{"gene":"MYOF","run_date":"2026-04-29T11:37:56","timeline":{"discoveries":[{"year":2000,"finding":"Myoferlin (MYOF) was identified as a protein highly homologous to dysferlin, associated with the plasma membrane and nuclear membrane in cardiac and skeletal muscle. It contains multiple C2 domains implicated in calcium-mediated membrane fusion events. Myoferlin expression was found to be upregulated at the membrane in mdx mouse skeletal muscle undergoing repeated degeneration/regeneration cycles, suggesting a role in membrane repair or maintenance.","method":"Database searching, antibody-based subcellular localization (plasma membrane and nuclear membrane association), immunostaining of mdx mouse muscle","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization by immunostaining with functional inference from mdx model; single lab, foundational discovery paper","pmids":["10607832"],"is_preprint":false},{"year":2002,"finding":"The C2A domain of myoferlin (and dysferlin) binds phosphatidylserine in a calcium-sensitive manner. Phospholipid binding by C2A was regulated by calcium concentration, and a dysferlin muscular dystrophy point mutation engineered into C2A reduced calcium-sensitive phospholipid binding. Myoferlin is highly expressed in elongated 'prefusion' myoblasts and is decreased in mature myotubes, whereas dysferlin expression is greatest in mature myotubes, indicating distinct temporal roles in muscle development.","method":"In vitro phospholipid binding assay with calcium titration; site-directed mutagenesis of C2A domain; cell culture model of muscle differentiation with immunostaining","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — in vitro binding assay with mutagenesis and calcium titration, combined with cell biological localization data","pmids":["11959863"],"is_preprint":false},{"year":2019,"finding":"A truncating frameshift variant in human MYOF (c.[2576delG; 2575G>C], p.G859QfsTer8) was identified in a patient with combined cardiomyopathy and limb-girdle muscular dystrophy. Functional studies in patient-derived primary skeletal muscle mesenchymal progenitor cells (RNA sequencing and morphological analysis) confirmed loss-of-function effects, and knockdown of MYOF in zebrafish recapitulated the muscle phenotype, establishing MYOF as causative for human muscle disease.","method":"Whole exome sequencing, RNA sequencing of patient primary cells, morphological analysis, zebrafish in vivo knockdown phenotyping","journal":"Frontiers in genetics","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods including in vitro functional studies and in vivo zebrafish model; single lab","pmids":["31297131"],"is_preprint":false},{"year":2020,"finding":"Myoferlin (MYOF) is a type II transmembrane protein with a single C-terminal transmembrane domain involved in extracellular secretion, endocytosis, vesicle trafficking, membrane repair, membrane receptor recycling, and secreted protein efflux. Small molecule inhibitors targeting the MYOF C2D domain selectively inhibit cancer cell proliferation and migration, identifying C2D as a functional domain mediating MYOF's role in membrane transport processes.","method":"Review of published functional studies; small molecule inhibitor targeting C2D domain with cell proliferation and migration assays","journal":"Current topics in medicinal chemistry","confidence":"Low","confidence_rationale":"Tier 3 — review/summary; C2D inhibitor data referenced but primary experimental detail limited in this source","pmids":["32552653"],"is_preprint":false},{"year":2024,"finding":"Myoferlin associates with influenza A virus (IAV) viral ribonucleoprotein (vRNP) complexes in the cytoplasm during late-stage infection and colocalizes with Rab11a-containing vesicles. This role was conserved in vRNP trafficking of RSV and Sendai virus (SeV). Myoferlin likely recruits EHD family proteins (involved in endosomal biogenesis) to vRNP trafficking endosomes, indicating myoferlin acts as a host co-factor for Rab11a-dependent viral egress.","method":"Co-immunoprecipitation of myoferlin with vRNPs, colocalization imaging with Rab11a, functional knockdown experiments, conservation tested across multiple RNA viruses","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP and colocalization with functional knockdown, replicated across multiple viruses; preprint, single lab","pmids":["bio_10.1101_2024.07.02.601679"],"is_preprint":true},{"year":2024,"finding":"hnRNPLL binds MYOF pre-mRNA and regulates alternative splicing of MYOF exon 17; hnRNPLL depletion causes exon 17 retention, reducing expression of the short MYOF isoform (MYOFb). Overexpression of hnRNPLL or MYOFb promotes pancreatic cancer cell migration and invasion, while hnRNPLL knockdown inhibits metastasis, establishing MYOFb as the functionally pro-metastatic isoform regulated by hnRNPLL-mediated splicing.","method":"RNA-seq after hnRNPLL knockdown, splicing reporter assays, isoform-specific overexpression and knockdown with migration/invasion assays","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 — isoform-specific functional rescue experiments with multiple cell biology readouts; single lab","pmids":["39742990"],"is_preprint":false},{"year":2025,"finding":"MYOF promotes PDAC progression by stabilizing ILF3 protein through recruiting OTUB1 deubiquitinase: MYOF physically recruits both OTUB1 and ILF3, enhancing their interaction and relieving K48-linked ubiquitination and proteasomal degradation of ILF3. Stabilized ILF3 in turn binds and stabilizes LCN2 mRNA, increasing LCN2 expression to suppress ferroptosis. MYOF knockout promotes ferroptosis via LCN2 downregulation, and ectopic LCN2 rescues the MYOF-KO phenotype.","method":"Co-immunoprecipitation of MYOF-OTUB1-ILF3 complex, ubiquitination assays, mRNA stability assays, MYOF