{"gene":"RNF10","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2021,"finding":"RNF10 is the E3 ubiquitin ligase responsible for site-specific monoubiquitylation of 40S ribosomal proteins uS3 (RPS3) and uS5 (RPS2) during ribosome-associated quality control (RQC). USP10 is the counteracting deubiquitylase. Prolonged uS3/uS5 ubiquitylation leads to selective 40S (but not 60S) ribosomal protein degradation independent of canonical autophagy. This pathway, termed initiation RQC (iRQC), is triggered when scanning or elongating ribosomes are blocked from progressing past the start codon.","method":"Genetic screens, quantitative proteomics, knockdown/knockout with ribosome degradation readouts, translation reporter assays","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 — reciprocal identification of ligase/deubiquitylase pair with multiple orthogonal methods; independently replicated in companion paper (PMID:34348161)","pmids":["34469731"],"is_preprint":false},{"year":2021,"finding":"RNF10 monoubiquitinates RPS2/uS5 and RPS3/uS3 on ribosomes stalled in translation; overexpression of RNF10 phenocopies USP10 knockout by increasing 40S subunit degradation. PAR-CLIP showed RNF10 crosslinks to mRNAs, tRNAs, and 18S rRNA, indicating direct recruitment to stalled ribosomes. ZNF598-independent translation initiation and elongation impairment also contributes to RNF10-mediated ubiquitination.","method":"Overexpression/knockout cell lines, PAR-CLIP, ribosome fractionation, ubiquitination assays","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 1-2 — direct biochemical identification of substrates with PAR-CLIP crosslinking and ribosome fractionation; independently corroborated by PMID:34469731","pmids":["34348161"],"is_preprint":false},{"year":2025,"finding":"Mammalian 18S nonfunctional rRNA decay (NRD) proceeds through a GCN2-RNF10-RIOK3 axis: nonfunctional 18S rRNA induces translational arrest at start sites, activating GCN2 (integrated stress response), which limits translation initiation; RNF10-mediated ubiquitination of 40S proteins then promotes 40S ribosomal protein turnover and 18S rRNA decay, with RIOK3 binding ubiquitinated 40S subunits to facilitate 18S rRNA degradation.","method":"Genome-wide CRISPR genetic interaction screens, selective ribosome profiling, biochemical ISR activation assays, 18S rRNA decay assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1-2 — genome-wide epistasis screen plus biochemical validation; published in peer-reviewed journal with multiple orthogonal methods","pmids":["39947182"],"is_preprint":false},{"year":2025,"finding":"RIOK3 specifically recognizes RNF10-ubiquitylated 40S ribosomes through a unique ubiquitin-interacting motif (visualized by cryo-EM), and mediates progressive 3'-to-5' decay of 18S rRNA in the ubiquitylated 40S subunit. Starvation induces selective depletion of 40S ribosomes via RNF10 ubiquitylation followed by RIOK3-dependent degradation.","method":"Cryo-EM structure of RIOK3–ubiquitylated 40S complex, genetic knockouts, ribosome degradation assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — cryo-EM structural validation of ubiquitin-interacting motif plus functional genetics; independent parallel study corroborates (PMID:39947182)","pmids":["39947183"],"is_preprint":false},{"year":2025,"finding":"Disruption of 60S biogenesis triggers iRQC activation and 40S decay via RNF10-mediated ubiquitylation of uS3/uS5; depletion of the scanning helicase eIF4A1 impairs 40S ubiquitylation, indicating mRNA engagement is required for iRQC. Amino acid starvation also stimulates iRQC-dependent 40S decay. RIOK3 interacts with ubiquitylated 40S subunits to mediate degradation, and both RNF10 and RIOK3 protein levels increase upon iRQC activation (feedforward mechanism).","method":"Genetic knockdowns of 60S/40S biogenesis factors, eIF4A1 depletion, amino acid starvation assays, co-immunoprecipitation of RIOK3 with ubiquitylated 40S","journal":"Cell reports","confidence":"High","confidence_rationale":"Tier 2 — multiple genetic perturbations with defined phenotypic readouts; corroborated by PMID:39947182 and PMID:39947183","pmids":["40022732"],"is_preprint":false},{"year":2024,"finding":"RNF10-mediated monoubiquitination of RPS3/uS3 antagonizes ribosomal half-mer formation by promoting dissociation of 40S subunits from ribosomes stalled during both translation elongation and aberrant initiation. RNF10 protein levels are coupled to 40S subunit abundance: knockdown of RPS proteins leads to proteasomal degradation of RNF10, whereas knockdown of RPL proteins causes accumulation of stalled initiating 40S subunits and increased RNF10 levels.","method":"Ribosome fractionation, half-mer analysis, RPS/RPL knockdowns, proteasome inhibition, ubiquitination assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal approaches showing mechanistic link between RNF10 activity and ribosomal subunit stoichiometry","pmids":["39609413"],"is_preprint":false},{"year":2025,"finding":"The E3 ubiquitin ligase LTN1 suppresses RNF10 expression in a manner dependent on LTN1's RING domain, revealing crosstalk between RQC-associated E3 ligases as a mechanism coordinating translational surveillance pathways.","method":"Knockout mouse and human cell lines, western blotting, RING domain mutant analysis","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 — clean genetic knockouts with domain-level mechanistic dissection; single study","pmids":["41451945"],"is_preprint":false},{"year":2016,"finding":"RNF10 is a synaptonuclear messenger enriched at excitatory synapses where it associates with the GluN2A subunit of NMDA receptors. Activation of synaptic GluN2A-containing NMDARs and LTP induction cause RNF10 translocation from dendritic segments and spines to the nucleus via importin-dependent long-distance transport. RNF10 silencing prevents LTP maintenance and LTP-dependent structural modifications of dendritic spines.","method":"Co-immunoprecipitation with GluN2A, live-cell imaging, importin inhibition, siRNA knockdown with LTP electrophysiology and spine morphology readouts","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, direct imaging of translocation, functional loss-of-function with defined electrophysiological and structural phenotypes","pmids":["26977767"],"is_preprint":false},{"year":2019,"finding":"PKC-dependent phosphorylation of RNF10 at Ser31 is required for RNF10 detachment from the NMDA receptor GluN2A subunit and subsequent nuclear trafficking. Preventing Ser31 phosphorylation decreases spine density, neuronal branching, and CREB signaling; mimicking stable Ser31 phosphorylation has opposite effects.","method":"Phosphomimetic/phosphodead mutants, live-cell imaging, dendritic spine morphometry, CREB reporter