{"gene":"RMND5A","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2007,"finding":"RMND5A (p44CTLH) was identified as a component of the mammalian CTLH complex, along with RanBPM, Muskelin, p48EMLP, ARMC8α, and ARMC8β. Each component contains LisH/CTLH motifs. Co-immunoprecipitation and pull-down assays with bacterially-expressed Twa1 confirmed in vivo and in vitro associations of all complex members.","method":"Tandem MS immunoprecipitation, co-immunoprecipitation (Co-IP), and in vitro pull-down assay with bacterially-expressed Twa1","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple orthogonal methods (MS, Co-IP, pull-down) in a single lab; complex membership established but no functional readout specifically for RMND5A","pmids":["17467196"],"is_preprint":false},{"year":2008,"finding":"The yeast ortholog Gid2/Rmd5 contains a degenerate RING finger domain and provides E3 ubiquitin ligase activity to the Gid complex; mutation of the degenerate RING domain abolishes fructose-1,6-bisphosphatase (FBPase) polyubiquitination and degradation in vivo, and heterologous GST-Gid2 expression leads to polyubiquitination in vitro.","method":"In vitro ubiquitination assay (GST-Gid2 expression), RING domain mutagenesis, in vivo degradation assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro ubiquitination assay plus active-site mutagenesis plus in vivo degradation phenotype in yeast ortholog; multiple orthogonal methods in one study","pmids":["18508925"],"is_preprint":false},{"year":2011,"finding":"Yeast Gid2/Rmd5 physically interacts with a second RING finger subunit Gid9/Fyv10 within the Gid complex; mutation of Gid9's RING finger abolishes polyubiquitylation and degradation of three gluconeogenic enzymes, indicating that both RING subunits are required for full E3 ligase activity.","method":"Co-immunoprecipitation, RING domain mutagenesis, in vivo ubiquitination/degradation assay","journal":"FEBS letters","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — reciprocal interaction confirmed, active-site mutagenesis with functional readout in yeast ortholog, single lab with multiple orthogonal methods","pmids":["22044534"],"is_preprint":false},{"year":2012,"finding":"Domain-deletion and point-mutation analysis of yeast Gid complex subunits (including Gid2/Rmd5) revealed the topology of subunit interactions; LisH, CTLH, and SPRY domains in individual Gid proteins mediate specific subunit–subunit contacts within the E3 ubiquitin ligase complex.","method":"Domain deletion/mutagenesis followed by co-immunoprecipitation to map subunit interactions","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic domain mutagenesis with Co-IP readout in yeast ortholog, single lab","pmids":["22645139"],"is_preprint":false},{"year":2013,"finding":"RMND5A was identified as a direct target of miR-138 in HeLa cells; overexpression of miR-138 downregulated RMND5A protein, which in turn reduced Exportin-5 stability and decreased pre-miRNA nuclear export, and also significantly inhibited HeLa cell migration.","method":"miR-138 overexpression, Western blot (RMND5A and Exportin-5 levels), pre-miRNA export assay, wound-healing migration assay","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — multiple functional readouts (protein stability, RNA export, migration) in a single lab; direct experimental validation of miR-138→RMND5A→Exportin-5 pathway","pmids":["24057215"],"is_preprint":false},{"year":2017,"finding":"Downregulation of RMND5A (along with muskelin) decreased acetylation of the HDAC6 substrate α-tubulin, demonstrating that the CTLH complex (via RMND5A) contributes to inhibition of HDAC6 deacetylase activity and consequently restricts cell migration.","method":"Stable knockdown cell lines, Western blot for acetylated α-tubulin, wound-healing migration assay, co-immunoprecipitation","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — functional knockdown with two orthogonal readouts (biochemical activity and cell migration), single lab","pmids":["28668087"],"is_preprint":false},{"year":2019,"finding":"The human CTLH complex immunoprecipitated from cells comprises RanBPM, ARMC8α/β, muskelin, WDR26, GID4, RMND5A, and MAEA. RMND5A and MAEA protein levels are interdependent. In vitro ubiquitination assays showed E3 ligase activity dependent on both RMND5A and MAEA. Recombinant RMND5A mediates K48 and K63 poly-ubiquitin chains and pairs with UBE2D1, UBE2D2, and UBE2D3 E2 enzymes. Muskelin ubiquitination is RMND5A-dependent, and muskelin protein levels increase in RMND5A KO cells in a proteasome-dependent manner.","method":"Co-immunoprecipitation, in vitro ubiquitination assay with recombinant RMND5A, RMND5A knockout cells, ubiquitin linkage-specific analysis, proteasome inhibitor treatment","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution with recombinant protein, active-site-dependent assay, KO cellular validation, multiple orthogonal methods in one rigorous study","pmids":["31285494"],"is_preprint":false},{"year":2019,"finding":"The CTLH complex promotes c-Raf ubiquitination and proteasome-dependent degradation; depletion of RMND5A (or RanBPM) results in enhanced cell proliferation and tumor growth. A-Raf and B-Raf levels are also regulated by the complex, indicating a common Raf family regulation.","method":"RMND5A depletion, Western blot for c-Raf/A-Raf/B-Raf levels, ubiquitination assay, proliferation assay, mouse tumor model","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO/KD with multiple functional and biochemical readouts, in vivo mouse model, single lab","pmids":["30795516"],"is_preprint":false},{"year":2021,"finding":"Proteomic and diGLY-enriched ubiquitinome analysis in CTLH-depleted HeLa cells identified glycolysis enzymes PKM2 and LDHA as RMND5A-dependent ubiquitination substrates. Reduced polyubiquitination of PKM2 and LDHA was validated in RMND5A-depleted cells; their enzymatic activities were increased without changes in protein levels, and RanBPM-deficient cells exhibited enhanced glycolysis, uncovering a non-degradative ubiquitination role for