{"gene":"RNF152","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2015,"finding":"RNF152 is a lysosome-anchored E3 ubiquitin ligase that targets RagA for K63-linked polyubiquitination in an amino-acid-sensitive manner, thereby recruiting the GATOR1 GAP complex to RagA and negatively regulating mTORC1 activation at the lysosome. RNF152 knockout results in hyperactivation of mTORC1 and protects cells from amino-acid-starvation-induced autophagy.","method":"Co-immunoprecipitation, ubiquitination assays, RagA ubiquitination-site mutagenesis, RNF152 knockout cells with mTORC1 activity and autophagy readouts","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, site-directed mutagenesis of ubiquitination sites, KO phenotype with multiple orthogonal readouts, replicated in a subsequent study (PMID:38706841)","pmids":["25936802"],"is_preprint":false},{"year":2010,"finding":"RNF152 is a canonical RING finger E3 ubiquitin ligase that localizes to lysosomes (co-localized with LAMP3), undergoes K48-linked polyubiquitination in a manner dependent on its RING finger and transmembrane domains, and promotes apoptosis when overexpressed in HeLa cells.","method":"Co-localization with lysosomal marker LAMP3, in vivo ubiquitination assays with domain deletion/mutation analysis, flow cytometry for apoptosis upon overexpression","journal":"Protein & cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization with functional consequence (apoptosis), domain-deletion analysis of ubiquitination, single lab","pmids":["21203937"],"is_preprint":false},{"year":2020,"finding":"RNF152 positively regulates TLR/IL-1R signaling by directly interacting with the adaptor protein MyD88 and enhancing MyD88 oligomerization, which is required for recruitment of downstream signaling components; this effect is independent of RNF152's E3 ligase activity. RNF152-deficient mice produce less inflammatory cytokines in response to LPS and are more resistant to LPS-induced lethal endotoxemia.","method":"Co-immunoprecipitation of RNF152 with MyD88, MyD88 oligomerization assays, RNF152 overexpression/knockdown with NF-κB reporter assays, RNF152-deficient mouse model with cytokine measurements and endotoxemia model","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP, oligomerization assays, KO mouse model with in vivo phenotype, ubiquitination-independent mechanism confirmed by ligase-dead mutant, multiple orthogonal methods","pmids":["31930677"],"is_preprint":false},{"year":2022,"finding":"RNF152 acts as a negative regulator of Wnt/β-catenin signaling in Xenopus embryos by inhibiting Dishevelled polymerization in an E3 ligase-independent manner (requires the transmembrane domain but not ligase activity). Knockdown of RNF152 enhances Wnt-dependent transcriptional responses and neural crest formation, while overexpression inhibits β-catenin stabilization and target gene expression.","method":"Overexpression and morpholino knockdown in Xenopus embryos, Wnt-responsive reporter assays, ligase-dead and transmembrane-domain truncation mutants, Dishevelled polymerization assays","journal":"BMB reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gain- and loss-of-function in Xenopus with defined molecular mechanism (Dvl polymerization), mutant structure-function analysis, single lab","pmids":["35410636"],"is_preprint":false},{"year":2023,"finding":"RNF152 binds IRAK1 and promotes its K48-linked ubiquitination and proteasomal degradation, thereby reducing IRAK1-dependent transcription of AKR1B10 and suppressing fatty acid oxidation and metastasis in lung adenocarcinoma cells.","method":"Co-immunoprecipitation of RNF152 with IRAK1, ubiquitination assays, protein stability assays, RNF152 overexpression/knockdown with AKR1B10 expression and fatty acid oxidation readouts, IRAK1 rescue experiments, mouse xenograft models","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, rescue with IRAK1 overexpression, in vivo xenograft, single lab","pmids":["37717980"],"is_preprint":false},{"year":2024,"finding":"Fasting-induced RNF152 ubiquitinates the Ragulator subunit p18, leading to its proteasomal degradation; loss of RNF152 increases lysosomal localization of p18 and mTORC1 activity, promoting glycolysis and gemcitabine resistance in gallbladder cancer cells.","method":"Co-immunoprecipitation, ubiquitination assays, proteasome