assays","journal":"Molecular neurobiology","confidence":"High","confidence_rationale":"Tier 2 — site-directed mutagenesis of phosphosite with multiple structural and signaling readouts","pmids":["31069631"],"is_preprint":false},{"year":2008,"finding":"RNF10 binds to a cis-acting element ~160 bp upstream of the MAG transcription start site and acts as a transcriptional activator of the myelin-associated glycoprotein (MAG) gene in Schwann cells. RNF10 knockdown reduces endogenous MAG mRNA and protein; retroviral RNF10 siRNA in Schwann cell–DRG neuron co-cultures inhibits myelin formation.","method":"Yeast one-hybrid screen, luciferase reporter assay, siRNA knockdown, retroviral transduction, Schwann cell–DRG neuron myelination co-culture","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 — yeast one-hybrid identification of cis-element binding, reporter assays, and functional myelination readout with siRNA; multiple orthogonal methods","pmids":["18941509"],"is_preprint":false},{"year":2005,"finding":"RNF10 physically interacts with the transcription factor MEOX2 via a central region of MEOX2 (amino acids 101–185); the RING finger domain of RNF10 is not required for MEOX2 binding. RNF10 co-expression enhances MEOX2-mediated activation of the p21WAF1 promoter.","method":"Yeast two-hybrid screen, in vitro pull-down, co-immunoprecipitation in mammalian cells, deletion mapping, luciferase reporter assay","journal":"Molecular and cellular biochemistry","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP plus in vitro pull-down and domain mapping with functional reporter assay","pmids":["16335786"],"is_preprint":false},{"year":2013,"finding":"RNF10 expression increases upon retinoic acid-induced neuronal differentiation of P19 cells; RNF10 knockdown impairs neuronal differentiation and prevents cell cycle arrest after RA treatment. RNF10 regulates cell cycle exit through upregulation of p21 (but not p27 or p57), and ectopic p21 partially rescues the differentiation defect caused by RNF10 depletion.","method":"siRNA knockdown, BrdU incorporation, flow cytometry cell cycle profiling, neuronal marker western blots, p21 rescue experiment","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 — clean knockdown with specific epistasis rescue by p21; single lab study","pmids":["23526782"],"is_preprint":false},{"year":2013,"finding":"RNF10 is identified as a target of S-nitrosylation; eight ubiquitin E3 ligases including RNF10 were found to be potentially S-nitrosylated, suggesting NO-mediated regulation of RNF10's ubiquitin ligase activity.","method":"High-density protein microarray with S-nitrosylation-specific labeling and affinity capture; mass spectrometry identification of modified cysteines","journal":"Molecular & cellular proteomics","confidence":"Low","confidence_rationale":"Tier 3 — proteome-wide screen identifies RNF10 as S-nitrosylated but functional consequence on ligase activity not directly validated","pmids":["24105792"],"is_preprint":false},{"year":2024,"finding":"In Drosophila, the E3 ligases CNOT4 and RNF10 function upstream of the deubiquitinase OTUD6 to regulate ubiquitination of RPS7/eS7 on the free 40S ribosome, modulating global protein translation and the response to alkylation stress.","method":"Coimmunoprecipitation, enrichment of monoubiquitinated proteins from catalytically inactive OTUD6 Drosophila, genetic epistasis","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis in Drosophila ortholog context with Co-IP validation; single lab study","pmids":["39127721"],"is_preprint":false},{"year":2021,"finding":"RNF10 knockdown in macrophages enhances both NF-κB and IRF3 signaling pathways, leading to increased proinflammatory cytokines and type I interferons, and promoting clearance of Listeria monocytogenes, indicating RNF10 acts as a negative regulator of innate immune signaling in macrophages.","method":"siRNA knockdown in macrophages, NF-κB/IRF3 pathway reporter assays, cytokine/interferon quantification, bacterial clearance assay","journal":"FEBS open bio","confidence":"Medium","confidence_rationale":"Tier 2 — clean knockdown with defined signaling and functional readouts; single lab study","pmids":["33249776"],"is_preprint":false},{"year":2026,"finding":"In the context of vascular calcification in chronic kidney disease, nuclear RNF10 negatively regulates Rbpjk expression in VSMCs through a transcriptional (non-ubiquitin-ligase) mechanism. Proteasome inhibition does not impair RNF10's anticalcific activity. Viral Rbpjk overexpression reverses RNF10's protective effects, while Rbpjk knockdown reduces osteogenic markers, defining an RNF10-Rbpjk regulatory axis.","method":"Rnf10 knock-in rats, RNF10 overexpression in VSMCs, RNA-seq, ChIP-seq, ChIP-qPCR, luciferase reporter assays, gain- and loss-of-function for Rbpjk in vivo and in vitro, proteasome inhibition","journal":"Arteriosclerosis, thrombosis, and vascular biology","confidence":"High","confidence_rationale":"Tier 2 — in vivo knock-in model combined with ChIP-seq, epistasis experiments, and multiple orthogonal validation methods","pmids":["41988714"],"is_preprint":false},{"year":2007,"finding":"RNF10 interacts with the tumor suppressor RASSF1C in a yeast two-hybrid screen confirmed by in vitro pull-down of bacterially expressed proteins, placing RNF10 within a nuclear interactome network that includes hampin/MSL1 and associated chromatin regulators.","method":"Yeast two-hybrid library screen, in vitro pull-down with bacterially expressed proteins","journal":"Biochemical and biophysical research communications","confidence":"Low","confidence_rationale":"Tier 3 — yeast two-hybrid plus in vitro pull-down only; functional consequence not established","pmids":["17335777"],"is_preprint":false},{"year":2011,"finding":"RNF10 interacts with HSV-1 tegument protein VP22; co-expression of VP22 with RNF10 relocates RNF10 from its normal subcellular distribution pattern, indicating VP22 modulates RNF10 localization.","method":"Yeast two-hybrid, fluorescent protein tagging and co-expression imaging","journal":"Archives of virology","confidence":"Low","confidence_rationale":"Tier 3 — yeast two-hybrid and localization imaging only; no direct functional mechanistic follow-up","pmids":["21424732"],"is_preprint":false}],"current_model":"RNF10 is a RING-domain E3 ubiquitin ligase that functions at multiple levels: in the ribosome quality control pathway it monoubiquitylates 40S small subunit proteins uS3/RPS3 and uS5/RPS2 on stalled or initiating ribosomes (iRQC), promoting 40S subunit dissociation and selective 40S degradation via RIOK3-mediated 18S rRNA decay in a GCN2-dependent integrated stress response; in neurons it serves as a calcium/NMDAR-activated synaptonuclear messenger that undergoes PKC-phosphorylation at Ser31, importin-dependent transport to the nucleus, and regulates transcription required for LTP maintenance and dendritic spine morphology; and in non-neuronal cells it acts as a transcriptional regulator (of MAG in Schwann cells and Rbpjk in VSMCs) and interacts with MEOX2 to activate p21-dependent cell cycle exit."