{"gene":"VPS25","run_date":"2026-06-11T09:02:06","timeline":{"discoveries":[{"year":2005,"finding":"Drosophila Vps25, as a component of the ESCRT-II complex, is required for endocytic sorting of signaling receptors; loss of vps25 causes endosomal accumulation of the Notch receptor, leading to enhanced Notch signaling, ectopic production of the JAK-STAT ligand Unpaired, and non-autonomous overproliferation of neighboring epithelium.","method":"Drosophila genetic mosaic analysis (loss-of-function clones), immunofluorescence of endosomal Notch accumulation, genetic epistasis with Notch and JAK-STAT pathway components","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal genetic epistasis and direct imaging of endosomal accumulation, independently replicated in two concurrent papers (PMID:16256743 and PMID:16256745)","pmids":["16256743","16256745"],"is_preprint":false},{"year":2005,"finding":"Drosophila Vps25/ESCRT-II loss causes endosomal blockage leading to accumulation of both Notch and Dpp receptors (and associated signaling components), activating both Notch and Dpp receptor signaling, which induces ectopic organizers and overproliferation; when apoptosis is blocked in vps25 mutant cells, tumor-like overgrowths capable of metastasis are formed.","method":"Drosophila genetic mosaic analysis, receptor accumulation assays by immunofluorescence, epistasis with apoptosis inhibitors","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (genetic epistasis, receptor localization, tumor overgrowth phenotype), replicated independently","pmids":["16256745"],"is_preprint":false},{"year":2006,"finding":"In Drosophila vps25 mosaic tissue, failure to downregulate Notch signaling activates the JAK/STAT pathway non-autonomously; vps25 mutant cells undergo apoptosis via Hid and JNK, and Hippo signaling is increased in vps25 clones such that hippo mutants block apoptosis in vps25 clones.","method":"Drosophila genetic screen and mosaic analysis, epistasis with hid, JNK, and hippo pathway mutants, immunofluorescence","journal":"Development (Cambridge, England)","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis across multiple pathways, independent study replicating core Notch/JAK-STAT mechanism and adding Hippo epistasis","pmids":["16611691"],"is_preprint":false},{"year":2004,"finding":"Crystal structure of VPS25 at 3.1 Å resolution reveals that each monomer is composed of two winged-helix domains arranged in tandem, and VPS25 crystallizes in a dimeric form; no conformational changes are detected between unliganded VPS25 and VPS25 within the intact ESCRT-II complex (which contains two VPS25 copies plus one each of Vps22 and Vps36).","method":"X-ray crystallography at 3.1 Å resolution, structural comparison with ESCRT-II complex structure","journal":"BMC structural biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure determination with structural comparison, single lab but definitive structural method","pmids":["15579210"],"is_preprint":false},{"year":2005,"finding":"Human EAP20 (VPS25) directly interacts with CHMP6 (human Vps20/ESCRT-III component); this direct physical interaction is mediated by the N-terminal basic half of CHMP6, as demonstrated by in vitro pull-down with purified recombinant proteins. Co-expression of EAP20 with CHMP6-GFP recruits EAP20 to punctate endosomal structures. CHMP6 overexpression disrupts endosomal cargo sorting (accumulation of ubiquitinated proteins and EGF), suggesting EAP20/VPS25 acts as an acceptor for ESCRT-II on endosomal membranes.","method":"Co-immunoprecipitation of epitope-tagged proteins in HEK-293 cells; in vitro pull-down with purified recombinant proteins from E. coli; fluorescence microscopy of co-expressed proteins in HeLa cells","journal":"The Biochemical journal","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct interaction confirmed by in vitro reconstitution with purified proteins plus co-IP and co-localization, single lab with multiple orthogonal methods","pmids":["15511219"],"is_preprint":false},{"year":2018,"finding":"Xenopus ESCRT-II binds hundreds of mRNAs and the subunit Vps25 directly binds RNA through UV cross-linking; Vps25 specifically recognizes a polypurine (GA-rich) motif. This selective RNA binding was reconstituted in vitro with purified components, establishing Vps25 as the RNA-binding subunit of the ESCRT-II complex.","method":"UV cross-linking, CLIP-Seq (UV cross-linking and immunoprecipitation followed by sequencing), in vitro RNA-binding reconstitution with purified components","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of RNA binding with purified components plus CLIP-Seq motif identification, single lab with multiple orthogonal methods","pmids":["29903915"],"is_preprint":false},{"year":2014,"finding":"Mouse Vps25 (ESCRT-II component) preferentially mediates endosome-dependent degradation/modulation of FGF receptor signaling in limbs; a hypomorphic Vps25 mutation causes aberrant FGFR trafficking and degradation, FGF signaling enhancement, and hyperactivation of the FGF-SHH feedback loop leading to polydactyly, while WNT and BMP signaling remain unperturbed. SHH signaling is not affected in Vps25(ENU/ENU) MEFs, indicating selective receptor trafficking.","method":"ENU-induced mutagenesis, Vps25-null mouse generation, mouse embryonic fibroblast FGFR trafficking assays, signaling pathway analysis","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 / Moderate — hypomorphic and null mouse genetics combined with cellular FGFR trafficking assays and pathway-specific readouts, single lab with multiple orthogonal approaches","pmids":["25373905"],"is_preprint":false},{"year":2011,"finding":"Sprouty 2 (Spry2) binds the ESCRT-II component Eap20 (VPS25); this interaction allows Spry2 to disrupt ESCRT-I interaction with ESCRT-II, thereby facilitating HIV-1 Gag VLP release by preventing the normal sequential handoff from ESCRT-I to ESCRT-II.","method":"Co-immunoprecipitation, VLP release assays in COS-1 cells, co-expression of Spry2 fragments with Eap20, siRNA knockdown of Spry2","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP demonstrating Spry2-Eap20 binding plus functional VLP release assays, single lab, two complementary methods","pmids":["21543492"],"is_preprint":false},{"year":2020,"finding":"EAP20/VPS25 (essential ESCRT-II subunit) is dispensable for HSV-1 replication; siRNA knockdown of EAP20/VPS25 (confirmed by Western blot and quantitative microscopic ESCRT-II functional assays) did not change final HSV-1 yields in single-step replication experiments.","method":"siRNA knockdown, Western blotting, quantitative microscopic ESCRT-II functional assay, single-step viral replication assay","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — siRNA KD confirmed by Western blot and functional ESCRT-II assay with quantitative viral yield readout; finding is a negative result (VPS25 not required for HSV-1)","pmids":["31748394"],"is_preprint":false},{"year":2006,"finding":"Evolutionary analysis of 119 Vps25 orthologs identified two N-terminal PPXY motifs (motif I and II) involved in Vps25p dimerization and interaction with Vps22p and Vps36p within ESCRT-II; Arginine-83 of yeast Vps25p is involved in Vps22p interaction; a highly conserved C-terminal lysine suggests Vps25 may be ubiquitinated.","method":"Comparative sequence analysis of 119 orthologs, cross-referenced with known yeast Vps25p structural/functional data","journal":"BMC evolutionary biology","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational/evolutionary sequence analysis; dimerization and Vps22p interaction residues inferred from yeast data, no direct experimental validation in this study","pmids":["16889659"],"is_preprint":false},{"year":2021,"finding":"VPS25 knockdown in human glioma cells inhibits proliferation, blocks cell cycle at G0/G1, and promotes apoptosis; this is associated with direct modulation of p21, CDK2, and cyclin E expression and inhibition of JAK-STAT activation.","method":"siRNA/shRNA knockdown, cell cycle analysis by flow cytometry, western blotting for cell cycle regulators, transcriptome sequencing","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean KD with defined molecular readouts (cell cycle regulators, JAK-STAT), single lab, single study","pmids":["34863175"],"is_preprint":false},{"year":2026,"finding":"EAP20 (VPS25) participates in transport of internalized PRRSV particles to early endosomes via the clathrin-mediated endocytosis pathway; during viral replication, EAP20 interacts with PRRSV nonstructural proteins Nsp2, Nsp5, and Nsp9, anchors Nsp9 on the perinuclear ER, and cooperates with Nsp2/Nsp5 to form ER-derived double-membrane vesicles.","method":"siRNA knockdown, co-immunoprecipitation, confocal microscopy, viral entry and replication assays","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP plus functional knockdown assays with specific viral replication readouts, single lab, single study","pmids":["41805194"],"is_preprint":false}],"current_model":"VPS25/EAP20 is the winged-helix domain-containing subunit of the ESCRT-II complex (two copies per complex alongside Vps22 and Vps36) that mediates endosomal sorting of ubiquitinated transmembrane receptors into multivesicular bodies; it directly binds the ESCRT-III component CHMP6 to bridge ESCRT-II and ESCRT-III at endosomal membranes, selectively promotes degradation of specific receptors such as FGFRs and Notch to limit their signaling, serves as the RNA-binding subunit of ESCRT-II through recognition of GA-rich polypurine motifs, and its loss causes endosomal receptor accumulation with consequent hyperactivation of Notch, Dpp, and FGF signaling leading to proliferative and developmental defects."},"narrative":{"mechanistic_narrative":"VPS25 (EAP20) is a winged-helix subunit of the ESCRT-II complex that couples endosomal sorting of ubiquitinated transmembrane receptors to the downregulation of developmental signaling pathways [PMID:16256743, PMID:16256745, PMID:15579210]. Structurally, each VPS25 monomer comprises two tandem winged-helix domains and assembles as two copies within ESCRT-II alongside single copies of Vps22 and Vps36, with no conformational change upon complex incorporation [PMID:15579210]. VPS25 directly binds the ESCRT-III component CHMP6 through CHMP6's N-terminal basic region, acting as the acceptor that bridges ESCRT-II to ESCRT-III on endosomal membranes [PMID:15511219]. Loss of VPS25 blocks endosomal sorting and causes accumulation of signaling receptors, leading to hyperactivation of Notch and Dpp signaling, non-autonomous JAK-STAT activation, ectopic organizer formation, and overproliferation that can progress to metastatic tumor-like overgrowths when apoptosis is suppressed [PMID:16256743, PMID:16256745, PMID:16611691]. This receptor regulation is selective: hypomorphic Vps25 specifically perturbs FGFR trafficking and degradation to hyperactivate the FGF-SHH loop while leaving WNT, BMP, and SHH responses intact [PMID:25373905]. Beyond canonical ESCRT function, VPS25 serves as the RNA-binding subunit of ESCRT-II, directly recognizing GA-rich polypurine motifs [PMID:29903915]. VPS25 knockdown in human glioma cells arrests the cell cycle at G0/G1 and promotes apoptosis through modulation of p21, CDK2, and cyclin E and suppression of JAK-STAT signaling [PMID:34863175].","teleology":[{"year":2004,"claim":"Establishing