of pulmonary arterial smooth muscle cells: implications for pulmonary arterial hypertension.","date":"2023","source":"Biological research","url":"https://pubmed.ncbi.nlm.nih.gov/38057829","citation_count":7,"is_preprint":false},{"pmid":"33423678","id":"PMC_33423678","title":"Yulink, predicted from evolutionary analysis, is involved in cardiac function.","date":"2021","source":"Journal of biomedical science","url":"https://pubmed.ncbi.nlm.nih.gov/33423678","citation_count":5,"is_preprint":false},{"pmid":"39122768","id":"PMC_39122768","title":"Reverse genetic approaches allowing the characterization of the rabies virus street strain belonging to the SEA4 subclade.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39122768","citation_count":3,"is_preprint":false},{"pmid":"38928292","id":"PMC_38928292","title":"Developing a Tanshinone IIA Memetic by Targeting MIOS to Regulate mTORC1 and Autophagy in Glioblastoma.","date":"2024","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38928292","citation_count":2,"is_preprint":false},{"pmid":"36843032","id":"PMC_36843032","title":"YULINK regulates vascular formation in zebrafish and HUVECs.","date":"2023","source":"Biological research","url":"https://pubmed.ncbi.nlm.nih.gov/36843032","citation_count":1,"is_preprint":false},{"pmid":"41013205","id":"PMC_41013205","title":"YULINK deficiency promotes cell death under glucose restriction in HCC cells in association with GLUT1-mediated glycolysis.","date":"2025","source":"Molecular medicine (Cambridge, Mass.)","url":"https://pubmed.ncbi.nlm.nih.gov/41013205","citation_count":0,"is_preprint":false},{"pmid":"29401584","id":"PMC_29401584","title":"RETRACTED: Deficiency of a novel gene, Yulink, predisposes to heart failure and ventricular arrhythmia.","date":"2018","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/29401584","citation_count":0,"is_preprint":false},{"pmid":"38328218","id":"PMC_38328218","title":"Rbpms2 promotes female fate upstream of the nutrient sensing Gator2 complex component, Mios.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/38328218","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7370,"output_tokens":2091,"usd":0.026738,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9112,"output_tokens":2659,"usd":0.056017,"stage2_stop_reason":"end_turn"},"total_usd":0.082755,"stage1_batch_id":"msgbatch_01BbTCMyrRPyhtEq6b1MdwCN","stage2_batch_id":"msgbatch_011dEed5bbxYvTAaQWKJevxu","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2024,\n      \"finding\": \"Rbpms2 acts upstream of the GATOR2 complex component Mios (MIOS) to promote oocyte fate in zebrafish; genetic analyses indicate Rbpms2 promotes nucleolar amplification via the mTORC1 signaling pathway specifically through Mios, placing Mios as an essential GATOR2-mediated checkpoint component in oogenesis.\",\n      \"method\": \"Genetic epistasis analysis in zebrafish; translational regulation assays; loss-of-function\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with defined pathway placement in zebrafish, single lab, multiple genetic tools but no in vitro reconstitution\",\n      \"pmids\": [\"38898112\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Rbpms2 acts upstream of Mios (MIOS/GATOR2) to promote mTORC1 activation and nucleolar amplification during oogenesis in zebrafish, consistent with Mios functioning as a GATOR2 component that activates mTORC1 signaling in a nutrient-sensing pathway context.\",\n      \"method\": \"Genetic epistasis in zebrafish; loss-of-function\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 2 / Weak — preprint, single lab, overlapping with published version; included as corroboration\",\n      \"pmids\": [\"38328218\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"MIOS (GATOR2 component) is required for Tanshinone IIA (T2A)-mediated inhibition of mTORC1 and induction of autophagy; in Dictyostelium discoideum, ablation of mios abolished T2A-induced autophagy and mTORC1 inhibition, placing MIOS downstream of sestrin 2 (SESN) in the SESN–GATOR2–mTORC1 axis.\",\n      \"method\": \"Genetic ablation (mios- cells) in Dictyostelium discoideum; pharmacological treatment; mTORC1 activity assay; autophagy assay\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean genetic loss-of-function with specific cellular readouts (mTORC1 activity, autophagy), single lab, two orthogonal readouts\",\n      \"pmids\": [\"38928292\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"YULINK (MIOS) knockdown in mouse HL-1 cardiomyocytes reduced DNA binding activity of PPARγ and decreased SERCA2 expression; PPARγ agonists rescued SERCA2 expression in YULINK-KD cells and PPARγ shRNA knockdown phenocopied YULINK loss, indicating YULINK regulates SERCA2 expression through PPARγ nuclear entry.