},"narrative":{"teleology":[{"year":2005,"claim":"Identification of RNF10 as a physical partner of the homeodomain transcription factor MEOX2 established RNF10's first known protein interaction and linked it to p21-dependent transcriptional regulation, raising the question of whether RNF10 functions beyond canonical E3 ligase activity.","evidence":"Yeast two-hybrid, co-IP, in vitro pull-down, and p21 promoter reporter assay in mammalian cells","pmids":["16335786"],"confidence":"High","gaps":["RING domain dispensability for MEOX2 binding was shown but the RING domain's catalytic role was not tested in this context","endogenous co-expression and physiological relevance not established"]},{"year":2008,"claim":"Discovery that RNF10 directly binds a cis-element upstream of the MAG gene and activates MAG transcription in Schwann cells established RNF10 as a bona fide transcriptional activator controlling peripheral myelination.","evidence":"Yeast one-hybrid screen, luciferase reporters, siRNA knockdown, and Schwann cell–DRG neuron myelination co-culture","pmids":["18941509"],"confidence":"High","gaps":["Mechanism by which a RING-domain protein binds DNA and activates transcription not resolved","relationship between ubiquitin ligase activity and transcriptional function not tested"]},{"year":2013,"claim":"Demonstration that RNF10 is required for retinoic acid-induced neuronal differentiation and cell cycle exit through p21 upregulation connected RNF10's transcriptional co-activator role with MEOX2 to a defined developmental process.","evidence":"siRNA knockdown in P19 cells, cell cycle profiling, p21 rescue experiment","pmids":["23526782"],"confidence":"Medium","gaps":["Study performed in a single cell line (P19)","direct transcriptional mechanism on p21 promoter not dissected","whether RNF10 E3 activity is involved in cell cycle regulation unclear"]},{"year":2016,"claim":"Identification of RNF10 as an activity-dependent synaptonuclear messenger that translocates from NMDAR/GluN2A complexes to the nucleus upon LTP induction revealed a non-canonical signaling function for an E3 ligase in synaptic plasticity.","evidence":"Co-IP with GluN2A, live-cell imaging, importin inhibition, siRNA knockdown with LTP electrophysiology and spine morphology readouts in neurons","pmids":["26977767"],"confidence":"High","gaps":["Nuclear transcriptional targets mediating LTP maintenance not identified","whether ubiquitin ligase activity is required for the synaptic function not tested"]},{"year":2019,"claim":"Pinpointing PKC-dependent phosphorylation of Ser31 as the molecular switch controlling RNF10 detachment from GluN2A and nuclear import resolved the activation mechanism of the synaptonuclear signaling pathway.","evidence":"Phosphomimetic/phosphodead mutants, live-cell imaging, spine morphometry, CREB reporter assays in neurons","pmids":["31069631"],"confidence":"High","gaps":["Identity of the specific PKC isoform responsible not determined","downstream nuclear gene targets of RNF10 still undefined"]},{"year":2021,"claim":"Two independent studies identified RNF10 as the E3 ligase that monoubiquitylates 40S ribosomal proteins uS3 and uS5 during ribosome-associated quality control, with USP10 as the antagonizing deubiquitylase, establishing the initiation RQC (iRQC) pathway and redefining RNF10's primary molecular function.","evidence":"Genetic screens, quantitative proteomics, PAR-CLIP, ribosome fractionation, knockout/knockdown with ribosome degradation and translation reporter assays","pmids":["34469731","34348161"],"confidence":"High","gaps":["How RNF10 is recruited to stalled ribosomes at a structural level remained unknown","downstream fate of ubiquitinated 40S subunits not fully resolved"]},{"year":2021,"claim":"RNF10 knockdown in macrophages enhanced NF-κB and IRF3 signaling, suggesting a negative regulatory role for RNF10 in innate immune activation, though the direct ubiquitin substrates mediating this effect were not identified.","evidence":"siRNA knockdown in macrophages, pathway reporter assays, cytokine quantification, bacterial clearance assay","pmids":["33249776"],"confidence":"Medium","gaps":["Ubiquitin substrates mediating immune suppression not identified","not independently replicated","unclear if this reflects ribosome QC effects on cytokine translation or a direct signaling role"]},{"year":2024,"claim":"RNF10 ubiquitylation of uS3 was shown to antagonize ribosomal half-mer formation and RNF10 protein levels were found to be dynamically coupled to 40S abundance, revealing a homeostatic feedback mechanism that tunes iRQC to ribosomal subunit stoichiometry.","evidence":"Ribosome fractionation, half-mer analysis, RPS/RPL knockdowns, proteasome inhibition in human cells","pmids":["39609413"],"confidence":"High","gaps":["Proteasomal pathway responsible for RNF10 degradation when 40S levels drop not characterized","structural basis of RNF10's preference for stalled vs. translating 40S not resolved"]},{"year":2024,"claim":"Conservation of RNF10-mediated 40S ubiquitylation was demonstrated in Drosophila, where RNF10 and CNOT4 ubiquitylate eS7/RPS7 upstream of the deubiquitylase OTUD6, extending the pathway to an additional substrate and linking it to alkylation stress responses.","evidence":"Coimmunoprecipitation, OTUD6 catalytic-dead enrichment, genetic epistasis in Drosophila","pmids":["39127721"],"confidence":"Medium","gaps":["Relative contributions of RNF10 vs. CNOT4 on eS7 not separated","mammalian eS7 ubiquitylation by RNF10 not confirmed"]},{"year":2025,"claim":"The downstream effector RIOK3 was identified as the factor that recognizes RNF10-ubiquitylated 40S subunits via a ubiquitin-interacting motif (cryo-EM resolved) and executes 3′-to-5′ 18S rRNA decay, completing the mechanistic pathway from RNF10-mediated ubiquitylation to selective 40S destruction within a GCN2-dependent integrated stress response.","evidence":"Genome-wide CRISPR epistasis screens, selective ribosome profiling, cryo-EM of RIOK3–ubiquitylated 40S complex, 18S rRNA decay assays, amino acid starvation","pmids":["39947182","39947183","40022732"],"confidence":"High","gaps":["How GCN2 activation feeds back to increase RNF10 recruitment to ribosomes not structurally resolved","nuclease(s) executing 18S rRNA degradation downstream of RIOK3 not identified"]},{"year":2025,"claim":"LTN1, the E3 ligase of the canonical 60S-associated RQC pathway, was found to suppress RNF10 expression