inhibitor experiments, RNF152 overexpression/silencing with mTORC1 activity, glycolysis, and drug sensitivity readouts, lentiviral transfections","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, pharmacological and genetic manipulations with functional readouts, single lab","pmids":["38706841"],"is_preprint":false},{"year":2025,"finding":"RNF152 directly interacts with HSP27 and ubiquitinates it at Lys114, targeting HSP27 for proteasomal degradation; RNF152-mediated HSP27 degradation activates the PI3K/AKT pathway, driving fibroblast-like synoviocyte proliferation and pro-inflammatory cytokine release in temporomandibular joint osteoarthritis.","method":"Immunoprecipitation-mass spectrometry, co-immunoprecipitation, co-localization imaging, protein docking, site-specific ubiquitination (Lys114) assays, in vivo AAV-siRNF152 rat TMJOA model","journal":"Arthritis research & therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP-MS plus Co-IP plus site-specific ubiquitination plus in vivo model, single lab","pmids":["41413572"],"is_preprint":false},{"year":2017,"finding":"Zebrafish rnf152 (ortholog of human RNF152) is required for Delta-Notch signaling and NeuroD expression during brain development; knockdown abolishes NeuroD expression in retinal layers and reduces deltaD, notch1a, notch3, and Notch target genes her4 and ascl1a in midbrain, hindbrain, and rhombomeres.","method":"Morpholino knockdown in zebrafish embryos, whole-mount in situ hybridization for neuroD, deltaD, notch1a, notch3, her4, ascl1a","journal":"Molecules and cells","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — clean loss-of-function with specific transcriptional phenotype, but pathway placement is correlative (expression of Notch components reduced) without direct molecular mechanism for how RNF152 regulates Notch; single lab","pmids":["29276941"],"is_preprint":false},{"year":2024,"finding":"RNF152, as a short-lived lysosomal membrane protein, can be co-opted by bifunctional LYMTAC small molecules to tether KRAS G12D to the lysosomal membrane, inducing KRAS relocalization, inhibition of downstream phospho-ERK signaling, and lysosomal degradation of KRAS G12D.","method":"Heterobifunctional small molecule (LYMTAC) treatment, KRAS relocalization imaging, phospho-ERK signaling assays, lysosomal degradation assays in cell lines","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 / Weak — preprint, RNF152 used as a tool/effector rather than being mechanistically characterized; single lab, no mutagenesis of RNF152 itself","pmids":["bio_10.1101_2024.09.08.611923"],"is_preprint":true},{"year":2021,"finding":"Overexpression of RNF152 in RKO colon cancer cells increases apoptosis and sensitizes cells to NO donor-induced apoptosis, accompanied by decreased expression of anti-apoptotic proteins Bcl-XL and Bcl-2.","method":"RNF152 overexpression in RKO cells, flow cytometry for apoptosis, western blot for Bcl-2 and Bcl-XL","journal":"Zhonghua zhong liu za zhi","confidence":"Low","confidence_rationale":"Tier 3 / Weak — overexpression with downstream protein expression readout, no direct substrate ubiquitination demonstrated, single lab, single method set","pmids":["34034471"],"is_preprint":false}],"current_model":"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that negatively regulates mTORC1 signaling by promoting K63-linked ubiquitination of RagA (recruiting the GATOR1 inhibitory complex) and K48-linked ubiquitination of the Ragulator subunit p18 (targeting it for proteasomal degradation); it also ubiquitinates and destabilizes IRAK1 and HSP27 to modulate inflammatory and oncogenic signaling, and positively regulates TLR/IL-1R signaling through an E3 ligase-independent enhancement of MyD88 oligomerization, while additionally suppressing Wnt/β-catenin signaling by inhibiting Dishevelled polymerization in an E3 ligase-independent, transmembrane-domain-dependent manner."