the architecture of VPS25 was needed to understand how it functions within ESCRT-II; the structure revealed a tandem winged-helix fold and defined the two-copy stoichiometry of the complex.","evidence":"X-ray crystallography at 3.1 Å with comparison to the intact ESCRT-II complex","pmids":["15579210"],"confidence":"High","gaps":["Does not address how VPS25 engages cargo or downstream ESCRT-III partners","No structure of the RNA-bound or CHMP6-bound state"]},{"year":2005,"claim":"It was unclear how ESCRT-II connects to the membrane-remodeling ESCRT-III machinery; the discovery that VPS25/EAP20 directly binds CHMP6 defined the molecular bridge between the two complexes.","evidence":"Co-IP in HEK-293, in vitro pull-down with purified recombinant proteins, and co-localization in HeLa cells","pmids":["15511219"],"confidence":"High","gaps":["Does not resolve the structural basis of the VPS25-CHMP6 interface","Functional consequence for cargo sorting inferred from overexpression phenotypes"]},{"year":2005,"claim":"The physiological role of ESCRT-II in tissue signaling was unknown; Drosophila vps25 loss-of-function showed that VPS25 limits Notch and Dpp signaling by sorting receptors into the degradative pathway, linking endosomal trafficking to growth control.","evidence":"Drosophila genetic mosaic analysis, endosomal receptor immunofluorescence, epistasis with Notch/JAK-STAT/apoptosis pathways","pmids":["16256743","16256745"],"confidence":"High","gaps":["Whether receptor accumulation is a direct or indirect consequence of sorting block","Did not identify the specific receptor degradation step VPS25 controls"]},{"year":2006,"claim":"How loss of VPS25 triggers non-autonomous overgrowth versus autonomous death was open; epistasis showed mutant cells activate JAK-STAT non-autonomously while undergoing Hid/JNK apoptosis under Hippo control.","evidence":"Drosophila genetic screen, mosaic analysis, and epistasis with hid, JNK, and hippo mutants","pmids":["16611691"],"confidence":"High","gaps":["Molecular link between receptor accumulation and Hippo pathway activation not defined","Conservation of the apoptotic/proliferative balance in mammals not tested"]},{"year":2006,"claim":"Sequence determinants of VPS25 assembly were uncharacterized; comparative analysis proposed N-terminal PPXY motifs and Arg-83 in dimerization and Vps22/Vps36 contacts and a conserved C-terminal lysine as a possible ubiquitination site.","evidence":"Comparative sequence analysis of 119 orthologs cross-referenced to yeast structural data","pmids":["16889659"],"confidence":"Low","gaps":["Computational inference without direct experimental validation in this study","Predicted ubiquitination of the C-terminal lysine untested","PPXY motif function not confirmed by mutagenesis here"]},{"year":2011,"claim":"Whether VPS25 could be co-opted to redirect ESCRT handoff was unknown; Spry2 was shown to bind EAP20 and disrupt the ESCRT-I-to-ESCRT-II handoff, modulating HIV-1 Gag VLP release.","evidence":"Co-IP, VLP release assays in COS-1 cells, Spry2 fragment co-expression, and Spry2 siRNA knockdown","pmids":["21543492"],"confidence":"Medium","gaps":["Single lab with two complementary methods, no reciprocal structural validation","Physiological relevance of Spry2-EAP20 binding outside viral budding unclear"]},{"year":2014,"claim":"Whether ESCRT-II receptor regulation is general or selective was unresolved; mouse Vps25 hypomorphs showed it preferentially controls FGFR trafficking and the FGF-SHH loop without affecting WNT, BMP, or SHH responses.","evidence":"ENU mutagenesis, Vps25-null mice, MEF FGFR trafficking assays, and pathway-specific signaling readouts","pmids":["25373905"],"confidence":"High","gaps":["Mechanistic basis of receptor selectivity not defined","Whether selectivity reflects cargo recognition or differential degradation kinetics unknown"]},{"year":2018,"claim":"A non-trafficking activity of ESCRT-II was undefined; reconstitution and CLIP-Seq established VPS25 as the RNA-binding subunit recognizing GA-rich polypurine motifs.","evidence":"UV cross-linking, CLIP-Seq motif identification, and in vitro RNA-binding reconstitution with purified components","pmids":["29903915"],"confidence":"High","gaps":["Functional consequence of VPS25-RNA binding for the bound mRNAs not established","Relationship between RNA binding and endosomal sorting unknown"]},{"year":2020,"claim":"Whether ESCRT-II is required for all enveloped-virus egress was tested; VPS25 knockdown did not alter HSV-1 yields, showing it is dispensable for HSV-1 replication.","evidence":"siRNA knockdown confirmed by Western blot and quantitative ESCRT-II functional assay, single-step viral replication assay","pmids":["31748394"],"confidence":"Medium","gaps":["Negative result does not exclude redundancy with other ESCRT routes","Does not address other herpesviruses or cargo classes"]},{"year":2021,"claim":"The relevance of VPS25 to human cancer cell proliferation was untested; knockdown in glioma cells arrested cells at G0/G1 and induced apoptosis via p21/CDK2/cyclin E and JAK-STAT modulation.","evidence":"siRNA/shRNA knockdown, flow cytometry cell cycle analysis, western blotting, and transcriptome sequencing","pmids":["34863175"],"confidence":"Medium","gaps":["Whether cell cycle effects are downstream of endosomal sorting or RNA binding not resolved","Single cancer cell type, single study"]},{"year":2026,"claim":"Whether VPS25 supports specific viral life cycles was further probed; EAP20 was shown to aid PRRSV endosomal entry and to interact with Nsp2/Nsp5/Nsp9 to build ER-derived double-membrane replication vesicles.","evidence":"siRNA knockdown, co-IP, confocal