RMND5A/CTLH.","method":"Mass spectrometry-based global proteomics, diGLY-enriched ubiquitinome profiling, RanBPM/RMND5A depletion, enzymatic activity assays (PKM2, LDHA), glycolysis metabolic assay","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal mass spectrometry approaches plus direct biochemical validation of substrate ubiquitination and enzyme activity, single lab with comprehensive methodology","pmids":["34383978"],"is_preprint":false},{"year":2022,"finding":"RMND5A and MAEA (GID complex RING subunits) mediate ubiquitin-proteasome-dependent degradation of the oncopeptide MBOP encoded by LINC01234 in colorectal cancer cells, as shown by immunoprecipitation experiments.","method":"Co-immunoprecipitation, Western blot, proteasome inhibitor treatment","journal":"Cancers","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP experiment identifying RMND5A as one of two E3 ligases for a specific substrate, single lab, limited mechanistic follow-up","pmids":["35565466"],"is_preprint":false},{"year":2021,"finding":"RMND5A overexpression significantly increased cell migration in pancreatic adenocarcinoma cell lines (AsPC-1 and PANC-1), and miR-590-5p-mediated downregulation of RMND5A decreased this migratory ability, placing RMND5A upstream of migration-promoting signaling in PAAD cells.","method":"Wound-healing migration assay, RMND5A overexpression, miR-590-5p overexpression, Western blot","journal":"Oncology letters","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, phenotypic rescue experiment but no direct molecular mechanism identified for how RMND5A promotes migration","pmids":["34079591"],"is_preprint":false},{"year":2024,"finding":"Overexpression of RMND5A in HUVECs reduced proliferation, migration, and tube formation by inhibiting ERK and NF-κB pathway activation. OSCC-derived exosomal miR-21 suppresses RMND5A expression in endothelial cells to promote angiogenesis.","method":"Lentiviral RMND5A overexpression, MTT assay, wound-healing assay, tube formation assay, Western blot (ERK/NF-κB), exosome co-culture","journal":"BMC oral health","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, overexpression-based phenotypic assay without direct substrate identification or molecular mechanism for ERK/NF-κB regulation by RMND5A","pmids":["38229133"],"is_preprint":false},{"year":2025,"finding":"RMND5A knockdown in human neural stem/precursor cells (hNS/PCs) decreased proliferation and promoted neuronal differentiation, associated with activation of Wnt signaling and suppression of mTOR signaling, identifying RMND5A as required for hNS/PC self-renewal.","method":"CRISPR/Cas9 knockout screen, shRNA knockdown, proliferation and differentiation assays, Western blot/pathway analysis (Wnt, mTOR)","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — CRISPR screen plus targeted KD with multiple functional readouts (proliferation, differentiation, pathway activation), single lab","pmids":["40377017"],"is_preprint":false},{"year":2025,"finding":"The GID complex (via MAEA and RMND5A subunits) ubiquitinates and degrades β-Catenin independently of βTrCP when GSK3β is suppressed. Wnt stimulation promotes GSK3β–GID complex interaction, disrupting the MAEA–β-Catenin association and thereby stabilizing β-Catenin. Knockdown of MAEA and RMND5A rescued β-Catenin levels in GSK3β-knockdown cells.","method":"GSK3β knockdown, MAEA/RMND5A knockdown, Western blot for β-Catenin ubiquitination and protein levels (cytoplasm/nucleus), co-immunoprecipitation (GSK3β–GID interaction, MAEA–β-Catenin), Wnt stimulation assay","journal":"Genes to cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, KD rescue, subcellular fractionation, Wnt stimulation) in a single lab establishing pathway position","pmids":["41258755"],"is_preprint":false}],"current_model":"RMND5A is a RING domain-containing subunit of the mammalian CTLH E3 ubiquitin ligase complex (homologous to yeast Gid2/Rmd5 in the GID complex) that, together with MAEA, constitutes the catalytic core mediating both K48- and K63-linked poly-ubiquitin chain formation; it targets substrates including muskelin (for proteasomal degradation), c-Raf/A-Raf/B-Raf (degradation), PKM2 and LDHA (non-degradative activity-modulating ubiquitination to inhibit glycolysis), and β-Catenin (GSK3β-regulated degradation independent of βTrCP), and also functions in neural stem cell self-renewal by modulating Wnt and mTOR signaling."},"narrative":{"mechanistic_narrative":"RMND5A is a RING domain-containing catalytic subunit of the mammalian CTLH E3 ubiquitin ligase complex (the metazoan ortholog of the yeast GID complex), where it assembles with RanBPM, ARMC8, muskelin, WDR26, GID4, and MAEA into a multi-subunit ligase [PMID:17467196, PMID:31285494]. Catalytic competence requires the RING module: in the yeast ortholog Gid2/Rmd5 the degenerate RING finger confers E3 ligase activity and pairs with a second RING subunit (Gid9/Fyv10), both being needed for substrate polyubiquitination and degradation [PMID:18508925, PMID:22044534], a configuration recapitulated in the human complex where RMND5A and MAEA are mutually stabilizing and together support in vitro ubiquitination using UBE2D-family E2 enzymes to build both K48- and K63-linked chains [PMID:31285494]. Through this activity RMND5A directs degradative ubiquitination of substrates including muskelin [PMID:31285494] and the Raf family kinases c-Raf, A-Raf, and B-Raf, with RMND5A loss enhancing proliferation and tumor growth [PMID:30795516], as well as non-degradative ubiquitination of the glycolytic enzymes PKM2 and LDHA, restraining their activity and limiting glycolytic flux [PMID:34383978]. RMND5A also positions the complex within Wnt signaling by mediating GSK3β-regulated, βTrCP-independent degradation of β-Catenin [PMID:41258755], and is required for neural stem/precursor cell self-renewal by modulating Wnt and mTOR signaling [PMID:40377017]. Beyond ubiquitin-pathway roles, RMND5A is itself a target of microRNA regulation (miR-138) that feeds back on Exportin-5 stability and pre-miRNA export [PMID:24057215], and its levels