},"narrative":{"mechanistic_narrative":"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that functions as a negative regulator of mTORC1 signaling at the lysosomal surface [PMID:25936802, PMID:21203937]. In an amino-acid-sensitive manner it catalyzes K63-linked polyubiquitination of RagA, which recruits the GATOR1 GAP complex and restrains mTORC1 activation; its loss causes mTORC1 hyperactivation and protects cells from starvation-induced autophagy [PMID:25936802]. RNF152 also targets the Ragulator subunit p18 for ubiquitin-dependent proteasomal degradation, and under fasting this further dampens lysosomal mTORC1 activity [PMID:38706841]. Beyond the mTORC1 axis, RNF152 ubiquitinates additional substrates to control inflammatory and oncogenic outputs: it drives K48-linked ubiquitination and degradation of IRAK1, reducing AKR1B10 transcription and suppressing fatty acid oxidation and metastasis [PMID:37717980], and it ubiquitinates HSP27 at Lys114 to promote its degradation, thereby activating PI3K/AKT signaling [PMID:41413572]. Notably, RNF152 also acts through E3 ligase-independent, transmembrane-domain-dependent mechanisms: it enhances MyD88 oligomerization to positively regulate TLR/IL-1R signaling, with RNF152-deficient mice showing reduced inflammatory cytokines and resistance to endotoxemia [PMID:31930677], and it inhibits Dishevelled polymerization to suppress Wnt/β-catenin signaling [PMID:35410636]. Consistent with its membrane localization and ubiquitination machinery, overexpression promotes apoptosis [PMID:21203937].","teleology":[{"year":2010,"claim":"Established RNF152 as a bona fide lysosomal RING E3 ligase, answering whether it is catalytically active and where it acts in the cell.","evidence":"Co-localization with lysosomal marker LAMP3 and in vivo ubiquitination with RING/transmembrane domain deletion mutants, plus apoptosis readout on overexpression in HeLa cells","pmids":["21203937"],"confidence":"Medium","gaps":["No physiological substrate identified at this stage","Apoptosis phenotype derives from overexpression without endogenous loss-of-function","Single lab"]},{"year":2015,"claim":"Identified the first physiological substrate and pathway, showing RNF152 negatively regulates mTORC1 by K63-ubiquitinating RagA to recruit GATOR1.","evidence":"Reciprocal Co-IP, RagA ubiquitination-site mutagenesis, and RNF152 knockout cells with mTORC1 activity and autophagy readouts","pmids":["25936802"],"confidence":"High","gaps":["Upstream signal coupling RNF152 activity to amino-acid status not fully defined","Deubiquitinase counteracting the K63 chain not identified"]},{"year":2017,"claim":"Extended RNF152 function to development, showing its zebrafish ortholog is required for Delta-Notch signaling and NeuroD expression during neurogenesis.","evidence":"Morpholino knockdown in zebrafish with in situ hybridization for Notch pathway components and target genes","pmids":["29276941"],"confidence":"Medium","gaps":["Pathway placement is correlative — no direct molecular substrate linking RNF152 to Notch","Whether the effect requires E3 ligase activity untested"]},{"year":2020,"claim":"Revealed an E3 ligase-independent role, demonstrating RNF152 positively regulates TLR/IL-1R signaling by promoting MyD88 oligomerization.","evidence":"Co-IP, MyD88 oligomerization and NF-κB reporter assays, ligase-dead mutant, and RNF152-deficient mouse endotoxemia model","pmids":["31930677"],"confidence":"High","gaps":["Structural basis of how RNF152 enhances MyD88 oligomerization unknown","Reconciliation of pro-inflammatory MyD88 role with anti-inflammatory IRAK1 degradation not addressed"]},{"year":2022,"claim":"Defined a second non-catalytic, transmembrane-domain-dependent mechanism, showing RNF152 suppresses Wnt/β-catenin signaling by blocking Dishevelled polymerization.","evidence":"Gain- and loss-of-function in Xenopus embryos, Wnt reporter assays, ligase-dead and transmembrane-truncation mutants, and Dishevelled polymerization assays","pmids":["35410636"],"confidence":"Medium","gaps":["Direct physical interaction interface with Dishevelled not mapped","Relevance in mammalian cells not established","Single lab"]},{"year":2023,"claim":"Identified IRAK1 as a degradative substrate, linking RNF152 to suppression of fatty acid oxidation and metastasis in lung adenocarcinoma.","evidence":"Co-IP, ubiquitination and protein stability assays, IRAK1 rescue, and mouse xenograft models with AKR1B10 and fatty acid oxidation readouts","pmids":["37717980"],"confidence":"Medium","gaps":["Ubiquitination site on IRAK1 not mapped","Single lab","Connection to canonical IRAK1 signaling roles not explored"]},{"year":2024,"claim":"Showed RNF152 degrades the Ragulator subunit p18, providing a fasting-responsive second route