microscopy, and viral entry/replication assays","pmids":["41805194"],"confidence":"Medium","gaps":["Direct vs indirect nature of Nsp interactions not structurally validated","Single lab, single study"]},{"year":null,"claim":"How VPS25 achieves receptor selectivity and how its RNA-binding activity integrates with its canonical ESCRT sorting role remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of cargo selectivity","Functional output of GA-rich mRNA binding undefined","Link between ESCRT sorting and cell cycle control in human cells unestablished"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[5]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[4]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[4,0]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[11]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,6]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[1,6]}],"complexes":["ESCRT-II"],"partners":["CHMP6","VPS22","VPS36","SPRY2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BRG1","full_name":"Vacuolar protein-sorting-associated protein 25","aliases":["Dermal papilla-derived protein 9","ELL-associated protein of 20 kDa","ESCRT-II complex subunit VPS25"],"length_aa":176,"mass_kda":20.7,"function":"Component of the ESCRT-II complex (endosomal sorting complex required for transport II), which is required for multivesicular body (MVB) formation and sorting of endosomal cargo proteins into MVBs. The MVB pathway mediates delivery of transmembrane proteins into the lumen of the lysosome for degradation. The ESCRT-II complex is probably involved in the recruitment of the ESCRT-III complex. The ESCRT-II complex may also play a role in transcription regulation, possibly via its interaction with ELL. The ESCRT-II complex may be involved in facilitating the budding of certain RNA viruses","subcellular_location":"Cytoplasm; Endosome membrane; Nucleus, nucleoplasm","url":"https://www.uniprot.org/uniprotkb/Q9BRG1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/VPS25","classification":"Common Essential","n_dependent_lines":1206,"n_total_lines":1208,"dependency_fraction":0.9983443708609272},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000131475","cell_line_id":"CID000788","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"nucleoplasm","grade":3}],"interactors":[{"gene":"SNF8","stoichiometry":10.0},{"gene":"VPS36","stoichiometry":10.0},{"gene":"PMVK","stoichiometry":0.2},{"gene":"VPS37B","stoichiometry":0.2},{"gene":"TSG101","stoichiometry":0.2},{"gene":"VPS28","stoichiometry":0.2},{"gene":"MVB12A","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000788","total_profiled":1310},"omim":[{"mim_id":"620784","title":"NEURODEVELOPMENTAL DISORDER PLUS OPTIC ATROPHY; NEDOA","url":"https://www.omim.org/entry/620784"},{"mim_id":"620783","title":"DEVELOPMENTAL AND EPILEPTIC ENCEPHALOPATHY 115; DEE115","url":"https://www.omim.org/entry/620783"},{"mim_id":"611130","title":"CHARGED MULTIVESICULAR BODY PROTEIN 7; CHMP7","url":"https://www.omim.org/entry/611130"},{"mim_id":"610907","title":"VACUOLAR PROTEIN SORTING 25 HOMOLOG; VPS25","url":"https://www.omim.org/entry/610907"},{"mim_id":"610904","title":"SNF8 SUBUNIT OF ESCRIT-II; SNF8","url":"https://www.omim.org/entry/610904"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Vesicles","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/VPS25"},"hgnc":{"alias_symbol":["MGC10540","EAP20","DERP9"],"prev_symbol":[]},"alphafold":{"accession":"Q9BRG1","domains":[{"cath_id":"1.10.10.570","chopping":"9-98","consensus_level":"high","plddt":94.9079,"start":9,"end":98},{"cath_id":"1.10.10.10","chopping":"103-174","consensus_level":"high","plddt":92.4976,"start":103,"end":174}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRG1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRG1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRG1-F1-predicted_aligned_error_v6.png","plddt_mean":92.94},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=VPS25","jax_strain_url":"https://www.jax.org/strain/search?query=VPS25"},"sequence":{"accession":"Q9BRG1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BRG1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BRG1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRG1"}},"corpus_meta":[{"pmid":"16256743","id":"PMC_16256743","title":"The Drosophila tumor suppressor vps25 prevents nonautonomous overproliferation by regulating notch trafficking.","date":"2005","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/16256743","citation_count":315,"is_preprint":false},{"pmid":"16256745","id":"PMC_16256745","title":"Tumor suppressor properties of the ESCRT-II complex component Vps25 in Drosophila.","date":"2005","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/16256745","citation_count":285,"is_preprint":false},{"pmid":"16611691","id":"PMC_16611691","title":"vps25 mosaics display non-autonomous cell survival and overgrowth, and autonomous apoptosis.","date":"2006","source":"Development (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/16611691","citation_count":141,"is_preprint":false},{"pmid":"15511219","id":"PMC_15511219","title":"Human CHMP6, a myristoylated ESCRT-III protein, interacts directly with an ESCRT-II component EAP20 and regulates endosomal cargo sorting.","date":"2005","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/15511219","citation_count":103,"is_preprint":false},{"pmid":"34863175","id":"PMC_34863175","title":"YTHDC1-mediated VPS25 regulates cell cycle by targeting JAK-STAT signaling in human glioma cells.","date":"2021","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/34863175","citation_count":32,"is_preprint":false},{"pmid":"31748394","id":"PMC_31748394","title":"The