knockout in vitro and in vivo xenograft, genetic epistasis (LCN2 rescue)","journal":"Redox biology","confidence":"High","confidence_rationale":"Tier 1-2 — reciprocal Co-IP, ubiquitination assays, mRNA stability, genetic epistasis rescue, and in vivo validation with multiple orthogonal methods","pmids":["40381229"],"is_preprint":false},{"year":2025,"finding":"TRIM8, an E3 ubiquitin ligase, targets MYOF for K48-linked polyubiquitination, leading to proteasome-mediated degradation of MYOF. TRIM8 overexpression reduced MYOF protein levels in NSCLC cells; restoring MYOF expression rescued cell migration in TRIM8-overexpressing cells. MYOF degradation by TRIM8 subsequently suppressed extracellular secretion of matrix metalloproteinases (MMPs), inhibiting lung cancer metastasis.","method":"Co-immunoprecipitation, K48-linked ubiquitination assays, gain- and loss-of-function experiments, MYOF rescue in TRIM8-overexpressing cells, xenograft in vivo model, MMP secretion assays","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 1-2 — direct ubiquitination assay with linkage-specific characterization, reciprocal epistasis rescue, and in vivo validation; single lab but multiple orthogonal methods","pmids":["39934162"],"is_preprint":false}],"current_model":"MYOF (myoferlin) is a multi-C2-domain transmembrane protein whose C2A domain binds phosphatidylserine in a calcium-sensitive manner to mediate membrane repair and fusion; it is regulated post-translationally by TRIM8-mediated K48-linked ubiquitination and proteasomal degradation, and it promotes cancer progression by recruiting OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA to suppress ferroptosis, while also facilitating viral egress by associating with Rab11a/vRNP vesicles and recruiting EHD proteins."},"narrative":{"teleology":[{"year":2000,"claim":"Identification of MYOF as a dysferlin-homologous, multi-C2-domain protein at the plasma and nuclear membranes of muscle cells established a new candidate for calcium-dependent membrane fusion in striated muscle.","evidence":"Database homology search and immunostaining in cardiac/skeletal muscle including mdx mouse","pmids":["10607832"],"confidence":"Medium","gaps":["No direct functional assay for membrane fusion activity","Expression pattern characterized only in muscle; non-muscle roles unknown"]},{"year":2002,"claim":"Demonstrating that the C2A domain binds phosphatidylserine in a calcium-titratable fashion, and that a disease-relevant mutation disrupts this binding, provided the first mechanistic link between MYOF's C2 domains and lipid-membrane interaction required for its fusogenic role.","evidence":"In vitro phospholipid binding assay with calcium titration and site-directed mutagenesis of C2A domain","pmids":["11959863"],"confidence":"High","gaps":["Only C2A tested; functional contributions of other C2 domains (C2B–C2F) uncharacterized","No in vivo membrane fusion assay performed"]},{"year":2019,"claim":"Discovery of a truncating MYOF variant in a patient with cardiomyopathy and limb-girdle muscular dystrophy, supported by zebrafish phenocopy, established MYOF as a bona fide human disease gene for combined skeletal–cardiac myopathy.","evidence":"Whole exome sequencing, RNA-seq of patient primary cells, zebrafish knockdown phenotyping","pmids":["31297131"],"confidence":"Medium","gaps":["Single family reported; genetic spectrum and prevalence unknown","Precise cellular mechanism of muscle degeneration not delineated"]},{"year":2024,"claim":"Identification of hnRNPLL-regulated alternative splicing of MYOF exon 17 revealed that the short isoform MYOFb is the functionally pro-metastatic species in pancreatic cancer, adding an RNA-level regulatory layer to MYOF biology.","evidence":"RNA-seq after hnRNPLL knockdown, splicing reporters, isoform-specific overexpression/knockdown with migration and invasion assays","pmids":["39742990"],"confidence":"Medium","gaps":["Structural or functional difference between isoforms at the protein level not defined","Relevance of isoform switching in non-cancer contexts unknown"]},{"year":2024,"claim":"Showing that MYOF associates with Rab11a-positive vRNP vesicles during influenza A infection and recruits EHD proteins expanded MYOF's vesicle trafficking role to viral egress, conserved across multiple RNA viruses.","evidence":"Co-immunoprecipitation with vRNPs, Rab11a colocalization imaging, knockdown across IAV/RSV/SeV (preprint)","pmids":["bio_10.1101_2024.07.02.601679"],"confidence":"Medium","gaps":["Preprint; not yet peer-reviewed","Mechanism by which MYOF recruits EHD proteins not resolved at the domain level","Whether MYOF's role in viral egress is linked to its membrane repair function is untested"]},{"year":2025,"claim":"Elucidation of a MYOF–OTUB1–ILF3 axis in pancreatic cancer revealed that MYOF acts as a scaffold recruiting the deubiquitinase OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA and suppresses ferroptosis — providing a direct oncogenic mechanism beyond membrane dynamics.","evidence":"Reciprocal Co-IP, K48-ubiquitination assays, mRNA stability assays, MYOF knockout with LCN2 epistasis rescue, in vivo xenograft","pmids":["40381229"],"confidence":"High","gaps":["Which MYOF domain mediates OTUB1 and ILF3 recruitment is unknown","Whether this scaffolding function operates outside pancreatic cancer is untested"]},{"year":2025,"claim":"Demonstration that TRIM8 targets MYOF for K48-linked ubiquitination and proteasomal degradation, thereby suppressing MMP secretion and metastasis, established the first defined E3 ligase-mediated regulatory mechanism controlling MYOF protein turnover.","evidence":"K48-linkage-specific ubiquitination assays, TRIM8 gain/loss-of-function, MYOF rescue in TRIM8-overexpressing NSCLC cells, xenograft model, MMP secretion