influence cell migration through effects on HDAC6-dependent α-tubulin acetylation [PMID:28668087].","teleology":[{"year":2007,"claim":"Established that the human protein p44CTLH/RMND5A is a constituent subunit of a defined multiprotein CTLH complex, framing all subsequent function within a complex rather than as a standalone protein.","evidence":"Tandem MS, Co-IP, and in vitro pull-down with bacterially expressed Twa1 in mammalian cells","pmids":["17467196"],"confidence":"Medium","gaps":["No catalytic or functional readout assigned specifically to RMND5A","Complex's enzymatic activity not yet demonstrated"]},{"year":2008,"claim":"Identified the catalytic basis of the complex by showing the yeast ortholog's degenerate RING finger confers E3 ligase activity required for FBPase degradation, defining RMND5A's likely role as the ligase module.","evidence":"In vitro ubiquitination with GST-Gid2, RING-domain mutagenesis, and in vivo degradation assay in yeast","pmids":["18508925"],"confidence":"High","gaps":["Demonstrated in yeast ortholog, not human RMND5A","Cognate E2 enzyme not identified"]},{"year":2011,"claim":"Showed the ligase requires two cooperating RING subunits (Gid2/Rmd5 and Gid9/Fyv10), establishing a dual-RING catalytic architecture for substrate ubiquitination.","evidence":"Co-IP, RING-domain mutagenesis, and in vivo ubiquitination/degradation assays in yeast","pmids":["22044534"],"confidence":"High","gaps":["Yeast system; human MAEA partnership not yet shown","Structural basis of RING-RING cooperation unresolved"]},{"year":2012,"claim":"Mapped subunit-subunit contacts within the complex, defining which domains (LisH, CTLH, SPRY) mediate assembly and positioning the RING subunit within the topology.","evidence":"Domain deletion/point mutation followed by Co-IP in yeast","pmids":["22645139"],"confidence":"Medium","gaps":["No high-resolution structure","Human complex topology inferred from yeast"]},{"year":2013,"claim":"Revealed that RMND5A is itself under post-transcriptional control by miR-138 and that its abundance feeds back on Exportin-5 stability and miRNA export, linking RMND5A levels to RNA trafficking and migration.","evidence":"miR-138 overexpression, Western blot, pre-miRNA export assay, and wound-healing assay in HeLa cells","pmids":["24057215"],"confidence":"Medium","gaps":["Whether RMND5A acts on Exportin-5 via ubiquitination not shown","Mechanism connecting RMND5A to migration undefined"]},{"year":2017,"claim":"Connected the CTLH complex to cytoskeletal regulation by showing RMND5A/muskelin loss lowers HDAC6-dependent α-tubulin acetylation and restricts migration.","evidence":"Stable knockdown, Western blot for acetylated α-tubulin, migration assay, and Co-IP","pmids":["28668087"],"confidence":"Medium","gaps":["No direct ubiquitination substrate linking RMND5A to HDAC6 activity","Causal chain to migration indirect"]},{"year":2019,"claim":"Provided definitive reconstitution of human RMND5A as an active E3 ligase, showing it requires MAEA, uses UBE2D-family E2s, builds both K48 and K63 chains, and degrades muskelin via the proteasome.","evidence":"Co-IP, in vitro ubiquitination with recombinant RMND5A, RMND5A KO cells, linkage-specific analysis, proteasome inhibition","pmids":["31285494"],"confidence":"High","gaps":["Determinants of substrate selection not defined","Functional difference between K48 and K63 products on substrates unresolved"]},{"year":2019,"claim":"Identified Raf family kinases as physiological CTLH substrates, linking RMND5A-mediated degradation to control of proliferation and tumor growth.","evidence":"RMND5A depletion, Western blot for Raf isoforms, ubiquitination and proliferation assays, mouse tumor model","pmids":["30795516"],"confidence":"Medium","gaps":["Direct ubiquitination of Raf by recombinant RMND5A not reconstituted","Substrate recognition determinant for Raf unknown"]},{"year":2021,"claim":"Uncovered a non-degradative ubiquitination function by identifying PKM2 and LDHA as RMND5A-dependent substrates whose activity (not abundance) is restrained, coupling the complex to glycolytic control.","evidence":"Global proteomics and diGLY ubiquitinome profiling, RMND5A/RanBPM depletion, enzyme activity and glycolysis assays","pmids":["34383978"],"confidence":"High","gaps":["Ubiquitin linkage type on PKM2/LDHA not defined","Mechanism by which ubiquitination modulates enzyme activity unclear"]},{"year":2021,"claim":"Reported RMND5A as a migration-promoting factor in pancreatic cancer cells under miR-590-5p control, contrasting with earlier migration-restricting observations.","evidence":"Wound-healing assay, RMND5A and miR-590-5p overexpression, Western blot in PAAD lines","pmids":["34079591"],"confidence":"Low","gaps":["No direct molecular mechanism for RMND5A-driven migration","Apparent contradiction with migration-restricting role in HeLa not reconciled"]},{"year":2022,"claim":"Extended the substrate repertoire to the oncopeptide MBOP, degraded by RMND5A/MAEA in colorectal cancer.","evidence":"Co-IP, Western blot, proteasome inhibition in colorectal cancer cells","pmids":["35565466"],"confidence":"Low","gaps":["Single Co-IP without reconstituted ubiquitination","Direct vs indirect substrate relationship not established"]},{"year":2024,"claim":"Linked RMND5A levels in endothelial cells to angiogenic signaling, with RMND5A overexpression suppressing ERK/NF-κB activation and being downregulated by tumor exosomal miR-21.","evidence":"Lentiviral overexpression, MTT, migration, tube formation, Western blot, exosome co-culture","pmids":["38229133"],"confidence":"Low","gaps":["No substrate identified linking RMND5A to ERK/NF-κB","Mechanism of pathway regulation undefined"]},{"year":2025,"claim":"Placed RMND5A within β-Catenin turnover, showing the GID complex (MAEA/RMND5A) degrades β-Catenin in a GSK3β-regulated, βTrCP-independent manner that Wnt stimulation reverses.","evidence":"GSK3β and MAEA/RMND5A knockdown, β-Catenin ubiquitination/fractionation, Co-IP of GSK3β-GID and MAEA-β-Catenin, Wnt stimulation","pmids":["41258755"],"confidence":"Medium","gaps":["Direct