in a RING-domain-dependent manner, revealing regulatory crosstalk between the two branches of ribosome quality control.","evidence":"Knockout mouse and human cell lines, western blotting, RING domain mutant analysis","pmids":["41451945"],"confidence":"Medium","gaps":["Whether LTN1 directly ubiquitylates RNF10 or acts indirectly not determined","physiological contexts where LTN1–RNF10 crosstalk is rate-limiting not defined"]},{"year":2025,"claim":"Nuclear RNF10 was shown to transcriptionally repress Rbpjk in vascular smooth muscle cells through a non-ubiquitin-ligase mechanism, protecting against vascular calcification in chronic kidney disease, expanding RNF10's transcriptional regulatory repertoire to a new tissue and disease context.","evidence":"Rnf10 knock-in rats, RNA-seq, ChIP-seq, ChIP-qPCR, luciferase reporters, Rbpjk epistasis experiments, proteasome inhibition","pmids":["41988714"],"confidence":"High","gaps":["DNA-binding domain or cofactor enabling RNF10 transcriptional repression of Rbpjk not identified","relationship between RNF10's ribosome QC role and its nuclear transcriptional function in VSMCs not addressed"]},{"year":null,"claim":"A unifying model explaining how RNF10 partitions between its cytoplasmic ribosome quality control function and its nuclear transcriptional roles, and how these dual activities are coordinated in the same cell, remains unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of RNF10 on the ribosome exists","mechanism of RNF10 DNA binding (direct vs. cofactor-dependent) is unknown","tissue-specific regulation of RNF10's ligase vs. transcriptional functions not systematically characterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,2,3,4,5,13]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[9,10,11,15]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[1]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[9]}],"localization":[{"term_id":"GO:0005840","term_label":"ribosome","supporting_discovery_ids":[0,1,2,3,4,5]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[7,8,9,15]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[7]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,2,3,4,5,6]},{"term_id":"R-HSA-8953854","term_label":"Metabolism of RNA","supporting_discovery_ids":[2,3]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[2,4]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[9,10,15]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[7,8]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[9,11]}],"complexes":[],"partners":["RPS3","RPS2","USP10","RIOK3","GRIN2A","MEOX2","LTN1"],"other_free_text":[]},"mechanistic_narrative":"RNF10 is a RING-domain E3 ubiquitin ligase that functions as a central effector of ribosome quality control and additionally serves as a transcriptional regulator in neurons and other cell types. In the initiation ribosome quality control (iRQC) pathway, RNF10 monoubiquitylates 40S ribosomal proteins uS3/RPS3 and uS5/RPS2 on stalled or mRNA-engaged ribosomes, promoting 40S subunit dissociation, half-mer resolution, and selective 40S degradation via RIOK3-mediated 18S nonfunctional rRNA decay within a GCN2-dependent integrated stress response; USP10 acts as the counteracting deubiquitylase, and RNF10 protein levels are dynamically coupled to 40S subunit abundance [PMID:34469731, PMID:34348161, PMID:39947182, PMID:39947183, PMID:39609413]. In neurons, RNF10 resides at excitatory synapses in complex with the NMDAR GluN2A subunit and, upon synaptic activation, undergoes PKC-dependent Ser31 phosphorylation, importin-mediated nuclear translocation, and regulation of transcriptional programs required for LTP maintenance and dendritic spine remodeling [PMID:26977767, PMID:31069631]. Independent of its ligase activity, RNF10 functions as a transcriptional activator of MAG in Schwann cells to promote myelination, interacts with MEOX2 to co-activate p21-dependent cell cycle exit during neuronal differentiation, and represses Rbpjk transcription in vascular smooth muscle cells to inhibit osteogenic transdifferentiation [PMID:18941509, PMID:16335786, PMID:23526782, PMID:41988714]."},"prefetch_data":{"uniprot":{"accession":"Q8N5U6","full_name":"E3 ubiquitin-protein ligase RNF10","aliases":["RING finger protein 10"],"length_aa":811,"mass_kda":89.9,"function":"E3 ubiquitin-protein ligase that catalyzes monoubiquitination of 40S ribosomal proteins RPS2/us5 and RPS3/us3 in response to ribosome stalling (PubMed:34348161, PubMed:34469731, PubMed:39609413, PubMed:39947182, PubMed:39947183, PubMed:40022732). Part of a ribosome quality control that takes place when ribosomes have stalled during translation initiation (iRQC) or elongation (PubMed:34348161, PubMed:34469731, PubMed:39609413, PubMed:39947182, PubMed:39947183, PubMed:40022732). The ribosome quality control is activated in response to ribosome subunit imbalance, amino acid starvation or downstream the EIF2AK4/GCN2-mediated integrated stress response (ISR) (PubMed:39609413, PubMed:39947182, PubMed:39947183, PubMed:40022732). RNF10 acts by mediating monoubiquitination of RPS2/us5 and RPS3/us3: monoubiquitinated RPS2/us5 and RPS3/us3 are then recognized by RIOK3 kinase, leading to 18S non-functional rRNA decay and degradation of the 40S ribosomal subunit (PubMed:34348161, PubMed:34469731, PubMed:39609413, PubMed:39947182, PubMed:39947183, PubMed:40022732). The action of RNF10 in iRQC is counteracted by USP10 (PubMed:34469731)","subcellular_location":"Cytoplasm; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q8N5U6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF10","classification":"Not Classified","n_dependent_lines":4,"n_total_lines":1208,"dependency_fraction":0.0033112582781456954},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"DDOST","stoichiometry":0.2},{"gene":"OST4","stoichiometry":0.2},{"gene":"STT3B","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RNF10","total_profiled":1310},"omim":[{"mim_id":"616388","title":"UBIQUITIN DOMAIN-CONTAINING PROTEIN 1; UBTD1","url":"https://www.omim.org/entry/616388"},{"mim_id":"615998","title":"RING FINGER PROTEIN 10; RNF10","url":"https://www.omim.org/entry/615998"},{"mim_id":"600535","title":"MESENCHYME HOMEOBOX 2; MEOX2","url":"https://www.omim.org/entry/600535"},{"mim_id":"159460","title":"MYELIN-ASSOCIATED GLYCOPROTEIN; MAG","url":"https://www.omim.org/entry/159460"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Mitochondria","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RNF10"},"hgnc":{"alias_symbol":["KIAA0262","RIE2"],"prev_symbol":[]},"alphafold":{"accession":"Q8N5U6","domains":[{"cath_id":"3.30.40.10","chopping":"177-291_316-386","consensus_level":"medium","plddt":91.3092,"start":177,"end":386},{"cath_id":"-","chopping":"505-625","consensus_level":"medium","plddt":94.1603,"start":505,"end":625}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N5U6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N5U6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N5U6-F1-predicted_aligned_error_v6.png","plddt_mean":64.