assays","pmids":["39934162"],"confidence":"High","gaps":["TRIM8 recognition motif on MYOF not mapped","Whether TRIM8 regulation of MYOF operates in muscle or non-cancer contexts is unknown"]},{"year":null,"claim":"The structural basis for MYOF's multi-C2-domain architecture in membrane fusion, the precise domain requirements for its scaffolding of OTUB1/ILF3 and EHD recruitment, and whether its muscle and cancer functions share a common vesicle-trafficking mechanism remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution structure of full-length MYOF or individual C2 domains beyond C2A","Functional contributions of C2B–C2F domains are largely uncharacterized","Integration of membrane repair, vesicle trafficking, and scaffolding functions into a unified model is lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[1]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[6]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,1]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[4]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[4]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[4,7]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[6,7]}],"complexes":[],"partners":["OTUB1","ILF3","TRIM8","RAB11A","HNRNPLL"],"other_free_text":[]},"mechanistic_narrative":"Myoferlin (MYOF) is a multi-C2-domain, type II transmembrane protein that mediates calcium-dependent membrane fusion, vesicle trafficking, and membrane repair, with particular importance in muscle and in cancer cell membrane dynamics. Its C2A domain binds phosphatidylserine in a calcium-sensitive manner, and MYOF is expressed most highly in prefusion myoblasts, distinguishing it temporally from the related protein dysferlin [PMID:11959863]. Loss-of-function mutations in MYOF cause combined cardiomyopathy and limb-girdle muscular dystrophy in humans, confirmed by zebrafish phenocopy [PMID:31297131]. In pancreatic and lung cancers, MYOF promotes progression by recruiting the deubiquitinase OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA and suppresses ferroptosis [PMID:40381229], while MYOF protein levels are themselves controlled by TRIM8-mediated K48-linked ubiquitination and proteasomal degradation, which limits MMP secretion and metastasis [PMID:39934162]."},"prefetch_data":{"uniprot":{"accession":"Q9NZM1","full_name":"Myoferlin","aliases":["Fer-1-like protein 3"],"length_aa":2061,"mass_kda":234.7,"function":"Calcium/phospholipid-binding protein that plays a role in the plasmalemma repair mechanism of endothelial cells that permits rapid resealing of membranes disrupted by mechanical stress. Involved in endocytic recycling. Implicated in VEGF signal transduction by regulating the levels of the receptor KDR (By similarity)","subcellular_location":"Cell membrane; Nucleus membrane; Cytoplasmic vesicle membrane; Late endosome membrane","url":"https://www.uniprot.org/uniprotkb/Q9NZM1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/MYOF","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/MYOF","total_profiled":1310},"omim":[{"mim_id":"619366","title":"ANGIOEDEMA, HEREDITARY, 7; HAE7","url":"https://www.omim.org/entry/619366"},{"mim_id":"604603","title":"MYOFERLIN; MYOF","url":"https://www.omim.org/entry/604603"},{"mim_id":"603009","title":"DYSFERLIN; DYSF","url":"https://www.omim.org/entry/603009"},{"mim_id":"106100","title":"ANGIOEDEMA, HEREDITARY, 1; HAE1","url":"https://www.omim.org/entry/106100"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Vesicles","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Supported"},{"location":"Centriolar satellite","reliability":"Supported"},{"location":"Primary cilium","reliability":"Additional"},{"location":"Primary cilium tip","reliability":"Additional"},{"location":"Basal body","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MYOF"},"hgnc":{"alias_symbol":["KIAA1207"],"prev_symbol":["FER1L3"]},"alphafold":{"accession":"Q9NZM1","domains":[{"cath_id":"2.60.40.150","chopping":"3-125","consensus_level":"high","plddt":87.5181,"start":3,"end":125},{"cath_id":"2.60.40.150","chopping":"198-331_339-354","consensus_level":"high","plddt":80.9095,"start":198,"end":354},{"cath_id":"2.60.40.150","chopping":"356-469_495-570","consensus_level":"medium","plddt":78.8629,"start":356,"end":570},{"cath_id":"-","chopping":"931-962_1017-1029","consensus_level":"medium","plddt":85.0922,"start":931,"end":1029},{"cath_id":"2.60.40.150","chopping":"1136-1299","consensus_level":"medium","plddt":86.2299,"start":1136,"end":1299},{"cath_id":"2.60.40.150","chopping":"1300-1417_1492-1534","consensus_level":"medium","plddt":81.8071,"start":1300,"end":1534},{"cath_id":"2.60.40.150","chopping":"1552-1653_1742-1773","consensus_level":"medium","plddt":87.8244,"start":1552,"end":1773},{"cath_id":"2.60.40.150","chopping":"1787-1928_1935-2006","consensus_level":"high","plddt":83.791,"start":1787,"end":2006},{"cath_id":"1.20.120","chopping":"652-714_718-766","consensus_level":"medium","plddt":87.0914,"start":652,"end":766},{"cath_id":"1.20.5","chopping":"2009-2049","consensus_level":"medium","plddt":69.439,"start":2009,"end":2049}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZM1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZM1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NZM1-F1-predicted_aligned_error_v6.png","plddt_mean":79.38},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MYOF","jax_strain_url":"https://www.jax.org/strain/search?query=MYOF"},"sequence":{"accession":"Q9NZM1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NZM1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NZM1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NZM1"}},"corpus_meta":[{"pmid":"12953059","id":"PMC_12953059","title":"Shared, 