β-Catenin ubiquitination by recombinant RMND5A not reconstituted","How GSK3β recruits the complex to β-Catenin not detailed"]},{"year":2025,"claim":"Demonstrated a developmental role for RMND5A in human neural stem/precursor self-renewal through modulation of Wnt and mTOR signaling.","evidence":"CRISPR/Cas9 screen, shRNA knockdown, proliferation/differentiation assays, Wnt and mTOR pathway analysis in hNS/PCs","pmids":["40377017"],"confidence":"Medium","gaps":["Specific ubiquitination substrate in NS/PCs not identified","Direct connection between RMND5A ligase activity and mTOR not shown"]},{"year":null,"claim":"How RMND5A selects its diverse degradative versus non-degradative substrates, and the structural rules governing the RMND5A-MAEA dual-RING catalytic core, remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No high-resolution structure of the human catalytic module","No unified substrate-recognition determinant defined","Reconciliation of context-dependent migration phenotypes lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[1,2,6]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[6,7,8,13]},{"term_id":"GO:0031386","term_label":"protein tag activity","supporting_discovery_ids":[6]}],"localization":[],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[6,7,13]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[7,12,13]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[8]}],"complexes":["CTLH complex","GID complex"],"partners":["MAEA","RANBP9","ARMC8","MKLN1","WDR26","GID4"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9H871","full_name":"E3 ubiquitin-protein transferase RMND5A","aliases":["P44CTLH","Protein RMD5 homolog A"],"length_aa":391,"mass_kda":44.0,"function":"Core component of the CTLH E3 ubiquitin-protein ligase complex that selectively accepts ubiquitin from UBE2H and mediates ubiquitination and subsequent proteasomal degradation of the transcription factor HBP1. MAEA and RMND5A are both required for catalytic activity of the CTLH E3 ubiquitin-protein ligase complex (PubMed:29911972). Catalytic activity of the complex is required for normal cell proliferation (PubMed:29911972). The CTLH E3 ubiquitin-protein ligase complex is not required for the degradation of enzymes involved in gluconeogenesis, such as FBP1 (PubMed:29911972)","subcellular_location":"Nucleus, nucleoplasm; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q9H871/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RMND5A","classification":"Not Classified","n_dependent_lines":467,"n_total_lines":1208,"dependency_fraction":0.38658940397350994},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"RANBP10","stoichiometry":4.0},{"gene":"RANBP9","stoichiometry":4.0}],"url":"https://opencell.sf.czbiohub.org/search/RMND5A","total_profiled":1310},"omim":[{"mim_id":"618964","title":"REQUIRED FOR MEIOTIC NUCLEAR DIVISION 5 HOMOLOG A; RMND5A","url":"https://www.omim.org/entry/618964"},{"mim_id":"617699","title":"GID COMPLEX, SUBUNIT 4; GID4","url":"https://www.omim.org/entry/617699"},{"mim_id":"613394","title":"MICRO RNA 138-1; MIR138-1","url":"https://www.omim.org/entry/613394"},{"mim_id":"607845","title":"EXPORTIN 5; XPO5","url":"https://www.omim.org/entry/607845"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RMND5A"},"hgnc":{"alias_symbol":["FLJ13910","RMD5","GID2","GID2A","p44CTLH"],"prev_symbol":[]},"alphafold":{"accession":"Q9H871","domains":[{"cath_id":"-","chopping":"147-273","consensus_level":"high","plddt":94.3239,"start":147,"end":273},{"cath_id":"3.30.40.10","chopping":"334-387","consensus_level":"high","plddt":91.0485,"start":334,"end":387},{"cath_id":"1.10.287","chopping":"1-47_54-93","consensus_level":"medium","plddt":91.8344,"start":1,"end":93}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H871","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H871-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9H871-F1-predicted_aligned_error_v6.png","plddt_mean":89.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RMND5A","jax_strain_url":"https://www.jax.org/strain/search?query=RMND5A"},"sequence":{"accession":"Q9H871","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9H871.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9H871/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9H871"}},"corpus_meta":[{"pmid":"17965273","id":"PMC_17965273","title":"The GID1-mediated gibberellin perception mechanism is conserved in the Lycophyte Selaginella moellendorffii but not in the Bryophyte Physcomitrella patens.","date":"2007","source":"The Plant cell","url":"https://pubmed.ncbi.nlm.nih.gov/17965273","citation_count":148,"is_preprint":false},{"pmid":"18508925","id":"PMC_18508925","title":"The yeast GID complex, a novel ubiquitin ligase (E3) involved in the regulation of carbohydrate metabolism.","date":"2008","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/18508925","citation_count":138,"is_preprint":false},{"pmid":"11997455","id":"PMC_11997455","title":"Rescue of a pathogenic Marek's disease virus with overlapping cosmid DNAs: use of a pp38 mutant to validate the technology for the study of gene function.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of 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unique region in the Marek's disease virus genome occurs concomitantly with attenuation but is not sufficient to cause attenuation.","date":"2004","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/14694105","citation_count":37,"is_preprint":false},{"pmid":"19941987","id":"PMC_19941987","title":"Comparative evaluation of vaccine efficacy of recombinant Marek's disease virus vaccine lacking Meq oncogene in commercial chickens.","date":"2009","source":"Vaccine","url":"https://pubmed.ncbi.nlm.nih.gov/19941987","citation_count":34,"is_preprint":false},{"pmid":"34383978","id":"PMC_34383978","title":"Proteomic analysis of ubiquitination substrates reveals a CTLH E3 ligase complex-dependent regulation of glycolysis.","date":"2021","source":"FASEB journal : official publication of the Federation of American Societies for Experimental 