to restraining lysosomal mTORC1.","evidence":"Co-IP, ubiquitination assays, proteasome inhibitor experiments, and RNF152 manipulation with mTORC1, glycolysis, and gemcitabine sensitivity readouts in gallbladder cancer cells","pmids":["38706841"],"confidence":"Medium","gaps":["Ubiquitin linkage type and p18 acceptor sites not defined","Relationship between RagA and p18 substrate routes not integrated","Single lab"]},{"year":2024,"claim":"Demonstrated RNF152 can be co-opted as a lysosomal tether by LYMTAC molecules to degrade KRAS G12D, exploiting its short-lived lysosomal membrane localization.","evidence":"Heterobifunctional small molecule treatment with KRAS relocalization imaging, phospho-ERK assays, and lysosomal degradation readouts (preprint)","pmids":["bio_10.1101_2024.09.08.611923"],"confidence":"Low","gaps":["Preprint; RNF152 used as an engineered tool, not mechanistically characterized","No mutagenesis of RNF152 itself","Single lab"]},{"year":2025,"claim":"Identified HSP27 as a site-specific substrate, connecting RNF152-mediated HSP27 degradation to PI3K/AKT activation in osteoarthritis.","evidence":"IP-MS, Co-IP, protein docking, Lys114 site-specific ubiquitination assays, and AAV-siRNF152 rat TMJOA model","pmids":["41413572"],"confidence":"Medium","gaps":["Ubiquitin linkage type not specified","Single lab","Tissue-specificity versus other RNF152 functions not addressed"]},{"year":null,"claim":"How RNF152 toggles between its catalytic (substrate ubiquitination) and non-catalytic (MyD88/Dishevelled scaffolding) modes, and what selects among its diverse substrates in different tissues, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model for substrate/mode selection","Regulation of RNF152 activity by upstream signals (amino acids, fasting) mechanistically incomplete","Structural basis of ligase-independent interactions unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,1,4,5,6]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,4,5,6]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[2,3]}],"localization":[{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[0,1,5]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,2,3,6]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,4,5,6]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,4]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[0]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[1]}],"complexes":[],"partners":["RRAGA","LAMTOR1","MYD88","DVL","IRAK1","HSPB1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N8N0","full_name":"E3 ubiquitin-protein ligase RNF152","aliases":["RING finger protein 152","RING-type E3 ubiquitin transferase RNF152"],"length_aa":203,"mass_kda":22.4,"function":"E3 ubiquitin-protein ligase that acts as a negative regulator of mTORC1 signaling by mediating ubiquitination of RagA/RRAGA and RHEB (PubMed:25936802, PubMed:30514904). Catalyzes 'Lys-63'-linked polyubiquitination of RagA/RRAGA in response to amino acid starvation, thereby regulating mTORC1 signaling (PubMed:25936802). Also mediates monoubiquitination of RHEB, promoting its association with the TSC-TBC complex and subsequent inhibition (PubMed:30514904). Also mediates 'Lys-48'-linked polyubiquitination of target proteins and their subsequent targeting to the proteasome for degradation (PubMed:21203937). Induces apoptosis when overexpressed (PubMed:21203937)","subcellular_location":"Lysosome membrane","url":"https://www.uniprot.org/uniprotkb/Q8N8N0/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF152","classification":"Not Classified","n_dependent_lines":2,"n_total_lines":1208,"dependency_fraction":0.0016556291390728477},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF152","total_profiled":1310},"omim":[{"mim_id":"621029","title":"RING FINGER PROTEIN 183; RNF183","url":"https://www.omim.org/entry/621029"},{"mim_id":"621026","title":"RING FINGER PROTEIN 182; RNF182","url":"https://www.omim.org/entry/621026"},{"mim_id":"616512","title":"RING FINGER PROTEIN 152; RNF152","url":"https://www.omim.org/entry/616512"},{"mim_id":"612194","title":"RAS-RELATED GTP-BINDING PROTEIN A; RRAGA","url":"https://www.omim.org/entry/612194"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"kidney","ntpm":52.8}],"url":"https://www.proteinatlas.org/search/RNF152"},"hgnc":{"alias_symbol":["FLJ39176"],"prev_symbol":[]},"alphafold":{"accession":"Q8N8N0","domains":[{"cath_id":"3.30.40.10","chopping":"5-79","consensus_level":"high","plddt":83.73,"start":5,"end":79}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N8N0","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N8N0-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N8N0-F1-predicted_aligned_error_v6.png","plddt_mean":66.