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Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11959863","citation_count":168,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"30711629","id":"PMC_30711629","title":"MET Inhibitors Promote Liver Tumor Evasion of the Immune Response by Stabilizing PDL1.","date":"2019","source":"Gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/30711629","citation_count":165,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19913121","id":"PMC_19913121","title":"Gene-centric association signals for lipids and apolipoproteins identified via the HumanCVD BeadChip.","date":"2009","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/19913121","citation_count":164,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"30833792","id":"PMC_30833792","title":"A protein-interaction network of interferon-stimulated genes extends the innate immune system landscape.","date":"2019","source":"Nature immunology","url":"https://pubmed.ncbi.nlm.nih.gov/30833792","citation_count":159,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"10607832","id":"PMC_10607832","title":"Myoferlin, a candidate gene and potential modifier of muscular dystrophy.","date":"2000","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/10607832","citation_count":153,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"23533145","id":"PMC_23533145","title":"In-depth proteomic analyses of exosomes isolated from expressed prostatic secretions in urine.","date":"2013","source":"Proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/23533145","citation_count":138,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"16385451","id":"PMC_16385451","title":"A scan of chromosome 10 identifies a novel locus showing strong association with late-onset Alzheimer disease.","date":"2005","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/16385451","citation_count":137,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"31871319","id":"PMC_31871319","title":"Mapping the proximity interaction network of the Rho-family GTPases reveals signalling pathways and regulatory mechanisms.","date":"2019","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/31871319","citation_count":137,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":7887,"output_tokens":1324,"usd":0.02176},"stage2":{"model":"claude-opus-4-6","input_tokens":4540,"output_tokens":1756,"usd":0.0999},"total_usd":0.306299,"stage1_batch_id":"msgbatch_012ZiiCUBfB62NCTQd4Bk1Jo","stage2_batch_id":"msgbatch_01Xuqq9aUHoQouS7UVGZNStZ","note":"batch pricing = 50% of 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(patient cells + zebrafish), single lab\",\n      \"pmids\": [\"31297131\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM8, an E3 ubiquitin ligase, directly targets MYOF for K48-linked polyubiquitination and proteasome-mediated degradation; TRIM8 overexpression reduces MYOF protein levels, suppresses extracellular MMP secretion, and inhibits NSCLC cell migration/invasion, whereas restoring MYOF rescues migration in TRIM8-overexpressing cells.\",\n      \"method\": \"Co-IP, ubiquitination assay (K48 linkage specificity), gain- and loss-of-function experiments, xenograft model, MYOF rescue\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal functional rescue and K48-linkage specificity assay, single lab\",\n      \"pmids\": [\"39934162\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MYOF recruits both OTUB1 (a deubiquitinase) and ILF3 (an RNA-binding protein) and enhances their interaction, thereby stabilizing ILF3 protein by relieving its ubiquitination and proteasomal degradation; stabilized ILF3 then binds LCN2 mRNA to increase its stability, promoting PDAC progression and suppressing ferroptosis.\",\n      \"method\": \"Co-IP, knockout rescue, mRNA stability assay, in vitro and in vivo PDAC models\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple Co-IPs, KO rescue with ectopic LCN2, in vivo validation, single lab\",\n      \"pmids\": [\"40381229\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"hnRNPLL regulates alternative splicing of MYOF; its depletion causes MYOF exon 17 retention, reducing the short isoform MYOFb and inhibiting pancreatic cancer cell migration and invasion; overexpression of hnRNPLL or MYOFb promotes migration and invasion.\",\n      \"method\": \"RNA splicing assays, knockdown/overexpression, migration/invasion assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — gain- and loss-of-function with defined splicing mechanism, single lab\",\n      \"pmids\": [\"39742990\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Myoferlin associates with influenza A virus (IAV) vRNP complexes in the cytoplasm and colocalizes with Rab11a-containing vesicles during late-stage infection; this role is conserved for RSV and SeV vRNP trafficking; myoferlin likely recruits EHD family proteins involved in endosomal biogenesis to these vRNP trafficking endosomes.\",\n      \"method\": \"Co-localization imaging, Co-IP/pulldown, multi-virus functional assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — co-localization and association data across multiple viruses, single lab, preprint\",\n      \"pmids\": [\"bio_10.1101_2024.07.02.601679\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"MYOF (myoferlin) is a ferlin-family transmembrane protein that functions in membrane-associated processes including vesicle trafficking, endosomal recycling, and membrane repair; it acts as a scaffold that recruits the deubiquitinase OTUB1 and RNA-binding protein ILF3 to stabilize downstream signaling (LCN2 mRNA), is subject to K48-linked ubiquitination and proteasomal degradation mediated by the E3 ligase TRIM8, undergoes hnRNPLL-regulated alternative splicing that generates a pro-migratory short isoform, and is required for normal skeletal muscle integrity; it also associates with Rab11a-positive endosomal vesicles to facilitate viral vRNP egress.