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Series C, Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/16544576","citation_count":4,"is_preprint":false},{"pmid":"23703623","id":"PMC_23703623","title":"Visualization of Marek's disease virus in vitro using enhanced green fluorescent protein fused with US10.","date":"2013","source":"Virus genes","url":"https://pubmed.ncbi.nlm.nih.gov/23703623","citation_count":4,"is_preprint":false},{"pmid":"31657656","id":"PMC_31657656","title":"Effect of low frequency magnetic field on efficiency of chromosome break repair.","date":"2019","source":"Electromagnetic biology and medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31657656","citation_count":3,"is_preprint":false},{"pmid":"39861015","id":"PMC_39861015","title":"Protection Conferred by Gallid Alphaherpesvirus 2 Vaccines Against Immunosuppression Induced by Very Virulent Plus (vv+) Marek's Disease Virus Strains in Commercial Meat Type Chickens.","date":"2025","source":"Pathogens (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/39861015","citation_count":3,"is_preprint":false},{"pmid":"40883813","id":"PMC_40883813","title":"RANBP9 and RANBP10 cooperate in regulating non-small cell lung cancer proliferation.","date":"2025","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/40883813","citation_count":2,"is_preprint":false},{"pmid":"39899476","id":"PMC_39899476","title":"Marek's disease virus-1 unique gene LORF1 is involved in viral replication and MDV-1/Md5-induced atrophy of the bursa of Fabricius.","date":"2025","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/39899476","citation_count":2,"is_preprint":false},{"pmid":"40377017","id":"PMC_40377017","title":"E3 ubiquitin ligase RMND5A maintains the self-renewal state of human neural stem/precursor cells by regulating Wnt and mTOR signaling pathways.","date":"2025","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/40377017","citation_count":1,"is_preprint":false},{"pmid":"41258755","id":"PMC_41258755","title":"GSK3β Regulates a Novel β-Catenin Degradation Pathway via the GID Complex in Wnt Signaling.","date":"2025","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/41258755","citation_count":1,"is_preprint":false},{"pmid":"41284154","id":"PMC_41284154","title":"RMND5A upregulation via IGF2BP3-mediated m6A RNA modification enhances malignant traits and immune evasion in laryngeal squamous cell carcinoma cells.","date":"2025","source":"Human cell","url":"https://pubmed.ncbi.nlm.nih.gov/41284154","citation_count":0,"is_preprint":false},{"pmid":"41050213","id":"PMC_41050213","title":"Identification of novel high-risk genes in gastric cancer through single-cell RNA sequencing, eQTL Mendelian randomization, and in vitro validation.","date":"2025","source":"Global medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/41050213","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":29203,"output_tokens":3806,"usd":0.072349,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11451,"output_tokens":4385,"usd":0.08344,"stage2_stop_reason":"end_turn"},"total_usd":0.155789,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"RMND5A (p44CTLH) was identified as a component of the mammalian CTLH complex, along with RanBPM, Muskelin, p48EMLP, ARMC8α, and ARMC8β. Each component contains LisH/CTLH motifs. Co-immunoprecipitation and pull-down assays with bacterially-expressed Twa1 confirmed in vivo and in vitro associations of all complex members.\",\n      \"method\": \"Tandem MS immunoprecipitation, co-immunoprecipitation (Co-IP), and in vitro pull-down assay with bacterially-expressed Twa1\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple orthogonal methods (MS, Co-IP, pull-down) in a single lab; complex membership established but no functional readout specifically for RMND5A\",\n      \"pmids\": [\"17467196\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"The yeast ortholog Gid2/Rmd5 contains a degenerate RING finger domain and provides E3 ubiquitin ligase activity to the Gid complex; mutation of the degenerate RING domain abolishes fructose-1,6-bisphosphatase (FBPase) polyubiquitination and degradation in vivo, and heterologous GST-Gid2 expression leads to polyubiquitination in vitro.\",\n      \"method\": \"In vitro ubiquitination assay (GST-Gid2 expression), RING domain mutagenesis, in vivo degradation assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro ubiquitination assay plus active-site mutagenesis plus in vivo degradation phenotype in yeast ortholog; multiple orthogonal methods in one study\",\n      \"pmids\": [\"18508925\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Yeast Gid2/Rmd5 physically interacts with a second RING finger subunit Gid9/Fyv10 within the Gid complex; mutation of Gid9's RING finger abolishes polyubiquitylation and degradation of three gluconeogenic enzymes, indicating that both RING subunits are required for full E3 ligase activity.\",\n      \"method\": \"Co-immunoprecipitation, RING domain mutagenesis, in vivo ubiquitination/degradation assay\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — reciprocal interaction confirmed, active-site mutagenesis with functional readout in yeast ortholog, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"22044534\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Domain-deletion and point-mutation analysis of yeast Gid complex subunits (including Gid2/Rmd5) revealed the topology of subunit interactions; LisH, CTLH, and SPRY domains in individual Gid proteins mediate specific subunit–subunit contacts within the E3 ubiquitin ligase complex.\",\n      \"method\": \"Domain deletion/mutagenesis followed by co-immunoprecipitation to map subunit interactions\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic domain mutagenesis with Co-IP readout in yeast ortholog, single lab\",\n      \"pmids\": [\"22645139\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RMND5A was identified as a direct target of miR-138 in HeLa cells; overexpression of miR-138 downregulated RMND5A protein, which in turn reduced Exportin-5 stability and decreased pre-miRNA nuclear export, and also significantly inhibited HeLa cell migration.