ESCRT-II Subunit EAP20/VPS25 and the Bro1 Domain Proteins HD-PTP and BROX Are Individually Dispensable for Herpes Simplex Virus 1 Replication.","date":"2020","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/31748394","citation_count":17,"is_preprint":false},{"pmid":"25373905","id":"PMC_25373905","title":"ESCRT-II/Vps25 constrains digit number by endosome-mediated selective modulation of FGF-SHH signaling.","date":"2014","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/25373905","citation_count":15,"is_preprint":false},{"pmid":"21543492","id":"PMC_21543492","title":"Sprouty 2 binds ESCRT-II factor Eap20 and facilitates HIV-1 gag release.","date":"2011","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/21543492","citation_count":13,"is_preprint":false},{"pmid":"15579210","id":"PMC_15579210","title":"Crystal structure of subunit VPS25 of the endosomal trafficking complex ESCRT-II.","date":"2004","source":"BMC structural biology","url":"https://pubmed.ncbi.nlm.nih.gov/15579210","citation_count":11,"is_preprint":false},{"pmid":"16889659","id":"PMC_16889659","title":"Genetic structure and evolution of the Vps25 family, a yeast ESCRT-II component.","date":"2006","source":"BMC evolutionary biology","url":"https://pubmed.ncbi.nlm.nih.gov/16889659","citation_count":8,"is_preprint":false},{"pmid":"29903915","id":"PMC_29903915","title":"The RNA-binding complex ESCRT-II in Xenopus laevis eggs recognizes purine-rich sequences through its subunit, Vps25.","date":"2018","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/29903915","citation_count":5,"is_preprint":false},{"pmid":"40149859","id":"PMC_40149859","title":"VPS25 Promotes an Immunosuppressive Microenvironment in Head and Neck Squamous Cell Carcinoma.","date":"2025","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40149859","citation_count":1,"is_preprint":false},{"pmid":"37766995","id":"PMC_37766995","title":"Crosstalk between integrin/FAK and Crk/Vps25 governs invasion of bovine mammary epithelial cells by S. agalactiae.","date":"2023","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/37766995","citation_count":1,"is_preprint":false},{"pmid":"38670586","id":"PMC_38670586","title":"Germline MYOF1::WNK4 and VPS25::MYOF1 Chimeras Generated by the Constitutional Translocation t(17;19)(q21;p13) in Two Siblings With Myelodysplastic Syndrome.","date":"2024","source":"Cancer genomics & proteomics","url":"https://pubmed.ncbi.nlm.nih.gov/38670586","citation_count":1,"is_preprint":false},{"pmid":"41805194","id":"PMC_41805194","title":"Porcine reproductive and respiratory syndrome virus exploits ESCRT-II subunit EAP20 for entry and replication.","date":"2026","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/41805194","citation_count":0,"is_preprint":false},{"pmid":"41448467","id":"PMC_41448467","title":"VPS25 alleviates depression-like behavior in rats by inhibiting apoptosis in the hippocampus.","date":"2025","source":"Brain research bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/41448467","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.11.26.690820","title":"Genome-wide association mapping for grain and forage quality traits in a subtropical oat germplasm collection adapted to highland regions of Eastern Africa","date":"2025-11-27","source":"bioRxiv","url":"https://doi.org/10.1101/2025.11.26.690820","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10823,"output_tokens":3310,"usd":0.041059,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10741,"output_tokens":3848,"usd":0.074953,"stage2_stop_reason":"end_turn"},"total_usd":0.116012,"stage1_batch_id":"msgbatch_01UT1AahFPCAyn2uVweS7A9z","stage2_batch_id":"msgbatch_01HuKWtKNT4Pqj1bc8eEP5RV","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"Drosophila Vps25, as a component of the ESCRT-II complex, is required for endocytic sorting of signaling receptors; loss of vps25 causes endosomal accumulation of the Notch receptor, leading to enhanced Notch signaling, ectopic production of the JAK-STAT ligand Unpaired, and non-autonomous overproliferation of neighboring epithelium.\",\n      \"method\": \"Drosophila genetic mosaic analysis (loss-of-function clones), immunofluorescence of endosomal Notch accumulation, genetic epistasis with Notch and JAK-STAT pathway components\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal genetic epistasis and direct imaging of endosomal accumulation, independently replicated in two concurrent papers (PMID:16256743 and PMID:16256745)\",\n      \"pmids\": [\"16256743\", \"16256745\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Drosophila Vps25/ESCRT-II loss causes endosomal blockage leading to accumulation of both Notch and Dpp receptors (and associated signaling components), activating both Notch and Dpp receptor signaling, which induces ectopic organizers and overproliferation; when apoptosis is blocked in vps25 mutant cells, tumor-like overgrowths capable of metastasis are formed.\",\n      \"method\": \"Drosophila genetic mosaic analysis, receptor accumulation assays by immunofluorescence, epistasis with apoptosis inhibitors\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (genetic epistasis, receptor localization, tumor overgrowth phenotype), replicated independently\",\n      \"pmids\": [\"16256745\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"In Drosophila vps25 mosaic tissue, failure to downregulate Notch signaling activates the JAK/STAT pathway non-autonomously; vps25 mutant cells undergo apoptosis via Hid and JNK, and Hippo signaling is increased in vps25 clones such that hippo mutants block apoptosis in vps25 clones.