\",\n      \"method\": \"shRNA knockdown in mouse HL-1 cardiomyocytes; DNA binding assay for PPARγ; western blot; agonist rescue; Ca2+ imaging; morpholino knockdown in zebrafish\",\n      \"journal\": \"Journal of biomedical science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (KD, agonist rescue, PPARγ-shRNA phenocopy, DNA binding assay), single lab\",\n      \"pmids\": [\"33423678\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"YULINK (MIOS) co-localizes and physically interacts with endosome-related proteins EPS15, RAB33B, TICAM2 and endocytic markers (Clathrin, RHOB) in HUVECs; YULINK knockdown compromised VEGF-induced VEGFR2 internalization, indicating YULINK participates in endocytic vesicle trafficking to regulate vascular formation.\",\n      \"method\": \"Yeast two-hybrid; FLIM-FRET; immunoprecipitation; immunofluorescence imaging; gene knockdown in zebrafish and HUVECs\",\n      \"journal\": \"Biological research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal binding confirmation by yeast two-hybrid and FLIM-FRET plus IP, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"36843032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"YULINK (MIOS) knockdown in PASMCs suppressed PDGFR expression and phosphorylation of FAK, PI3K, and AKT, indicating YULINK acts upstream of the PI3K-AKT signaling pathway to regulate PASMC migration, proliferation, and glycolysis.\",\n      \"method\": \"shRNA knockdown and overexpression in PASMCs; western blot for pathway components; glucose uptake assay; migration assay; immunofluorescence co-localization\",\n      \"journal\": \"Biological research\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway placement inferred from inhibitor/KD readouts without direct binding evidence for PI3K-AKT components\",\n      \"pmids\": [\"38057829\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"YULINK (MIOS) co-localizes and physically interacts with glucose transporter GLUT1; YULINK knockdown suppressed GLUT1 expression and disrupted GLUT1 translocation from cytosol to cell membrane, resulting in reduced glucose uptake and glycolysis in HCC (Huh7) cells.\",\n      \"method\": \"Proximity ligation assay; immunoprecipitation; immunofluorescence; glucose uptake assay; glycolytic function assay; western blot; shRNA knockdown\",\n      \"journal\": \"Molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct physical interaction confirmed by two orthogonal methods (PLA + IP), functional consequence demonstrated, single lab\",\n      \"pmids\": [\"41013205\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"YULINK (MIOS) regulates SERCA2 expression through PPARγ in cardiomyocytes; YULINK knockdown reduced PPARγ activity and SERCA2 expression, leading to disrupted Ca2+ reuptake and cardiac arrhythmia phenotypes in zebrafish and mouse cardiomyocytes. NOTE: This paper was retracted.\",\n      \"method\": \"Morpholino knockdown in zebrafish; shRNA knockdown in mouse cardiomyocytes; PPARγ activity assay; Ca2+ imaging; western blot [RETRACTED paper]\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 2 / Weak — paper was retracted; findings partially corroborated by later non-retracted work (PMID:33423678) but original report cannot be trusted independently\",\n      \"pmids\": [\"29401584\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MIOS (YULINK/Sea4) is a WD40-repeat-containing subunit of the GATOR2 complex that promotes mTORC1 activation in nutrient-sensing pathways; it physically interacts with GLUT1 to regulate glucose transporter membrane localization and glycolysis, associates with endocytic machinery (EPS15, RAB33B, Clathrin) to support VEGFR2 internalization and vascular formation, and in cardiomyocytes regulates SERCA2 expression via a PPARγ-dependent mechanism to maintain Ca2+ cycling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"MIOS (YULINK/Sea4) is a WD40-repeat subunit of the GATOR2 complex that promotes mTORC1 activation within nutrient-sensing pathways [#0, #2]. In the SESN\\u2013GATOR2\\u2013mTORC1 axis, MIOS acts downstream of sestrin 2 and is required for inhibition of mTORC1 and induction of autophagy, since its ablation in Dictyostelium abolished both responses [#2]; in zebrafish oogenesis it functions as an essential GATOR2-mediated checkpoint downstream of Rbpms2 to drive mTORC1-dependent nucleolar amplification and oocyte fate [#0]. Beyond this canonical role, MIOS engages glucose-handling and trafficking machinery: it physically interacts with the glucose transporter GLUT1 and is required for GLUT1 expression and translocation from cytosol to the plasma membrane, thereby supporting glucose uptake and glycolysis [#6], and it associates with endocytic proteins EPS15, RAB33B, TICAM2, Clathrin and RHOB to support VEGF-induced VEGFR2 internalization and vascular formation [#4]. In cardiomyocytes, MIOS regulates SERCA2 expression through a PPAR\\u03b3-dependent mechanism, controlling Ca2+ reuptake [#3].