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF152","jax_strain_url":"https://www.jax.org/strain/search?query=RNF152"},"sequence":{"accession":"Q8N8N0","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N8N0.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N8N0/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N8N0"}},"corpus_meta":[{"pmid":"25936802","id":"PMC_25936802","title":"The ubiquitination of rag A GTPase by RNF152 negatively regulates mTORC1 activation.","date":"2015","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/25936802","citation_count":106,"is_preprint":false},{"pmid":"32486221","id":"PMC_32486221","title":"The Role of Tissue-Specific Ubiquitin Ligases, RNF183, RNF186, RNF182 and RNF152, in Disease and Biological Function.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32486221","citation_count":49,"is_preprint":false},{"pmid":"21203937","id":"PMC_21203937","title":"RNF152, a novel lysosome localized E3 ligase with pro-apoptotic activities.","date":"2010","source":"Protein & cell","url":"https://pubmed.ncbi.nlm.nih.gov/21203937","citation_count":35,"is_preprint":false},{"pmid":"31930677","id":"PMC_31930677","title":"RNF152 positively regulates TLR/IL-1R signaling by enhancing MyD88 oligomerization.","date":"2020","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/31930677","citation_count":25,"is_preprint":false},{"pmid":"29276941","id":"PMC_29276941","title":"Rnf152 Is Essential for NeuroD Expression and Delta-Notch Signaling in the Zebrafish Embryos.","date":"2017","source":"Molecules and cells","url":"https://pubmed.ncbi.nlm.nih.gov/29276941","citation_count":12,"is_preprint":false},{"pmid":"33791151","id":"PMC_33791151","title":"Probiotic-derived p8 protein induce apoptosis via regulation of RNF152 in colorectal cancer cells.","date":"2021","source":"American journal of cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/33791151","citation_count":10,"is_preprint":false},{"pmid":"37717980","id":"PMC_37717980","title":"RNF152 Suppresses Fatty Acid Oxidation and Metastasis of Lung Adenocarcinoma by Inhibiting IRAK1-Mediated AKR1B10 Expression.","date":"2023","source":"The American journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/37717980","citation_count":6,"is_preprint":false},{"pmid":"35410636","id":"PMC_35410636","title":"RNF152 negatively regulates Wnt/β-catenin signaling in Xenopus embryos.","date":"2022","source":"BMB reports","url":"https://pubmed.ncbi.nlm.nih.gov/35410636","citation_count":4,"is_preprint":false},{"pmid":"38706841","id":"PMC_38706841","title":"Fasting-induced RNF152 resensitizes gallbladder cancer cells to gemcitabine by inhibiting mTORC1-mediated glycolysis.","date":"2024","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/38706841","citation_count":2,"is_preprint":false},{"pmid":"40474218","id":"PMC_40474218","title":"Mast cell tryptase-PAR2 axis promotes ovarian fibrosis through RNF152-mediated stabilization of Bcl-xL.","date":"2025","source":"Journal of ovarian research","url":"https://pubmed.ncbi.nlm.nih.gov/40474218","citation_count":1,"is_preprint":false},{"pmid":"41413572","id":"PMC_41413572","title":"RNF152-mediated, ubiquitin-dependent degradation of HSP27 activates the PI3K/AKT pathway, driving synovial inflammatory cascades in TMJOA.","date":"2025","source":"Arthritis research & therapy","url":"https://pubmed.ncbi.nlm.nih.gov/41413572","citation_count":0,"is_preprint":false},{"pmid":"34034471","id":"PMC_34034471","title":"[Effect of RNF152 on NO induced apoptosis of colon cancer cells].","date":"2021","source":"Zhonghua zhong liu za zhi [Chinese journal of oncology]","url":"https://pubmed.ncbi.nlm.nih.gov/34034471","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.01.24.634514","title":"Cross cell-type systems genetics reveals the influence of eQTL at multiple points in the developmental trajectory of mouse neural progenitor