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\nI'll classify each paper first, then extract discoveries only from KEEP papers.\n\n**Classification:**\n\nRetrieved papers:\n1. PMID:12953059 — Dictyostelium MyoF (class I myosin) — EXCLUDE: Symbol collision; Dictyostelium MyoF is a class I myosin, fundamentally different protein from human MYOF (ferlin family)\n2. PMID:7787058 — Dictyostelium myoF — EXCLUDE: Same as above\n3. PMID:34464754 — Trypanosoma cruzi MyoF (orphan myosin) — EXCLUDE: Symbol collision; unrelated myosin motor protein\n4. PMID:31297131 — Human MYOF (myoferlin), muscular dystrophy — KEEP\n5. PMID:32552653 — Human MYOF (myoferlin) review — KEEP\n6. PMID:39742990 — Human MYOF (myoferlin), splicing — KEEP\n7. PMID:40381229 — Human MYOF (myoferlin), PDAC — KEEP\n8. PMID:39934162 — Human MYOF (myoferlin), TRIM8 — KEEP\n9. PMID:38253296 — Human MYOF (myoferlin), ARVC — KEEP (genetic association, but limited mechanism)\n10. PMID:38054026 — Human MYOF (myoferlin), SCLS — KEEP (but minimal mechanism)\n11. PMID:40665025 — in silico/bioinformatics only — EXCLUDE (Tier 4 only)\n12. PMID:39498317 — Corrigendum — EXCLUDE\n13. bio_10.1101_2025.10.02.679852 — Toxoplasma MyoF (myosin motor) — EXCLUDE: Symbol collision\n14. bio_10.1101_2024.07.02.601679 — Human myoferlin (MYOF), viral trafficking — KEEP\n15. bio_10.1101_2025.08.15.670481 — Toxoplasma MyoF — EXCLUDE: Symbol collision\n16. bio_10.1101_2024.10.18.619080 — Integrin/ADPKD study, no MYOF focus — EXCLUDE\n\nGene2pubmed curated papers:\n- PMID:17081983 — phosphoproteomics, MYOF detected incidentally — EXCLUDE (no mechanistic finding about MYOF)\n- PMID:12477932 — cDNA sequencing — EXCLUDE\n- PMID:15302935 — phosphoproteomics — EXCLUDE\n- PMID:26496610 — interactome — EXCLUDE (no specific MYOF mechanism)\n- PMID:29507755 — m6A methylation, no MYOF focus — EXCLUDE\n- PMID:18854154 — HIV host factors — EXCLUDE\n- PMID:11790298 — nucleolar proteomics — EXCLUDE\n- PMID:14702039 — cDNA sequencing — EXCLUDE\n- PMID:33961781 — interactome — EXCLUDE\n- PMID:22939629 — protein complexes — EXCLUDE\n- PMID:21873635 — GO annotation — EXCLUDE\n- PMID:19056867 — urinary exosomes proteomics — EXCLUDE\n- PMID:33845483 — SARS-CoV-2 proteomics — EXCLUDE\n- PMID:15489334 — cDNA — EXCLUDE\n- PMID:16009940 — ISG15 targets — EXCLUDE\n- PMID:34079125 — BioID map — EXCLUDE\n- PMID:28388416 — APEX proximity labeling — EXCLUDE\n- PMID:22810586 — tumor virus interactome — EXCLUDE\n- PMID:27342126 — Zika/Dengue genomics — EXCLUDE\n- PMID:36369321 — CST1/OTUB1/GPX4 in gastric cancer — EXCLUDE (about CST1, not MYOF)\n- PMID:24332808 — PRP19 — EXCLUDE\n- PMID:29568061 — MAC-tag proteomics — EXCLUDE\n- PMID:11959863 — Davis et al., ferlin C2 domains — KEEP (directly about myoferlin C2 domains)\n- PMID:30711629 — MET/PDL1 — EXCLUDE\n- PMID:19913121 — lipid GWAS — EXCLUDE\n- PMID:30833792 — ISG network — EXCLUDE\n- PMID:10607832 — Davis et al. 2000, myoferlin discovery — KEEP\n- PMID:23533145 — EPS exosomes proteomics — EXCLUDE\n- PMID:16385451 — Alzheimer's GWAS — EXCLUDE\n- PMID:31871319 — Rho GTPase network — EXCLUDE\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2000,\n      \"finding\": \"Myoferlin (MYOF) was identified as a protein highly homologous to dysferlin, associated with the plasma membrane and nuclear membrane in cardiac and skeletal muscle. It contains multiple C2 domains implicated in calcium-mediated membrane fusion events. Myoferlin expression was found to be upregulated at the membrane in mdx mouse skeletal muscle undergoing repeated degeneration/regeneration cycles, suggesting a role in membrane repair or maintenance.\",\n      \"method\": \"Database searching, antibody-based subcellular localization (plasma membrane and nuclear membrane association), immunostaining of mdx mouse muscle\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization by immunostaining with functional inference from mdx model; single lab, foundational discovery paper\",\n      \"pmids\": [\"10607832\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"The C2A domain of myoferlin (and dysferlin) binds phosphatidylserine in a calcium-sensitive manner. Phospholipid binding by C2A was regulated by calcium concentration, and a dysferlin muscular dystrophy point mutation engineered into C2A reduced calcium-sensitive phospholipid binding. Myoferlin is highly expressed in elongated 'prefusion' myoblasts and is decreased in mature myotubes, whereas dysferlin expression is greatest in mature myotubes, indicating distinct temporal roles in muscle development.\",\n      \"method\": \"In vitro phospholipid binding assay with calcium titration; site-directed mutagenesis of C2A domain; cell culture model of muscle differentiation with immunostaining\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro binding assay with mutagenesis and calcium titration, combined with cell biological localization data\",\n      \"pmids\": [\"11959863\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"A truncating frameshift variant in human MYOF (c.[2576delG; 2575G>C], p.G859QfsTer8) was identified in a patient with combined cardiomyopathy and limb-girdle muscular dystrophy. Functional studies in patient-derived primary skeletal muscle mesenchymal progenitor cells (RNA sequencing and morphological analysis) confirmed loss-of-function effects, and knockdown of MYOF in zebrafish recapitulated the muscle phenotype, establishing MYOF as causative for human muscle disease.