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF10","jax_strain_url":"https://www.jax.org/strain/search?query=RNF10"},"sequence":{"accession":"Q8N5U6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N5U6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N5U6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N5U6"}},"corpus_meta":[{"pmid":"34469731","id":"PMC_34469731","title":"iRQC, a surveillance pathway for 40S ribosomal quality control during mRNA translation initiation.","date":"2021","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/34469731","citation_count":55,"is_preprint":false},{"pmid":"32457162","id":"PMC_32457162","title":"Putative regulators for the continuum of erythroid differentiation revealed by single-cell transcriptome of human BM and UCB cells.","date":"2020","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/32457162","citation_count":52,"is_preprint":false},{"pmid":"24105792","id":"PMC_24105792","title":"Protein microarray characterization of the S-nitrosoproteome.","date":"2013","source":"Molecular & cellular proteomics : MCP","url":"https://pubmed.ncbi.nlm.nih.gov/24105792","citation_count":50,"is_preprint":false},{"pmid":"34348161","id":"PMC_34348161","title":"The E3 ubiquitin ligase RNF10 modifies 40S ribosomal subunits of ribosomes compromised in 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USP10 is the counteracting deubiquitylase. Prolonged uS3/uS5 ubiquitylation leads to selective 40S (but not 60S) ribosomal protein degradation independent of canonical autophagy. This pathway, termed initiation RQC (iRQC), is triggered when scanning or elongating ribosomes are blocked from progressing past the start codon.\",\n      \"method\": \"Genetic screens, quantitative proteomics, knockdown/knockout with ribosome degradation readouts, translation reporter assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal identification of ligase/deubiquitylase pair with multiple orthogonal methods; independently replicated in companion paper (PMID:34348161)\",\n      \"pmids\": [\"34469731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF10 monoubiquitinates RPS2/uS5 and RPS3/uS3 on ribosomes stalled in translation; overexpression of RNF10 phenocopies USP10 knockout by increasing 40S subunit degradation. PAR-CLIP showed RNF10 crosslinks to mRNAs, tRNAs, and 18S rRNA, indicating direct recruitment to stalled ribosomes. ZNF598-independent translation initiation and elongation impairment also contributes to RNF10-mediated ubiquitination.\",\n      \"method\": \"Overexpression/knockout cell lines, PAR-CLIP, ribosome fractionation, ubiquitination assays\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct biochemical identification of substrates with PAR-CLIP crosslinking and ribosome fractionation; independently corroborated by PMID:34469731\",\n      \"pmids\": [\"34348161\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Mammalian 18S nonfunctional rRNA decay (NRD) proceeds through a GCN2-RNF10-RIOK3 axis: nonfunctional 18S rRNA induces translational arrest at start sites, activating GCN2 (integrated stress response), which limits translation initiation; RNF10-mediated ubiquitination of 40S proteins then promotes 40S ribosomal protein turnover and 18S rRNA decay, with RIOK3 binding ubiquitinated 40S subunits to facilitate 18S rRNA degradation.\",\n      \"method\": \"Genome-wide CRISPR genetic interaction screens, selective ribosome profiling, biochemical ISR activation assays, 18S rRNA decay assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — genome-wide epistasis screen plus biochemical validation; published in peer-reviewed journal with multiple orthogonal methods\",\n      \"pmids\": [\"39947182\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RIOK3 specifically recognizes RNF10-ubiquitylated 40S ribosomes through a unique ubiquitin-interacting motif (visualized by cryo-EM), and mediates progressive 3'-to-5' decay of 18S rRNA in the ubiquitylated 40S subunit. Starvation induces selective depletion of 40S ribosomes via RNF10 ubiquitylation followed by RIOK3-dependent degradation.\",\n      \"method\": \"Cryo-EM structure of RIOK3–ubiquitylated 40S complex, genetic knockouts, ribosome degradation assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — cryo-EM structural validation of ubiquitin-interacting motif plus functional genetics; independent parallel study corroborates (PMID:39947182)\",\n      \"pmids\": [\"39947183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Disruption of 60S biogenesis triggers iRQC activation and 40S decay via RNF10-mediated ubiquitylation of uS3/uS5; depletion of the scanning helicase eIF4A1 impairs 40S ubiquitylation, indicating mRNA engagement is required for iRQC. Amino acid starvation also stimulates iRQC-dependent 40S decay. RIOK3 interacts with ubiquitylated 40S subunits to mediate degradation, and both RNF10 and RIOK3 protein levels increase upon iRQC activation (feedforward mechanism).\",\n      \"method\": \"Genetic knockdowns of 60S/40S biogenesis factors, eIF4A1 depletion, amino acid starvation assays, co-immunoprecipitation of RIOK3 with ubiquitylated 40S\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple genetic perturbations with defined phenotypic readouts; corroborated by PMID:39947182 and PMID:39947183\",\n      \"pmids\": [\"40022732\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF10-mediated monoubiquitination of RPS3/uS3 antagonizes ribosomal half-mer formation by promoting dissociation of 40S subunits from ribosomes stalled during both translation elongation and aberrant initiation. RNF10 protein levels are coupled to 40S subunit abundance: knockdown of RPS proteins leads to proteasomal degradation of RNF10, whereas knockdown of RPL proteins causes accumulation of stalled initiating 40S subunits and increased RNF10 levels.\",\n      \"method\": \"Ribosome fractionation, half-mer analysis, RPS/RPL knockdowns, proteasome inhibition, ubiquitination assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal approaches showing mechanistic link between RNF10 activity and ribosomal subunit stoichiometry\",\n      \"pmids\": [\"39609413\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The E3 ubiquitin ligase LTN1 suppresses RNF10 expression in a manner dependent on LTN1's RING domain, revealing crosstalk between RQC-associated E3 ligases as a mechanism coordinating translational surveillance pathways.