cells","date":"2025-01-25","source":"bioRxiv","url":"https://doi.org/10.1101/2025.01.24.634514","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.09.08.611923","title":"LYMTACs: Chimeric Small Molecules Repurpose Lysosomal Membrane Proteins for Target Protein Relocalization and Degradation","date":"2024-09-10","source":"bioRxiv","url":"https://doi.org/10.1101/2024.09.08.611923","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":9081,"output_tokens":2871,"usd":0.035154,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10098,"output_tokens":3585,"usd":0.070057,"stage2_stop_reason":"end_turn"},"total_usd":0.105211,"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\": 2015,\n      \"finding\": \"RNF152 is a lysosome-anchored E3 ubiquitin ligase that targets RagA for K63-linked polyubiquitination in an amino-acid-sensitive manner, thereby recruiting the GATOR1 GAP complex to RagA and negatively regulating mTORC1 activation at the lysosome. RNF152 knockout results in hyperactivation of mTORC1 and protects cells from amino-acid-starvation-induced autophagy.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, RagA ubiquitination-site mutagenesis, RNF152 knockout cells with mTORC1 activity and autophagy readouts\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, site-directed mutagenesis of ubiquitination sites, KO phenotype with multiple orthogonal readouts, replicated in a subsequent study (PMID:38706841)\",\n      \"pmids\": [\"25936802\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RNF152 is a canonical RING finger E3 ubiquitin ligase that localizes to lysosomes (co-localized with LAMP3), undergoes K48-linked polyubiquitination in a manner dependent on its RING finger and transmembrane domains, and promotes apoptosis when overexpressed in HeLa cells.\",\n      \"method\": \"Co-localization with lysosomal marker LAMP3, in vivo ubiquitination assays with domain deletion/mutation analysis, flow cytometry for apoptosis upon overexpression\",\n      \"journal\": \"Protein & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization with functional consequence (apoptosis), domain-deletion analysis of ubiquitination, single lab\",\n      \"pmids\": [\"21203937\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF152 positively regulates TLR/IL-1R signaling by directly interacting with the adaptor protein MyD88 and enhancing MyD88 oligomerization, which is required for recruitment of downstream signaling components; this effect is independent of RNF152's E3 ligase activity. RNF152-deficient mice produce less inflammatory cytokines in response to LPS and are more resistant to LPS-induced lethal endotoxemia.\",\n      \"method\": \"Co-immunoprecipitation of RNF152 with MyD88, MyD88 oligomerization assays, RNF152 overexpression/knockdown with NF-κB reporter assays, RNF152-deficient mouse model with cytokine measurements and endotoxemia model\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP, oligomerization assays, KO mouse model with in vivo phenotype, ubiquitination-independent mechanism confirmed by ligase-dead mutant, multiple orthogonal methods\",\n      \"pmids\": [\"31930677\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF152 acts as a negative regulator of Wnt/β-catenin signaling in Xenopus embryos by inhibiting Dishevelled polymerization in an E3 ligase-independent manner (requires the transmembrane domain but not ligase activity). Knockdown of RNF152 enhances Wnt-dependent transcriptional responses and neural crest formation, while overexpression inhibits β-catenin stabilization and target gene expression.\",\n      \"method\": \"Overexpression and morpholino knockdown in Xenopus embryos, Wnt-responsive reporter assays, ligase-dead and transmembrane-domain truncation mutants, Dishevelled polymerization assays\",\n      \"journal\": \"BMB reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gain- and loss-of-function in Xenopus with defined molecular mechanism (Dvl polymerization), mutant structure-function analysis, single lab\",\n      \"pmids\": [\"35410636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF152 binds IRAK1 and promotes its K48-linked ubiquitination and proteasomal degradation, thereby reducing IRAK1-dependent transcription of AKR1B10 and suppressing fatty acid oxidation and metastasis in lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation of RNF152 with IRAK1, ubiquitination