\",\n  \"teleology\": [\n    {\n      \"year\": 2018,\n      \"claim\": \"An initial question was whether MIOS/YULINK has a role in cardiac calcium handling; loss-of-function linked it to SERCA2 expression and Ca2+ cycling, but the founding report was later retracted.\",\n      \"evidence\": \"Morpholino knockdown in zebrafish and shRNA in mouse cardiomyocytes with PPAR\\u03b3 activity and Ca2+ imaging (retracted)\",\n      \"pmids\": [\"29401584\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Original report retracted; cannot be trusted independently\",\n        \"Mechanism connecting MIOS to PPAR\\u03b3 not defined at the molecular level\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Re-establishing the cardiac role with non-retracted data clarified that MIOS controls SERCA2 expression specifically through promoting PPAR\\u03b3 nuclear/DNA-binding activity.\",\n      \"evidence\": \"shRNA knockdown in HL-1 cardiomyocytes with PPAR\\u03b3 DNA-binding assay, agonist rescue, PPAR\\u03b3-shRNA phenocopy, and Ca2+ imaging\",\n      \"pmids\": [\"33423678\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"How MIOS regulates PPAR\\u03b3 activity mechanistically is unresolved\",\n        \"No demonstrated direct physical interaction between MIOS and PPAR\\u03b3\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Whether MIOS participates in membrane trafficking was tested in endothelial cells, showing it physically associates with endocytic machinery to enable receptor internalization.\",\n      \"evidence\": \"Yeast two-hybrid, FLIM-FRET, IP and immunofluorescence in HUVECs, with VEGFR2 internalization assays and zebrafish knockdown\",\n      \"pmids\": [\"36843032\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct vs. scaffolding role within the endocytic complex not resolved\",\n        \"Relationship between this trafficking function and GATOR2/mTORC1 role unknown\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"A parallel study placed MIOS upstream of PI3K-AKT signaling in vascular smooth muscle, linking it to migration, proliferation and glycolysis.\",\n      \"evidence\": \"shRNA knockdown and overexpression in PASMCs with western blot of pathway components, glucose uptake and migration assays\",\n      \"pmids\": [\"38057829\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Pathway placement inferred from KD readouts without direct binding evidence for PI3K-AKT components\",\n        \"Mechanism connecting MIOS to PDGFR expression not established\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"The canonical GATOR2/mTORC1 function was established in two systems: MIOS is required downstream of sestrin 2 for mTORC1 inhibition and autophagy, and acts as an mTORC1-activating GATOR2 checkpoint in oocyte fate determination.\",\n      \"evidence\": \"Genetic ablation in Dictyostelium with mTORC1 and autophagy readouts; genetic epistasis with Rbpms2 in zebrafish oogenesis (plus corroborating preprint)\",\n      \"pmids\": [\"38928292\", \"38898112\", \"38328218\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Structural basis of MIOS within GATOR2 in these systems not addressed\",\n        \"Whether nutrient-sensing and trafficking/cardiac roles are mechanistically connected is unknown\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Whether MIOS directly controls a glucose transporter was answered by showing physical interaction with GLUT1 and a requirement for GLUT1 membrane translocation, tying MIOS to glycolytic metabolism.\",\n      \"evidence\": \"Proximity ligation assay and IP for interaction; immunofluorescence, glucose uptake and glycolytic assays in Huh7 HCC cells\",\n      \"pmids\": [\"41013205\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which MIOS drives GLUT1 trafficking not defined\",\n        \"Relationship to MIOS endocytic/GATOR2 functions unresolved\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how MIOS's GATOR2/mTORC1 nutrient-sensing function relates mechanistically to its reported roles in GLUT1 trafficking, endocytic VEGFR2 internalization, and PPAR\\u03b3-dependent SERCA2 regulation.\",\n      \"evidence\": \"No timeline study integrates these activities into a single mechanism\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model of MIOS in human cells\",\n        \"Whether trafficking and metabolic roles are mTORC1-dependent is untested\",\n        \"Direct binding partners bridging these functions not identified\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-165159\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"complexes\": [\n      \"GATOR2\"\n    ],\n    \"partners\": [\n      \"GLUT1\",\n      \"EPS15\",\n      \"RAB33B\",\n      \"TICAM2\",\n      \"RHOB\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}