\",\n      \"method\": \"Whole exome sequencing, RNA sequencing of patient primary cells, morphological analysis, zebrafish in vivo knockdown phenotyping\",\n      \"journal\": \"Frontiers in genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods including in vitro functional studies and in vivo zebrafish model; single lab\",\n      \"pmids\": [\"31297131\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Myoferlin (MYOF) is a type II transmembrane protein with a single C-terminal transmembrane domain involved in extracellular secretion, endocytosis, vesicle trafficking, membrane repair, membrane receptor recycling, and secreted protein efflux. Small molecule inhibitors targeting the MYOF C2D domain selectively inhibit cancer cell proliferation and migration, identifying C2D as a functional domain mediating MYOF's role in membrane transport processes.\",\n      \"method\": \"Review of published functional studies; small molecule inhibitor targeting C2D domain with cell proliferation and migration assays\",\n      \"journal\": \"Current topics in medicinal chemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — review/summary; C2D inhibitor data referenced but primary experimental detail limited in this source\",\n      \"pmids\": [\"32552653\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Myoferlin associates with influenza A virus (IAV) viral ribonucleoprotein (vRNP) complexes in the cytoplasm during late-stage infection and colocalizes with Rab11a-containing vesicles. This role was conserved in vRNP trafficking of RSV and Sendai virus (SeV). Myoferlin likely recruits EHD family proteins (involved in endosomal biogenesis) to vRNP trafficking endosomes, indicating myoferlin acts as a host co-factor for Rab11a-dependent viral egress.\",\n      \"method\": \"Co-immunoprecipitation of myoferlin with vRNPs, colocalization imaging with Rab11a, functional knockdown experiments, conservation tested across multiple RNA viruses\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and colocalization with functional knockdown, replicated across multiple viruses; preprint, single lab\",\n      \"pmids\": [\"bio_10.1101_2024.07.02.601679\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"hnRNPLL binds MYOF pre-mRNA and regulates alternative splicing of MYOF exon 17; hnRNPLL depletion causes exon 17 retention, reducing expression of the short MYOF isoform (MYOFb). Overexpression of hnRNPLL or MYOFb promotes pancreatic cancer cell migration and invasion, while hnRNPLL knockdown inhibits metastasis, establishing MYOFb as the functionally pro-metastatic isoform regulated by hnRNPLL-mediated splicing.\",\n      \"method\": \"RNA-seq after hnRNPLL knockdown, splicing reporter assays, isoform-specific overexpression and knockdown with migration/invasion assays\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — isoform-specific functional rescue experiments with multiple cell biology readouts; single lab\",\n      \"pmids\": [\"39742990\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MYOF promotes PDAC progression by stabilizing ILF3 protein through recruiting OTUB1 deubiquitinase: MYOF physically recruits both OTUB1 and ILF3, enhancing their interaction and relieving K48-linked ubiquitination and proteasomal degradation of ILF3. Stabilized ILF3 in turn binds and stabilizes LCN2 mRNA, increasing LCN2 expression to suppress ferroptosis. MYOF knockout promotes ferroptosis via LCN2 downregulation, and ectopic LCN2 rescues the MYOF-KO phenotype.\",\n      \"method\": \"Co-immunoprecipitation of MYOF-OTUB1-ILF3 complex, ubiquitination assays, mRNA stability assays, MYOF knockout in vitro and in vivo xenograft, genetic epistasis (LCN2 rescue)\",\n      \"journal\": \"Redox biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — reciprocal Co-IP, ubiquitination assays, mRNA stability, genetic epistasis rescue, and in vivo validation with multiple orthogonal methods\",\n      \"pmids\": [\"40381229\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"TRIM8, an E3 ubiquitin ligase, targets MYOF for K48-linked polyubiquitination, leading to proteasome-mediated degradation of MYOF. TRIM8 overexpression reduced MYOF protein levels in NSCLC cells; restoring MYOF expression rescued cell migration in TRIM8-overexpressing cells. MYOF degradation by TRIM8 subsequently suppressed extracellular secretion of matrix metalloproteinases (MMPs), inhibiting lung cancer metastasis.\",\n      \"method\": \"Co-immunoprecipitation, K48-linked ubiquitination assays, gain- and loss-of-function experiments, MYOF rescue in TRIM8-overexpressing cells, xenograft in vivo model, MMP secretion assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct ubiquitination assay with linkage-specific characterization, reciprocal epistasis rescue, and in vivo validation; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"39934162\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MYOF (myoferlin) is a multi-C2-domain transmembrane protein whose C2A domain binds phosphatidylserine in a calcium-sensitive manner to mediate membrane repair and fusion; it is regulated post-translationally by TRIM8-mediated K48-linked ubiquitination and proteasomal degradation, and it promotes cancer progression by recruiting OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA to suppress ferroptosis, while also facilitating viral egress by associating with Rab11a/vRNP vesicles and recruiting EHD proteins.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MYOF (myoferlin) is a ferlin-family membrane protein that functions as a multivalent scaffold integrating vesicle trafficking, membrane dynamics, and post-transcriptional signaling. It recruits the deubiquitinase OTUB1 and the RNA-binding protein ILF3, bridging their interaction to stabilize ILF3 and thereby increase LCN2 mRNA stability, which suppresses ferroptosis and promotes pancreatic cancer progression [PMID:40381229]. MYOF protein levels are regulated by TRIM8-mediated K48-linked polyubiquitination and proteasomal degradation, and its alternative splicing by hnRNPLL generates a short isoform (MYOFb) that promotes cancer cell migration and invasion [PMID:39934162, PMID:39742990]. A truncating MYOF variant causes limb-girdle muscular dystrophy with cardiomyopathy, establishing a requirement for MYOF in skeletal and cardiac muscle integrity [PMID:31297131].