assays, protein stability assays, RNF152 overexpression/knockdown with AKR1B10 expression and fatty acid oxidation readouts, IRAK1 rescue experiments, mouse xenograft models\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, rescue with IRAK1 overexpression, in vivo xenograft, single lab\",\n      \"pmids\": [\"37717980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Fasting-induced RNF152 ubiquitinates the Ragulator subunit p18, leading to its proteasomal degradation; loss of RNF152 increases lysosomal localization of p18 and mTORC1 activity, promoting glycolysis and gemcitabine resistance in gallbladder cancer cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, proteasome inhibitor experiments, RNF152 overexpression/silencing with mTORC1 activity, glycolysis, and drug sensitivity readouts, lentiviral transfections\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, pharmacological and genetic manipulations with functional readouts, single lab\",\n      \"pmids\": [\"38706841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF152 directly interacts with HSP27 and ubiquitinates it at Lys114, targeting HSP27 for proteasomal degradation; RNF152-mediated HSP27 degradation activates the PI3K/AKT pathway, driving fibroblast-like synoviocyte proliferation and pro-inflammatory cytokine release in temporomandibular joint osteoarthritis.\",\n      \"method\": \"Immunoprecipitation-mass spectrometry, co-immunoprecipitation, co-localization imaging, protein docking, site-specific ubiquitination (Lys114) assays, in vivo AAV-siRNF152 rat TMJOA model\",\n      \"journal\": \"Arthritis research & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP-MS plus Co-IP plus site-specific ubiquitination plus in vivo model, single lab\",\n      \"pmids\": [\"41413572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Zebrafish rnf152 (ortholog of human RNF152) is required for Delta-Notch signaling and NeuroD expression during brain development; knockdown abolishes NeuroD expression in retinal layers and reduces deltaD, notch1a, notch3, and Notch target genes her4 and ascl1a in midbrain, hindbrain, and rhombomeres.\",\n      \"method\": \"Morpholino knockdown in zebrafish embryos, whole-mount in situ hybridization for neuroD, deltaD, notch1a, notch3, her4, ascl1a\",\n      \"journal\": \"Molecules and cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — clean loss-of-function with specific transcriptional phenotype, but pathway placement is correlative (expression of Notch components reduced) without direct molecular mechanism for how RNF152 regulates Notch; single lab\",\n      \"pmids\": [\"29276941\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF152, as a short-lived lysosomal membrane protein, can be co-opted by bifunctional LYMTAC small molecules to tether KRAS G12D to the lysosomal membrane, inducing KRAS relocalization, inhibition of downstream phospho-ERK signaling, and lysosomal degradation of KRAS G12D.\",\n      \"method\": \"Heterobifunctional small molecule (LYMTAC) treatment, KRAS relocalization imaging, phospho-ERK signaling assays, lysosomal degradation assays in cell lines\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — preprint, RNF152 used as a tool/effector rather than being mechanistically characterized; single lab, no mutagenesis of RNF152 itself\",\n      \"pmids\": [\"bio_10.1101_2024.09.08.611923\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Overexpression of RNF152 in RKO colon cancer cells increases apoptosis and sensitizes cells to NO donor-induced apoptosis, accompanied by decreased expression of anti-apoptotic proteins Bcl-XL and Bcl-2.\",\n      \"method\": \"RNF152 overexpression in RKO cells, flow cytometry for apoptosis, western blot for Bcl-2 and Bcl-XL\",\n      \"journal\": \"Zhonghua zhong liu za zhi\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — overexpression with downstream protein expression readout, no direct substrate ubiquitination demonstrated, single lab, single method set\",\n      \"pmids\": [\"34034471\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that negatively regulates mTORC1 signaling by promoting K63-linked ubiquitination of RagA (recruiting the GATOR1 inhibitory complex) and K48-linked ubiquitination of the Ragulator subunit p18 (targeting it for proteasomal degradation); it also ubiquitinates and destabilizes IRAK1 and HSP27 to modulate inflammatory and oncogenic signaling, and positively regulates TLR/IL-1R signaling through an E3 ligase-independent enhancement of MyD88 oligomerization, while additionally suppressing Wnt/β-catenin signaling by inhibiting Dishevelled polymerization in an E3 ligase-independent, transmembrane-domain-dependent manner.