\",\n      \"method\": \"miR-138 overexpression, Western blot (RMND5A and Exportin-5 levels), pre-miRNA export assay, wound-healing migration assay\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — multiple functional readouts (protein stability, RNA export, migration) in a single lab; direct experimental validation of miR-138→RMND5A→Exportin-5 pathway\",\n      \"pmids\": [\"24057215\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Downregulation of RMND5A (along with muskelin) decreased acetylation of the HDAC6 substrate α-tubulin, demonstrating that the CTLH complex (via RMND5A) contributes to inhibition of HDAC6 deacetylase activity and consequently restricts cell migration.\",\n      \"method\": \"Stable knockdown cell lines, Western blot for acetylated α-tubulin, wound-healing migration assay, co-immunoprecipitation\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — functional knockdown with two orthogonal readouts (biochemical activity and cell migration), single lab\",\n      \"pmids\": [\"28668087\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The human CTLH complex immunoprecipitated from cells comprises RanBPM, ARMC8α/β, muskelin, WDR26, GID4, RMND5A, and MAEA. RMND5A and MAEA protein levels are interdependent. In vitro ubiquitination assays showed E3 ligase activity dependent on both RMND5A and MAEA. Recombinant RMND5A mediates K48 and K63 poly-ubiquitin chains and pairs with UBE2D1, UBE2D2, and UBE2D3 E2 enzymes. Muskelin ubiquitination is RMND5A-dependent, and muskelin protein levels increase in RMND5A KO cells in a proteasome-dependent manner.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay with recombinant RMND5A, RMND5A knockout cells, ubiquitin linkage-specific analysis, proteasome inhibitor treatment\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution with recombinant protein, active-site-dependent assay, KO cellular validation, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"31285494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"The CTLH complex promotes c-Raf ubiquitination and proteasome-dependent degradation; depletion of RMND5A (or RanBPM) results in enhanced cell proliferation and tumor growth. A-Raf and B-Raf levels are also regulated by the complex, indicating a common Raf family regulation.\",\n      \"method\": \"RMND5A depletion, Western blot for c-Raf/A-Raf/B-Raf levels, ubiquitination assay, proliferation assay, mouse tumor model\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO/KD with multiple functional and biochemical readouts, in vivo mouse model, single lab\",\n      \"pmids\": [\"30795516\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Proteomic and diGLY-enriched ubiquitinome analysis in CTLH-depleted HeLa cells identified glycolysis enzymes PKM2 and LDHA as RMND5A-dependent ubiquitination substrates. Reduced polyubiquitination of PKM2 and LDHA was validated in RMND5A-depleted cells; their enzymatic activities were increased without changes in protein levels, and RanBPM-deficient cells exhibited enhanced glycolysis, uncovering a non-degradative ubiquitination role for RMND5A/CTLH.\",\n      \"method\": \"Mass spectrometry-based global proteomics, diGLY-enriched ubiquitinome profiling, RanBPM/RMND5A depletion, enzymatic activity assays (PKM2, LDHA), glycolysis metabolic assay\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal mass spectrometry approaches plus direct biochemical validation of substrate ubiquitination and enzyme activity, single lab with comprehensive methodology\",\n      \"pmids\": [\"34383978\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RMND5A and MAEA (GID complex RING subunits) mediate ubiquitin-proteasome-dependent degradation of the oncopeptide MBOP encoded by LINC01234 in colorectal cancer cells, as shown by immunoprecipitation experiments.\",\n      \"method\": \"Co-immunoprecipitation, Western blot, proteasome inhibitor treatment\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP experiment identifying RMND5A as one of two E3 ligases for a specific substrate, single lab, limited mechanistic follow-up\",\n      \"pmids\": [\"35565466\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RMND5A overexpression significantly increased cell migration in pancreatic adenocarcinoma cell lines (AsPC-1 and PANC-1), and miR-590-5p-mediated downregulation of RMND5A decreased this migratory ability, placing RMND5A upstream of migration-promoting signaling in PAAD cells.\",\n      \"method\": \"Wound-healing migration assay, RMND5A overexpression, miR-590-5p overexpression, Western blot\",\n      \"journal\": \"Oncology letters\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, phenotypic rescue experiment but no direct molecular mechanism identified for how RMND5A promotes migration\",\n      \"pmids\": [\"34079591\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Overexpression of RMND5A in HUVECs reduced proliferation, migration, and tube formation by inhibiting ERK and NF-κB pathway activation. OSCC-derived exosomal miR-21 suppresses RMND5A expression in endothelial cells to promote angiogenesis.\",\n      \"method\": \"Lentiviral RMND5A overexpression, MTT assay, wound-healing assay, tube formation assay, Western blot (ERK/NF-κB), exosome co-culture\",\n      \"journal\": \"BMC oral health\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, overexpression-based phenotypic assay without direct substrate identification or molecular mechanism for ERK/NF-κB regulation by RMND5A\",\n      \"pmids\": [\"38229133\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RMND5A knockdown in human neural stem/precursor cells (hNS/PCs) decreased proliferation and promoted neuronal differentiation, associated with activation of Wnt signaling and suppression of mTOR signaling, identifying RMND5A as required for hNS/PC self-renewal.