\",\n      \"method\": \"Knockout mouse and human cell lines, western blotting, RING domain mutant analysis\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic knockouts with domain-level mechanistic dissection; single study\",\n      \"pmids\": [\"41451945\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"RNF10 is a synaptonuclear messenger enriched at excitatory synapses where it associates with the GluN2A subunit of NMDA receptors. Activation of synaptic GluN2A-containing NMDARs and LTP induction cause RNF10 translocation from dendritic segments and spines to the nucleus via importin-dependent long-distance transport. RNF10 silencing prevents LTP maintenance and LTP-dependent structural modifications of dendritic spines.\",\n      \"method\": \"Co-immunoprecipitation with GluN2A, live-cell imaging, importin inhibition, siRNA knockdown with LTP electrophysiology and spine morphology readouts\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, direct imaging of translocation, functional loss-of-function with defined electrophysiological and structural phenotypes\",\n      \"pmids\": [\"26977767\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"PKC-dependent phosphorylation of RNF10 at Ser31 is required for RNF10 detachment from the NMDA receptor GluN2A subunit and subsequent nuclear trafficking. Preventing Ser31 phosphorylation decreases spine density, neuronal branching, and CREB signaling; mimicking stable Ser31 phosphorylation has opposite effects.\",\n      \"method\": \"Phosphomimetic/phosphodead mutants, live-cell imaging, dendritic spine morphometry, CREB reporter assays\",\n      \"journal\": \"Molecular neurobiology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — site-directed mutagenesis of phosphosite with multiple structural and signaling readouts\",\n      \"pmids\": [\"31069631\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"RNF10 binds to a cis-acting element ~160 bp upstream of the MAG transcription start site and acts as a transcriptional activator of the myelin-associated glycoprotein (MAG) gene in Schwann cells. RNF10 knockdown reduces endogenous MAG mRNA and protein; retroviral RNF10 siRNA in Schwann cell–DRG neuron co-cultures inhibits myelin formation.\",\n      \"method\": \"Yeast one-hybrid screen, luciferase reporter assay, siRNA knockdown, retroviral transduction, Schwann cell–DRG neuron myelination co-culture\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — yeast one-hybrid identification of cis-element binding, reporter assays, and functional myelination readout with siRNA; multiple orthogonal methods\",\n      \"pmids\": [\"18941509\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"RNF10 physically interacts with the transcription factor MEOX2 via a central region of MEOX2 (amino acids 101–185); the RING finger domain of RNF10 is not required for MEOX2 binding. RNF10 co-expression enhances MEOX2-mediated activation of the p21WAF1 promoter.\",\n      \"method\": \"Yeast two-hybrid screen, in vitro pull-down, co-immunoprecipitation in mammalian cells, deletion mapping, luciferase reporter assay\",\n      \"journal\": \"Molecular and cellular biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP plus in vitro pull-down and domain mapping with functional reporter assay\",\n      \"pmids\": [\"16335786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RNF10 expression increases upon retinoic acid-induced neuronal differentiation of P19 cells; RNF10 knockdown impairs neuronal differentiation and prevents cell cycle arrest after RA treatment. RNF10 regulates cell cycle exit through upregulation of p21 (but not p27 or p57), and ectopic p21 partially rescues the differentiation defect caused by RNF10 depletion.\",\n      \"method\": \"siRNA knockdown, BrdU incorporation, flow cytometry cell cycle profiling, neuronal marker western blots, p21 rescue experiment\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean knockdown with specific epistasis rescue by p21; single lab study\",\n      \"pmids\": [\"23526782\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RNF10 is identified as a target of S-nitrosylation; eight ubiquitin E3 ligases including RNF10 were found to be potentially S-nitrosylated, suggesting NO-mediated regulation of RNF10's ubiquitin ligase activity.\",\n      \"method\": \"High-density protein microarray with S-nitrosylation-specific labeling and affinity capture; mass spectrometry identification of modified cysteines\",\n      \"journal\": \"Molecular & cellular proteomics\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — proteome-wide screen identifies RNF10 as S-nitrosylated but functional consequence on ligase activity not directly validated\",\n      \"pmids\": [\"24105792\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"In Drosophila, the E3 ligases CNOT4 and RNF10 function upstream of the deubiquitinase OTUD6 to regulate ubiquitination of RPS7/eS7 on the free 40S ribosome, modulating global protein translation and the response to alkylation stress.\",\n      \"method\": \"Coimmunoprecipitation, enrichment of monoubiquitinated proteins from catalytically inactive OTUD6 Drosophila, genetic epistasis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis in Drosophila ortholog context with Co-IP validation; single lab study\",\n      \"pmids\": [\"39127721\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF10 knockdown in macrophages enhances both NF-κB and IRF3 signaling pathways, leading to increased proinflammatory cytokines and type I interferons, and promoting clearance of Listeria monocytogenes, indicating RNF10 acts as a negative regulator of innate immune signaling in macrophages.\",\n      \"method\": \"siRNA knockdown in macrophages, NF-κB/IRF3 pathway reporter assays, cytokine/interferon quantification, bacterial clearance assay\",\n      \"journal\": \"FEBS open bio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — clean knockdown with defined signaling and functional readouts; single lab study\",\n      \"pmids\": [\"33249776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"In the context of vascular calcification in chronic kidney disease, nuclear RNF10 negatively regulates Rbpjk expression in VSMCs through a transcriptional (non-ubiquitin-ligase) mechanism. Proteasome inhibition does not impair RNF10's anticalcific activity. Viral Rbpjk overexpression reverses RNF10's protective effects, while Rbpjk knockdown reduces osteogenic markers, defining an RNF10-Rbpjk regulatory axis.