\",\n  \"teleology\": [\n    {\n      \"year\": 2019,\n      \"claim\": \"Establishing that MYOF loss causes human disease answered whether this ferlin-family member is essential for muscle integrity, linking it to limb-girdle muscular dystrophy and cardiomyopathy.\",\n      \"evidence\": \"Patient-derived skeletal muscle progenitor cell studies (RNA-seq, morphology) and zebrafish in vivo knockdown recapitulating the muscle phenotype\",\n      \"pmids\": [\"31297131\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single family study; additional pedigrees or rescue experiments in patient cells would strengthen the causal link\",\n        \"Molecular mechanism by which MYOF maintains muscle membrane integrity not defined\",\n        \"Relationship between muscle repair function and vesicle trafficking roles not established\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrating that hnRNPLL-regulated alternative splicing of MYOF generates a pro-migratory short isoform (MYOFb) revealed how isoform-specific functions arise and influence cancer cell invasion.\",\n      \"evidence\": \"RNA splicing assays, knockdown/overexpression of hnRNPLL and MYOFb, migration and invasion assays in pancreatic cancer cells\",\n      \"pmids\": [\"39742990\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Structural or domain-level differences between MYOF isoforms that confer distinct functions not characterized\",\n        \"Whether MYOFb splicing is relevant in non-cancer tissues is unknown\",\n        \"Single lab; independent replication needed\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showing that MYOF associates with Rab11a-positive endosomal vesicles carrying viral vRNP complexes extended its trafficking role to host–pathogen interactions and suggested EHD protein recruitment to these endosomes.\",\n      \"evidence\": \"Co-localization imaging, Co-IP/pulldown, multi-virus functional assays (IAV, RSV, SeV) (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.07.02.601679\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Preprint; not yet peer reviewed\",\n        \"Direct interaction between MYOF and Rab11a or EHD proteins not biochemically validated\",\n        \"Whether MYOF's role in viral egress depends on the same domains as its muscle or cancer functions is unknown\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identifying TRIM8 as the E3 ligase that marks MYOF for K48-linked ubiquitination and proteasomal degradation defined the primary turnover mechanism controlling MYOF protein levels and downstream MMP secretion.\",\n      \"evidence\": \"Co-IP, K48-linkage-specific ubiquitination assay, gain/loss-of-function with MYOF rescue, NSCLC xenograft model\",\n      \"pmids\": [\"39934162\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Specific MYOF lysine residues targeted by TRIM8 not identified\",\n        \"Whether TRIM8-mediated degradation operates in non-cancer contexts (e.g., muscle) is untested\",\n        \"Single lab; independent replication needed\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealing that MYOF scaffolds the OTUB1–ILF3 interaction to stabilize ILF3 and thereby increase LCN2 mRNA stability defined a post-transcriptional signaling axis by which MYOF suppresses ferroptosis and drives PDAC progression.\",\n      \"evidence\": \"Co-IP, knockout rescue with ectopic LCN2, mRNA stability assays, in vitro and in vivo PDAC models\",\n      \"pmids\": [\"40381229\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Domain on MYOF responsible for OTUB1 and ILF3 binding not mapped\",\n        \"Whether this scaffolding function depends on membrane association or occurs in the cytosol is unclear\",\n        \"Single lab; independent validation needed\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How MYOF's diverse roles — membrane scaffolding, vesicle trafficking, mRNA stabilization, and muscle integrity — are coordinated through its multi-C2 domain architecture remains an open question.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural model of full-length MYOF or domain-specific functional mapping exists\",\n        \"Whether the OTUB1/ILF3 scaffolding and Rab11a vesicle trafficking functions are mutually exclusive or co-occurring is unknown\",\n        \"Physiological relevance of TRIM8-mediated degradation outside cancer models not tested\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1]},\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-392499\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"TRIM8\",\n      \"OTUB1\",\n      \"ILF3\",\n      \"HNRNPLL\",\n      \"RAB11A\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"Myoferlin (MYOF) is a multi-C2-domain, type II transmembrane protein that mediates calcium-dependent membrane fusion, vesicle trafficking, and membrane repair, with particular importance in muscle and in cancer cell membrane dynamics. Its C2A domain binds phosphatidylserine in a calcium-sensitive manner, and MYOF is expressed most highly in prefusion myoblasts, distinguishing it temporally from the related protein dysferlin [PMID:11959863]. Loss-of-function mutations in MYOF cause combined cardiomyopathy and limb-girdle muscular dystrophy in humans, confirmed by zebrafish phenocopy [PMID:31297131]. In pancreatic and lung cancers, MYOF promotes progression by recruiting the deubiquitinase OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA and suppresses ferroptosis [PMID:40381229], while MYOF protein levels are themselves controlled by TRIM8-mediated K48-linked ubiquitination and proteasomal degradation, which limits MMP secretion and metastasis [PMID:39934162].