\",\n      \"method\": \"CRISPR/Cas9 knockout screen, shRNA knockdown, proliferation and differentiation assays, Western blot/pathway analysis (Wnt, mTOR)\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — CRISPR screen plus targeted KD with multiple functional readouts (proliferation, differentiation, pathway activation), single lab\",\n      \"pmids\": [\"40377017\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The GID complex (via MAEA and RMND5A subunits) ubiquitinates and degrades β-Catenin independently of βTrCP when GSK3β is suppressed. Wnt stimulation promotes GSK3β–GID complex interaction, disrupting the MAEA–β-Catenin association and thereby stabilizing β-Catenin. Knockdown of MAEA and RMND5A rescued β-Catenin levels in GSK3β-knockdown cells.\",\n      \"method\": \"GSK3β knockdown, MAEA/RMND5A knockdown, Western blot for β-Catenin ubiquitination and protein levels (cytoplasm/nucleus), co-immunoprecipitation (GSK3β–GID interaction, MAEA–β-Catenin), Wnt stimulation assay\",\n      \"journal\": \"Genes to cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (Co-IP, KD rescue, subcellular fractionation, Wnt stimulation) in a single lab establishing pathway position\",\n      \"pmids\": [\"41258755\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RMND5A is a RING domain-containing subunit of the mammalian CTLH E3 ubiquitin ligase complex (homologous to yeast Gid2/Rmd5 in the GID complex) that, together with MAEA, constitutes the catalytic core mediating both K48- and K63-linked poly-ubiquitin chain formation; it targets substrates including muskelin (for proteasomal degradation), c-Raf/A-Raf/B-Raf (degradation), PKM2 and LDHA (non-degradative activity-modulating ubiquitination to inhibit glycolysis), and β-Catenin (GSK3β-regulated degradation independent of βTrCP), and also functions in neural stem cell self-renewal by modulating Wnt and mTOR signaling.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RMND5A is a RING domain-containing catalytic subunit of the mammalian CTLH E3 ubiquitin ligase complex (the metazoan ortholog of the yeast GID complex), where it assembles with RanBPM, ARMC8, muskelin, WDR26, GID4, and MAEA into a multi-subunit ligase [#0, #6]. Catalytic competence requires the RING module: in the yeast ortholog Gid2/Rmd5 the degenerate RING finger confers E3 ligase activity and pairs with a second RING subunit (Gid9/Fyv10), both being needed for substrate polyubiquitination and degradation [#1, #2], a configuration recapitulated in the human complex where RMND5A and MAEA are mutually stabilizing and together support in vitro ubiquitination using UBE2D-family E2 enzymes to build both K48- and K63-linked chains [#6]. Through this activity RMND5A directs degradative ubiquitination of substrates including muskelin [#6] and the Raf family kinases c-Raf, A-Raf, and B-Raf, with RMND5A loss enhancing proliferation and tumor growth [#7], as well as non-degradative ubiquitination of the glycolytic enzymes PKM2 and LDHA, restraining their activity and limiting glycolytic flux [#8]. RMND5A also positions the complex within Wnt signaling by mediating GSK3\\u03b2-regulated, \\u03b2TrCP-independent degradation of \\u03b2-Catenin [#13], and is required for neural stem/precursor cell self-renewal by modulating Wnt and mTOR signaling [#12]. Beyond ubiquitin-pathway roles, RMND5A is itself a target of microRNA regulation (miR-138) that feeds back on Exportin-5 stability and pre-miRNA export [#4], and its levels influence cell migration through effects on HDAC6-dependent \\u03b1-tubulin acetylation [#5].\"\n,\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established that the human protein p44CTLH/RMND5A is a constituent subunit of a defined multiprotein CTLH complex, framing all subsequent function within a complex rather than as a standalone protein.\",\n      \"evidence\": \"Tandem MS, Co-IP, and in vitro pull-down with bacterially expressed Twa1 in mammalian cells\",\n      \"pmids\": [\"17467196\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No catalytic or functional readout assigned specifically to RMND5A\", \"Complex's enzymatic activity not yet demonstrated\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identified the catalytic basis of the complex by showing the yeast ortholog's degenerate RING finger confers E3 ligase activity required for FBPase degradation, defining RMND5A's likely role as the ligase module.\",\n      \"evidence\": \"In vitro ubiquitination with GST-Gid2, RING-domain mutagenesis, and in vivo degradation assay in yeast\",\n      \"pmids\": [\"18508925\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Demonstrated in yeast ortholog, not human RMND5A\", \"Cognate E2 enzyme not identified\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed the ligase requires two cooperating RING subunits (Gid2/Rmd5 and Gid9/Fyv10), establishing a dual-RING catalytic architecture for substrate ubiquitination.\",\n      \"evidence\": \"Co-IP, RING-domain mutagenesis, and in vivo ubiquitination/degradation assays in yeast\",\n      \"pmids\": [\"22044534\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Yeast system; human MAEA partnership not yet shown\", \"Structural basis of RING-RING cooperation unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Mapped subunit-subunit contacts within the complex, defining which domains (LisH, CTLH, SPRY) mediate assembly and positioning the RING subunit within the topology.\",\n      \"evidence\": \"Domain deletion/point mutation followed by Co-IP in yeast\",\n      \"pmids\": [\"22645139\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution structure\", \"Human complex topology inferred from yeast\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Revealed that RMND5A is itself under post-transcriptional control by miR-138 and that its abundance feeds back on Exportin-5 stability and miRNA export, linking RMND5A levels to RNA trafficking and migration.\",\n      \"evidence\": \"miR-138 overexpression, Western blot, pre-miRNA export assay, and wound-healing assay in HeLa cells\",\n      \"pmids\": [\"24057215\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether RMND5A acts on Exportin-5 via ubiquitination not shown\", \"Mechanism connecting RMND5A to migration undefined\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected the CTLH complex to cytoskeletal regulation by showing RMND5A/muskelin loss lowers HDAC6-dependent \\u03b1-tubulin acetylation and restricts migration.