\",\n      \"method\": \"Drosophila genetic screen and mosaic analysis, epistasis with hid, JNK, and hippo pathway mutants, immunofluorescence\",\n      \"journal\": \"Development (Cambridge, England)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis across multiple pathways, independent study replicating core Notch/JAK-STAT mechanism and adding Hippo epistasis\",\n      \"pmids\": [\"16611691\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Crystal structure of VPS25 at 3.1 Å resolution reveals that each monomer is composed of two winged-helix domains arranged in tandem, and VPS25 crystallizes in a dimeric form; no conformational changes are detected between unliganded VPS25 and VPS25 within the intact ESCRT-II complex (which contains two VPS25 copies plus one each of Vps22 and Vps36).\",\n      \"method\": \"X-ray crystallography at 3.1 Å resolution, structural comparison with ESCRT-II complex structure\",\n      \"journal\": \"BMC structural biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure determination with structural comparison, single lab but definitive structural method\",\n      \"pmids\": [\"15579210\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Human EAP20 (VPS25) directly interacts with CHMP6 (human Vps20/ESCRT-III component); this direct physical interaction is mediated by the N-terminal basic half of CHMP6, as demonstrated by in vitro pull-down with purified recombinant proteins. Co-expression of EAP20 with CHMP6-GFP recruits EAP20 to punctate endosomal structures. CHMP6 overexpression disrupts endosomal cargo sorting (accumulation of ubiquitinated proteins and EGF), suggesting EAP20/VPS25 acts as an acceptor for ESCRT-II on endosomal membranes.\",\n      \"method\": \"Co-immunoprecipitation of epitope-tagged proteins in HEK-293 cells; in vitro pull-down with purified recombinant proteins from E. coli; fluorescence microscopy of co-expressed proteins in HeLa cells\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct interaction confirmed by in vitro reconstitution with purified proteins plus co-IP and co-localization, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"15511219\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Xenopus ESCRT-II binds hundreds of mRNAs and the subunit Vps25 directly binds RNA through UV cross-linking; Vps25 specifically recognizes a polypurine (GA-rich) motif. This selective RNA binding was reconstituted in vitro with purified components, establishing Vps25 as the RNA-binding subunit of the ESCRT-II complex.\",\n      \"method\": \"UV cross-linking, CLIP-Seq (UV cross-linking and immunoprecipitation followed by sequencing), in vitro RNA-binding reconstitution with purified components\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of RNA binding with purified components plus CLIP-Seq motif identification, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"29903915\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Mouse Vps25 (ESCRT-II component) preferentially mediates endosome-dependent degradation/modulation of FGF receptor signaling in limbs; a hypomorphic Vps25 mutation causes aberrant FGFR trafficking and degradation, FGF signaling enhancement, and hyperactivation of the FGF-SHH feedback loop leading to polydactyly, while WNT and BMP signaling remain unperturbed. SHH signaling is not affected in Vps25(ENU/ENU) MEFs, indicating selective receptor trafficking.\",\n      \"method\": \"ENU-induced mutagenesis, Vps25-null mouse generation, mouse embryonic fibroblast FGFR trafficking assays, signaling pathway analysis\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — hypomorphic and null mouse genetics combined with cellular FGFR trafficking assays and pathway-specific readouts, single lab with multiple orthogonal approaches\",\n      \"pmids\": [\"25373905\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Sprouty 2 (Spry2) binds the ESCRT-II component Eap20 (VPS25); this interaction allows Spry2 to disrupt ESCRT-I interaction with ESCRT-II, thereby facilitating HIV-1 Gag VLP release by preventing the normal sequential handoff from ESCRT-I to ESCRT-II.\",\n      \"method\": \"Co-immunoprecipitation, VLP release assays in COS-1 cells, co-expression of Spry2 fragments with Eap20, siRNA knockdown of Spry2\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP demonstrating Spry2-Eap20 binding plus functional VLP release assays, single lab, two complementary methods\",\n      \"pmids\": [\"21543492\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EAP20/VPS25 (essential ESCRT-II subunit) is dispensable for HSV-1 replication; siRNA knockdown of EAP20/VPS25 (confirmed by Western blot and quantitative microscopic ESCRT-II functional assays) did not change final HSV-1 yields in single-step replication experiments.\",\n      \"method\": \"siRNA knockdown, Western blotting, quantitative microscopic ESCRT-II functional assay, single-step viral replication assay\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — siRNA KD confirmed by Western blot and functional ESCRT-II assay with quantitative viral yield readout; finding is a negative result (VPS25 not required for HSV-1)\",\n      \"pmids\": [\"31748394\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Evolutionary analysis of 119 Vps25 orthologs identified two N-terminal PPXY motifs (motif I and II) involved in Vps25p dimerization and interaction with Vps22p and Vps36p within ESCRT-II; Arginine-83 of yeast Vps25p is involved in Vps22p interaction; a highly conserved C-terminal lysine suggests Vps25 may be ubiquitinated.