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Identification of MYOF as a dysferlin-homologous, multi-C2-domain protein at the plasma and nuclear membranes of muscle cells established a new candidate for calcium-dependent membrane fusion in striated muscle.\",\n      \"evidence\": \"Database homology search and immunostaining in cardiac/skeletal muscle including mdx mouse\",\n      \"pmids\": [\"10607832\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No direct functional assay for membrane fusion activity\",\n        \"Expression pattern characterized only in muscle; non-muscle roles unknown\"\n      ]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Demonstrating that the C2A domain binds phosphatidylserine in a calcium-titratable fashion, and that a disease-relevant mutation disrupts this binding, provided the first mechanistic link between MYOF's C2 domains and lipid-membrane interaction required for its fusogenic role.\",\n      \"evidence\": \"In vitro phospholipid binding assay with calcium titration and site-directed mutagenesis of C2A domain\",\n      \"pmids\": [\"11959863\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Only C2A tested; functional contributions of other C2 domains (C2B–C2F) uncharacterized\",\n        \"No in vivo membrane fusion assay performed\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Discovery of a truncating MYOF variant in a patient with cardiomyopathy and limb-girdle muscular dystrophy, supported by zebrafish phenocopy, established MYOF as a bona fide human disease gene for combined skeletal–cardiac myopathy.\",\n      \"evidence\": \"Whole exome sequencing, RNA-seq of patient primary cells, zebrafish knockdown phenotyping\",\n      \"pmids\": [\"31297131\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Single family reported; genetic spectrum and prevalence unknown\",\n        \"Precise cellular mechanism of muscle degeneration not delineated\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identification of hnRNPLL-regulated alternative splicing of MYOF exon 17 revealed that the short isoform MYOFb is the functionally pro-metastatic species in pancreatic cancer, adding an RNA-level regulatory layer to MYOF biology.\",\n      \"evidence\": \"RNA-seq after hnRNPLL knockdown, splicing reporters, isoform-specific overexpression/knockdown with migration and invasion assays\",\n      \"pmids\": [\"39742990\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Structural or functional difference between isoforms at the protein level not defined\",\n        \"Relevance of isoform switching in non-cancer contexts unknown\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showing that MYOF associates with Rab11a-positive vRNP vesicles during influenza A infection and recruits EHD proteins expanded MYOF's vesicle trafficking role to viral egress, conserved across multiple RNA viruses.\",\n      \"evidence\": \"Co-immunoprecipitation with vRNPs, Rab11a colocalization imaging, knockdown across IAV/RSV/SeV (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.07.02.601679\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Preprint; not yet peer-reviewed\",\n        \"Mechanism by which MYOF recruits EHD proteins not resolved at the domain level\",\n        \"Whether MYOF's role in viral egress is linked to its membrane repair function is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Elucidation of a MYOF–OTUB1–ILF3 axis in pancreatic cancer revealed that MYOF acts as a scaffold recruiting the deubiquitinase OTUB1 to stabilize ILF3, which in turn stabilizes LCN2 mRNA and suppresses ferroptosis — providing a direct oncogenic mechanism beyond membrane dynamics.\",\n      \"evidence\": \"Reciprocal Co-IP, K48-ubiquitination assays, mRNA stability assays, MYOF knockout with LCN2 epistasis rescue, in vivo xenograft\",\n      \"pmids\": [\"40381229\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Which MYOF domain mediates OTUB1 and ILF3 recruitment is unknown\",\n        \"Whether this scaffolding function operates outside pancreatic cancer is untested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstration that TRIM8 targets MYOF for K48-linked ubiquitination and proteasomal degradation, thereby suppressing MMP secretion and metastasis, established the first defined E3 ligase-mediated regulatory mechanism controlling MYOF protein turnover.\",\n      \"evidence\": \"K48-linkage-specific ubiquitination assays, TRIM8 gain/loss-of-function, MYOF rescue in TRIM8-overexpressing NSCLC cells, xenograft model, MMP secretion assays\",\n      \"pmids\": [\"39934162\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"TRIM8 recognition motif on MYOF not mapped\",\n        \"Whether TRIM8 regulation of MYOF operates in muscle or non-cancer contexts is unknown\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis for MYOF's multi-C2-domain architecture in membrane fusion, the precise domain requirements for its scaffolding of OTUB1/ILF3 and EHD recruitment, and whether its muscle and cancer functions share a common vesicle-trafficking mechanism remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No high-resolution structure of full-length MYOF or individual C2 domains beyond C2A\",\n        \"Functional contributions of C2B–C2F domains are largely uncharacterized\",\n        \"Integration of membrane repair, vesicle trafficking, and scaffolding functions into a unified model is lacking\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [4, 7]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [6, 7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"OTUB1\",\n      \"ILF3\",\n      \"TRIM8\",\n      \"RAB11A\",\n      \"HNRNPLL\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}