\",\n      \"method\": \"Rnf10 knock-in rats, RNF10 overexpression in VSMCs, RNA-seq, ChIP-seq, ChIP-qPCR, luciferase reporter assays, gain- and loss-of-function for Rbpjk in vivo and in vitro, proteasome inhibition\",\n      \"journal\": \"Arteriosclerosis, thrombosis, and vascular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo knock-in model combined with ChIP-seq, epistasis experiments, and multiple orthogonal validation methods\",\n      \"pmids\": [\"41988714\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"RNF10 interacts with the tumor suppressor RASSF1C in a yeast two-hybrid screen confirmed by in vitro pull-down of bacterially expressed proteins, placing RNF10 within a nuclear interactome network that includes hampin/MSL1 and associated chromatin regulators.\",\n      \"method\": \"Yeast two-hybrid library screen, in vitro pull-down with bacterially expressed proteins\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — yeast two-hybrid plus in vitro pull-down only; functional consequence not established\",\n      \"pmids\": [\"17335777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"RNF10 interacts with HSV-1 tegument protein VP22; co-expression of VP22 with RNF10 relocates RNF10 from its normal subcellular distribution pattern, indicating VP22 modulates RNF10 localization.\",\n      \"method\": \"Yeast two-hybrid, fluorescent protein tagging and co-expression imaging\",\n      \"journal\": \"Archives of virology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — yeast two-hybrid and localization imaging only; no direct functional mechanistic follow-up\",\n      \"pmids\": [\"21424732\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF10 is a RING-domain E3 ubiquitin ligase that functions at multiple levels: in the ribosome quality control pathway it monoubiquitylates 40S small subunit proteins uS3/RPS3 and uS5/RPS2 on stalled or initiating ribosomes (iRQC), promoting 40S subunit dissociation and selective 40S degradation via RIOK3-mediated 18S rRNA decay in a GCN2-dependent integrated stress response; in neurons it serves as a calcium/NMDAR-activated synaptonuclear messenger that undergoes PKC-phosphorylation at Ser31, importin-dependent transport to the nucleus, and regulates transcription required for LTP maintenance and dendritic spine morphology; and in non-neuronal cells it acts as a transcriptional regulator (of MAG in Schwann cells and Rbpjk in VSMCs) and interacts with MEOX2 to activate p21-dependent cell cycle exit.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RNF10 is a RING-domain E3 ubiquitin ligase that functions as a central effector of ribosome quality control and additionally serves as a transcriptional regulator in neurons and other cell types. In the initiation ribosome quality control (iRQC) pathway, RNF10 monoubiquitylates 40S ribosomal proteins uS3/RPS3 and uS5/RPS2 on stalled or mRNA-engaged ribosomes, promoting 40S subunit dissociation, half-mer resolution, and selective 40S degradation via RIOK3-mediated 18S nonfunctional rRNA decay within a GCN2-dependent integrated stress response; USP10 acts as the counteracting deubiquitylase, and RNF10 protein levels are dynamically coupled to 40S subunit abundance [PMID:34469731, PMID:34348161, PMID:39947182, PMID:39947183, PMID:39609413]. In neurons, RNF10 resides at excitatory synapses in complex with the NMDAR GluN2A subunit and, upon synaptic activation, undergoes PKC-dependent Ser31 phosphorylation, importin-mediated nuclear translocation, and regulation of transcriptional programs required for LTP maintenance and dendritic spine remodeling [PMID:26977767, PMID:31069631]. Independent of its ligase activity, RNF10 functions as a transcriptional activator of MAG in Schwann cells to promote myelination, interacts with MEOX2 to co-activate p21-dependent cell cycle exit during neuronal differentiation, and represses Rbpjk transcription in vascular smooth muscle cells to inhibit osteogenic transdifferentiation [PMID:18941509, PMID:16335786, PMID:23526782, PMID:41988714].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Identification of RNF10 as a physical partner of the homeodomain transcription factor MEOX2 established RNF10's first known protein interaction and linked it to p21-dependent transcriptional regulation, raising the question of whether RNF10 functions beyond canonical E3 ligase activity.\",\n      \"evidence\": \"Yeast two-hybrid, co-IP, in vitro pull-down, and p21 promoter reporter assay in mammalian cells\",\n      \"pmids\": [\"16335786\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"RING domain dispensability for MEOX2 binding was shown but the RING domain's catalytic role was not tested in this context\", \"endogenous co-expression and physiological relevance not established\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Discovery that RNF10 directly binds a cis-element upstream of the MAG gene and activates MAG transcription in Schwann cells established RNF10 as a bona fide transcriptional activator controlling peripheral myelination.\",\n      \"evidence\": \"Yeast one-hybrid screen, luciferase reporters, siRNA knockdown, and Schwann cell–DRG neuron myelination co-culture\",\n      \"pmids\": [\"18941509\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which a RING-domain protein binds DNA and activates transcription not resolved\", \"relationship between ubiquitin ligase activity and transcriptional function not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Demonstration that RNF10 is required for retinoic acid-induced neuronal differentiation and cell cycle exit through p21 upregulation connected RNF10's transcriptional co-activator role with MEOX2 to a defined developmental process.\",\n      \"evidence\": \"siRNA knockdown in P19 cells, cell cycle profiling, p21 rescue experiment\",\n      \"pmids\": [\"23526782\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Study performed in a single cell line (P19)\", \"direct transcriptional mechanism on p21 promoter not dissected\", \"whether RNF10 E3 activity is involved in cell cycle regulation unclear\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of RNF10 as an activity-dependent synaptonuclear messenger that translocates from NMDAR/GluN2A complexes to the nucleus upon LTP induction revealed a non-canonical signaling function for an E3 ligase in synaptic plasticity.\",\n      \"evidence\": \"Co-IP with GluN2A, live-cell imaging, importin inhibition, siRNA knockdown with LTP electrophysiology and spine morphology readouts in neurons\",\n      \"pmids\": [\"26977767\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Nuclear transcriptional targets mediating LTP maintenance not identified\", \"whether ubiquitin ligase activity is required for the synaptic function not tested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Pinpointing PKC-dependent phosphorylation of Ser31 as the molecular switch controlling RNF10 detachment from GluN2A and nuclear import resolved the activation mechanism of the synaptonuclear signaling pathway.