\",\n      \"method\": \"Comparative sequence analysis of 119 orthologs, cross-referenced with known yeast Vps25p structural/functional data\",\n      \"journal\": \"BMC evolutionary biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational/evolutionary sequence analysis; dimerization and Vps22p interaction residues inferred from yeast data, no direct experimental validation in this study\",\n      \"pmids\": [\"16889659\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"VPS25 knockdown in human glioma cells inhibits proliferation, blocks cell cycle at G0/G1, and promotes apoptosis; this is associated with direct modulation of p21, CDK2, and cyclin E expression and inhibition of JAK-STAT activation.\",\n      \"method\": \"siRNA/shRNA knockdown, cell cycle analysis by flow cytometry, western blotting for cell cycle regulators, transcriptome sequencing\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean KD with defined molecular readouts (cell cycle regulators, JAK-STAT), single lab, single study\",\n      \"pmids\": [\"34863175\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"EAP20 (VPS25) participates in transport of internalized PRRSV particles to early endosomes via the clathrin-mediated endocytosis pathway; during viral replication, EAP20 interacts with PRRSV nonstructural proteins Nsp2, Nsp5, and Nsp9, anchors Nsp9 on the perinuclear ER, and cooperates with Nsp2/Nsp5 to form ER-derived double-membrane vesicles.\",\n      \"method\": \"siRNA knockdown, co-immunoprecipitation, confocal microscopy, viral entry and replication assays\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP plus functional knockdown assays with specific viral replication readouts, single lab, single study\",\n      \"pmids\": [\"41805194\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"VPS25/EAP20 is the winged-helix domain-containing subunit of the ESCRT-II complex (two copies per complex alongside Vps22 and Vps36) that mediates endosomal sorting of ubiquitinated transmembrane receptors into multivesicular bodies; it directly binds the ESCRT-III component CHMP6 to bridge ESCRT-II and ESCRT-III at endosomal membranes, selectively promotes degradation of specific receptors such as FGFRs and Notch to limit their signaling, serves as the RNA-binding subunit of ESCRT-II through recognition of GA-rich polypurine motifs, and its loss causes endosomal receptor accumulation with consequent hyperactivation of Notch, Dpp, and FGF signaling leading to proliferative and developmental defects.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"VPS25 (EAP20) is a winged-helix subunit of the ESCRT-II complex that couples endosomal sorting of ubiquitinated transmembrane receptors to the downregulation of developmental signaling pathways [#0, #3]. Structurally, each VPS25 monomer comprises two tandem winged-helix domains and assembles as two copies within ESCRT-II alongside single copies of Vps22 and Vps36, with no conformational change upon complex incorporation [#3]. VPS25 directly binds the ESCRT-III component CHMP6 through CHMP6's N-terminal basic region, acting as the acceptor that bridges ESCRT-II to ESCRT-III on endosomal membranes [#4]. Loss of VPS25 blocks endosomal sorting and causes accumulation of signaling receptors, leading to hyperactivation of Notch and Dpp signaling, non-autonomous JAK-STAT activation, ectopic organizer formation, and overproliferation that can progress to metastatic tumor-like overgrowths when apoptosis is suppressed [#0, #1, #2]. This receptor regulation is selective: hypomorphic Vps25 specifically perturbs FGFR trafficking and degradation to hyperactivate the FGF-SHH loop while leaving WNT, BMP, and SHH responses intact [#6]. Beyond canonical ESCRT function, VPS25 serves as the RNA-binding subunit of ESCRT-II, directly recognizing GA-rich polypurine motifs [#5]. VPS25 knockdown in human glioma cells arrests the cell cycle at G0/G1 and promotes apoptosis through modulation of p21, CDK2, and cyclin E and suppression of JAK-STAT signaling [#10].\",\n  \"teleology\": [\n    {\n      \"year\": 2004,\n      \"claim\": \"Establishing the architecture of VPS25 was needed to understand how it functions within ESCRT-II; the structure revealed a tandem winged-helix fold and defined the two-copy stoichiometry of the complex.\",\n      \"evidence\": \"X-ray crystallography at 3.1 Å with comparison to the intact ESCRT-II complex\",\n      \"pmids\": [\"15579210\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not address how VPS25 engages cargo or downstream ESCRT-III partners\", \"No structure of the RNA-bound or CHMP6-bound state\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"It was unclear how ESCRT-II connects to the membrane-remodeling ESCRT-III machinery; the discovery that VPS25/EAP20 directly binds CHMP6 defined the molecular bridge between the two complexes.\",\n      \"evidence\": \"Co-IP in HEK-293, in vitro pull-down with purified recombinant proteins, and co-localization in HeLa cells\",\n      \"pmids\": [\"15511219\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not resolve the structural basis of the VPS25-CHMP6 interface\", \"Functional consequence for cargo sorting inferred from overexpression phenotypes\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"The physiological role of ESCRT-II in tissue signaling was unknown; Drosophila vps25 loss-of-function showed that VPS25 limits Notch and Dpp signaling by sorting receptors into the degradative pathway, linking endosomal trafficking to growth control.