\",\n      \"evidence\": \"Stable knockdown, Western blot for acetylated \\u03b1-tubulin, migration assay, and Co-IP\",\n      \"pmids\": [\"28668087\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct ubiquitination substrate linking RMND5A to HDAC6 activity\", \"Causal chain to migration indirect\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Provided definitive reconstitution of human RMND5A as an active E3 ligase, showing it requires MAEA, uses UBE2D-family E2s, builds both K48 and K63 chains, and degrades muskelin via the proteasome.\",\n      \"evidence\": \"Co-IP, in vitro ubiquitination with recombinant RMND5A, RMND5A KO cells, linkage-specific analysis, proteasome inhibition\",\n      \"pmids\": [\"31285494\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Determinants of substrate selection not defined\", \"Functional difference between K48 and K63 products on substrates unresolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified Raf family kinases as physiological CTLH substrates, linking RMND5A-mediated degradation to control of proliferation and tumor growth.\",\n      \"evidence\": \"RMND5A depletion, Western blot for Raf isoforms, ubiquitination and proliferation assays, mouse tumor model\",\n      \"pmids\": [\"30795516\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitination of Raf by recombinant RMND5A not reconstituted\", \"Substrate recognition determinant for Raf unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Uncovered a non-degradative ubiquitination function by identifying PKM2 and LDHA as RMND5A-dependent substrates whose activity (not abundance) is restrained, coupling the complex to glycolytic control.\",\n      \"evidence\": \"Global proteomics and diGLY ubiquitinome profiling, RMND5A/RanBPM depletion, enzyme activity and glycolysis assays\",\n      \"pmids\": [\"34383978\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitin linkage type on PKM2/LDHA not defined\", \"Mechanism by which ubiquitination modulates enzyme activity unclear\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Reported RMND5A as a migration-promoting factor in pancreatic cancer cells under miR-590-5p control, contrasting with earlier migration-restricting observations.\",\n      \"evidence\": \"Wound-healing assay, RMND5A and miR-590-5p overexpression, Western blot in PAAD lines\",\n      \"pmids\": [\"34079591\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct molecular mechanism for RMND5A-driven migration\", \"Apparent contradiction with migration-restricting role in HeLa not reconciled\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Extended the substrate repertoire to the oncopeptide MBOP, degraded by RMND5A/MAEA in colorectal cancer.\",\n      \"evidence\": \"Co-IP, Western blot, proteasome inhibition in colorectal cancer cells\",\n      \"pmids\": [\"35565466\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP without reconstituted ubiquitination\", \"Direct vs indirect substrate relationship not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked RMND5A levels in endothelial cells to angiogenic signaling, with RMND5A overexpression suppressing ERK/NF-\\u03baB activation and being downregulated by tumor exosomal miR-21.\",\n      \"evidence\": \"Lentiviral overexpression, MTT, migration, tube formation, Western blot, exosome co-culture\",\n      \"pmids\": [\"38229133\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No substrate identified linking RMND5A to ERK/NF-\\u03baB\", \"Mechanism of pathway regulation undefined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Placed RMND5A within \\u03b2-Catenin turnover, showing the GID complex (MAEA/RMND5A) degrades \\u03b2-Catenin in a GSK3\\u03b2-regulated, \\u03b2TrCP-independent manner that Wnt stimulation reverses.\",\n      \"evidence\": \"GSK3\\u03b2 and MAEA/RMND5A knockdown, \\u03b2-Catenin ubiquitination/fractionation, Co-IP of GSK3\\u03b2-GID and MAEA-\\u03b2-Catenin, Wnt stimulation\",\n      \"pmids\": [\"41258755\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct \\u03b2-Catenin ubiquitination by recombinant RMND5A not reconstituted\", \"How GSK3\\u03b2 recruits the complex to \\u03b2-Catenin not detailed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Demonstrated a developmental role for RMND5A in human neural stem/precursor self-renewal through modulation of Wnt and mTOR signaling.\",\n      \"evidence\": \"CRISPR/Cas9 screen, shRNA knockdown, proliferation/differentiation assays, Wnt and mTOR pathway analysis in hNS/PCs\",\n      \"pmids\": [\"40377017\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Specific ubiquitination substrate in NS/PCs not identified\", \"Direct connection between RMND5A ligase activity and mTOR not shown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RMND5A selects its diverse degradative versus non-degradative substrates, and the structural rules governing the RMND5A-MAEA dual-RING catalytic core, remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution structure of the human catalytic module\", \"No unified substrate-recognition determinant defined\", \"Reconciliation of context-dependent migration phenotypes lacking\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [1, 2, 6]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [6, 7, 8, 13]},\n      {\"term_id\": \"GO:0031386\", \"supporting_discovery_ids\": [6]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [6, 7, 13]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [7, 12, 13]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"complexes\": [\"CTLH complex\", \"GID complex\"],\n    \"partners\": [\"MAEA\", \"RANBP9\", \"ARMC8\", \"MKLN1\", \"WDR26\", \"GID4\"],\n    \"other_free_text\": []\n  }\n}\n```","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":5,"faith_pct":80.0}}