\",\n      \"evidence\": \"Phosphomimetic/phosphodead mutants, live-cell imaging, spine morphometry, CREB reporter assays in neurons\",\n      \"pmids\": [\"31069631\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Identity of the specific PKC isoform responsible not determined\", \"downstream nuclear gene targets of RNF10 still undefined\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Two independent studies identified RNF10 as the E3 ligase that monoubiquitylates 40S ribosomal proteins uS3 and uS5 during ribosome-associated quality control, with USP10 as the antagonizing deubiquitylase, establishing the initiation RQC (iRQC) pathway and redefining RNF10's primary molecular function.\",\n      \"evidence\": \"Genetic screens, quantitative proteomics, PAR-CLIP, ribosome fractionation, knockout/knockdown with ribosome degradation and translation reporter assays\",\n      \"pmids\": [\"34469731\", \"34348161\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RNF10 is recruited to stalled ribosomes at a structural level remained unknown\", \"downstream fate of ubiquitinated 40S subunits not fully resolved\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"RNF10 knockdown in macrophages enhanced NF-κB and IRF3 signaling, suggesting a negative regulatory role for RNF10 in innate immune activation, though the direct ubiquitin substrates mediating this effect were not identified.\",\n      \"evidence\": \"siRNA knockdown in macrophages, pathway reporter assays, cytokine quantification, bacterial clearance assay\",\n      \"pmids\": [\"33249776\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ubiquitin substrates mediating immune suppression not identified\", \"not independently replicated\", \"unclear if this reflects ribosome QC effects on cytokine translation or a direct signaling role\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"RNF10 ubiquitylation of uS3 was shown to antagonize ribosomal half-mer formation and RNF10 protein levels were found to be dynamically coupled to 40S abundance, revealing a homeostatic feedback mechanism that tunes iRQC to ribosomal subunit stoichiometry.\",\n      \"evidence\": \"Ribosome fractionation, half-mer analysis, RPS/RPL knockdowns, proteasome inhibition in human cells\",\n      \"pmids\": [\"39609413\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Proteasomal pathway responsible for RNF10 degradation when 40S levels drop not characterized\", \"structural basis of RNF10's preference for stalled vs. translating 40S not resolved\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Conservation of RNF10-mediated 40S ubiquitylation was demonstrated in Drosophila, where RNF10 and CNOT4 ubiquitylate eS7/RPS7 upstream of the deubiquitylase OTUD6, extending the pathway to an additional substrate and linking it to alkylation stress responses.\",\n      \"evidence\": \"Coimmunoprecipitation, OTUD6 catalytic-dead enrichment, genetic epistasis in Drosophila\",\n      \"pmids\": [\"39127721\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relative contributions of RNF10 vs. CNOT4 on eS7 not separated\", \"mammalian eS7 ubiquitylation by RNF10 not confirmed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"The downstream effector RIOK3 was identified as the factor that recognizes RNF10-ubiquitylated 40S subunits via a ubiquitin-interacting motif (cryo-EM resolved) and executes 3′-to-5′ 18S rRNA decay, completing the mechanistic pathway from RNF10-mediated ubiquitylation to selective 40S destruction within a GCN2-dependent integrated stress response.\",\n      \"evidence\": \"Genome-wide CRISPR epistasis screens, selective ribosome profiling, cryo-EM of RIOK3–ubiquitylated 40S complex, 18S rRNA decay assays, amino acid starvation\",\n      \"pmids\": [\"39947182\", \"39947183\", \"40022732\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How GCN2 activation feeds back to increase RNF10 recruitment to ribosomes not structurally resolved\", \"nuclease(s) executing 18S rRNA degradation downstream of RIOK3 not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"LTN1, the E3 ligase of the canonical 60S-associated RQC pathway, was found to suppress RNF10 expression in a RING-domain-dependent manner, revealing regulatory crosstalk between the two branches of ribosome quality control.\",\n      \"evidence\": \"Knockout mouse and human cell lines, western blotting, RING domain mutant analysis\",\n      \"pmids\": [\"41451945\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether LTN1 directly ubiquitylates RNF10 or acts indirectly not determined\", \"physiological contexts where LTN1–RNF10 crosstalk is rate-limiting not defined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Nuclear RNF10 was shown to transcriptionally repress Rbpjk in vascular smooth muscle cells through a non-ubiquitin-ligase mechanism, protecting against vascular calcification in chronic kidney disease, expanding RNF10's transcriptional regulatory repertoire to a new tissue and disease context.\",\n      \"evidence\": \"Rnf10 knock-in rats, RNA-seq, ChIP-seq, ChIP-qPCR, luciferase reporters, Rbpjk epistasis experiments, proteasome inhibition\",\n      \"pmids\": [\"41988714\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"DNA-binding domain or cofactor enabling RNF10 transcriptional repression of Rbpjk not identified\", \"relationship between RNF10's ribosome QC role and its nuclear transcriptional function in VSMCs not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A unifying model explaining how RNF10 partitions between its cytoplasmic ribosome quality control function and its nuclear transcriptional roles, and how these dual activities are coordinated in the same cell, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of RNF10 on the ribosome exists\", \"mechanism of RNF10 DNA binding (direct vs. cofactor-dependent) is unknown\", \"tissue-specific regulation of RNF10's ligase vs. transcriptional functions not systematically characterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 5, 13]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [9, 10, 11, 15]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [9]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005840\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 5]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [7, 8, 9, 15]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 5, 6]},\n      {\"term_id\": \"R-HSA-8953854\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [9, 10, 15]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [7, 8]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [9, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"RPS3\",\n      \"RPS2\",\n      \"USP10\",\n      \"RIOK3\",\n      \"GRIN2A\",\n      \"MEOX2\",\n      \"LTN1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}