\",\n      \"evidence\": \"Drosophila genetic mosaic analysis, endosomal receptor immunofluorescence, epistasis with Notch/JAK-STAT/apoptosis pathways\",\n      \"pmids\": [\"16256743\", \"16256745\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether receptor accumulation is a direct or indirect consequence of sorting block\", \"Did not identify the specific receptor degradation step VPS25 controls\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"How loss of VPS25 triggers non-autonomous overgrowth versus autonomous death was open; epistasis showed mutant cells activate JAK-STAT non-autonomously while undergoing Hid/JNK apoptosis under Hippo control.\",\n      \"evidence\": \"Drosophila genetic screen, mosaic analysis, and epistasis with hid, JNK, and hippo mutants\",\n      \"pmids\": [\"16611691\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular link between receptor accumulation and Hippo pathway activation not defined\", \"Conservation of the apoptotic/proliferative balance in mammals not tested\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Sequence determinants of VPS25 assembly were uncharacterized; comparative analysis proposed N-terminal PPXY motifs and Arg-83 in dimerization and Vps22/Vps36 contacts and a conserved C-terminal lysine as a possible ubiquitination site.\",\n      \"evidence\": \"Comparative sequence analysis of 119 orthologs cross-referenced to yeast structural data\",\n      \"pmids\": [\"16889659\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Computational inference without direct experimental validation in this study\", \"Predicted ubiquitination of the C-terminal lysine untested\", \"PPXY motif function not confirmed by mutagenesis here\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Whether VPS25 could be co-opted to redirect ESCRT handoff was unknown; Spry2 was shown to bind EAP20 and disrupt the ESCRT-I-to-ESCRT-II handoff, modulating HIV-1 Gag VLP release.\",\n      \"evidence\": \"Co-IP, VLP release assays in COS-1 cells, Spry2 fragment co-expression, and Spry2 siRNA knockdown\",\n      \"pmids\": [\"21543492\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab with two complementary methods, no reciprocal structural validation\", \"Physiological relevance of Spry2-EAP20 binding outside viral budding unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Whether ESCRT-II receptor regulation is general or selective was unresolved; mouse Vps25 hypomorphs showed it preferentially controls FGFR trafficking and the FGF-SHH loop without affecting WNT, BMP, or SHH responses.\",\n      \"evidence\": \"ENU mutagenesis, Vps25-null mice, MEF FGFR trafficking assays, and pathway-specific signaling readouts\",\n      \"pmids\": [\"25373905\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanistic basis of receptor selectivity not defined\", \"Whether selectivity reflects cargo recognition or differential degradation kinetics unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"A non-trafficking activity of ESCRT-II was undefined; reconstitution and CLIP-Seq established VPS25 as the RNA-binding subunit recognizing GA-rich polypurine motifs.\",\n      \"evidence\": \"UV cross-linking, CLIP-Seq motif identification, and in vitro RNA-binding reconstitution with purified components\",\n      \"pmids\": [\"29903915\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of VPS25-RNA binding for the bound mRNAs not established\", \"Relationship between RNA binding and endosomal sorting unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Whether ESCRT-II is required for all enveloped-virus egress was tested; VPS25 knockdown did not alter HSV-1 yields, showing it is dispensable for HSV-1 replication.\",\n      \"evidence\": \"siRNA knockdown confirmed by Western blot and quantitative ESCRT-II functional assay, single-step viral replication assay\",\n      \"pmids\": [\"31748394\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Negative result does not exclude redundancy with other ESCRT routes\", \"Does not address other herpesviruses or cargo classes\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"The relevance of VPS25 to human cancer cell proliferation was untested; knockdown in glioma cells arrested cells at G0/G1 and induced apoptosis via p21/CDK2/cyclin E and JAK-STAT modulation.\",\n      \"evidence\": \"siRNA/shRNA knockdown, flow cytometry cell cycle analysis, western blotting, and transcriptome sequencing\",\n      \"pmids\": [\"34863175\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether cell cycle effects are downstream of endosomal sorting or RNA binding not resolved\", \"Single cancer cell type, single study\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Whether VPS25 supports specific viral life cycles was further probed; EAP20 was shown to aid PRRSV endosomal entry and to interact with Nsp2/Nsp5/Nsp9 to build ER-derived double-membrane replication vesicles.\",\n      \"evidence\": \"siRNA knockdown, co-IP, confocal microscopy, and viral entry/replication assays\",\n      \"pmids\": [\"41805194\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct vs indirect nature of Nsp interactions not structurally validated\", \"Single lab, single study\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How VPS25 achieves receptor selectivity and how its RNA-binding activity integrates with its canonical ESCRT sorting role remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of cargo selectivity\", \"Functional output of GA-rich mRNA binding undefined\", \"Link between ESCRT sorting and cell cycle control in human cells unestablished\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [4, 0]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 6]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [1, 6]}\n    ],\n    \"complexes\": [\"ESCRT-II\"],\n    \"partners\": [\"CHMP6\", \"Vps22\", \"Vps36\", \"Spry2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":7,"faith_pct":85.71428571428571}}