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that functions as a negative regulator of mTORC1 signaling at the lysosomal surface [#0, #1]. In an amino-acid-sensitive manner it catalyzes K63-linked polyubiquitination of RagA, which recruits the GATOR1 GAP complex and restrains mTORC1 activation; its loss causes mTORC1 hyperactivation and protects cells from starvation-induced autophagy [#0]. RNF152 also targets the Ragulator subunit p18 for ubiquitin-dependent proteasomal degradation, and under fasting this further dampens lysosomal mTORC1 activity [#5]. Beyond the mTORC1 axis, RNF152 ubiquitinates additional substrates to control inflammatory and oncogenic outputs: it drives K48-linked ubiquitination and degradation of IRAK1, reducing AKR1B10 transcription and suppressing fatty acid oxidation and metastasis [#4], and it ubiquitinates HSP27 at Lys114 to promote its degradation, thereby activating PI3K/AKT signaling [#6]. Notably, RNF152 also acts through E3 ligase-independent, transmembrane-domain-dependent mechanisms: it enhances MyD88 oligomerization to positively regulate TLR/IL-1R signaling, with RNF152-deficient mice showing reduced inflammatory cytokines and resistance to endotoxemia [#2], and it inhibits Dishevelled polymerization to suppress Wnt/\\u03b2-catenin signaling [#3]. Consistent with its membrane localization and ubiquitination machinery, overexpression promotes apoptosis [#1].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established RNF152 as a bona fide lysosomal RING E3 ligase, answering whether it is catalytically active and where it acts in the cell.\",\n      \"evidence\": \"Co-localization with lysosomal marker LAMP3 and in vivo ubiquitination with RING/transmembrane domain deletion mutants, plus apoptosis readout on overexpression in HeLa cells\",\n      \"pmids\": [\"21203937\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No physiological substrate identified at this stage\", \"Apoptosis phenotype derives from overexpression without endogenous loss-of-function\", \"Single lab\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identified the first physiological substrate and pathway, showing RNF152 negatively regulates mTORC1 by K63-ubiquitinating RagA to recruit GATOR1.\",\n      \"evidence\": \"Reciprocal Co-IP, RagA ubiquitination-site mutagenesis, and RNF152 knockout cells with mTORC1 activity and autophagy readouts\",\n      \"pmids\": [\"25936802\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Upstream signal coupling RNF152 activity to amino-acid status not fully defined\", \"Deubiquitinase counteracting the K63 chain not identified\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Extended RNF152 function to development, showing its zebrafish ortholog is required for Delta-Notch signaling and NeuroD expression during neurogenesis.\",\n      \"evidence\": \"Morpholino knockdown in zebrafish with in situ hybridization for Notch pathway components and target genes\",\n      \"pmids\": [\"29276941\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Pathway placement is correlative — no direct molecular substrate linking RNF152 to Notch\", \"Whether the effect requires E3 ligase activity untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed an E3 ligase-independent role, demonstrating RNF152 positively regulates TLR/IL-1R signaling by promoting MyD88 oligomerization.\",\n      \"evidence\": \"Co-IP, MyD88 oligomerization and NF-\\u03baB reporter assays, ligase-dead mutant, and RNF152-deficient mouse endotoxemia model\",\n      \"pmids\": [\"31930677\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Structural basis of how RNF152 enhances MyD88 oligomerization unknown\", \"Reconciliation of pro-inflammatory MyD88 role with anti-inflammatory IRAK1 degradation not addressed\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined a second non-catalytic, transmembrane-domain-dependent mechanism, showing RNF152 suppresses Wnt/\\u03b2-catenin signaling by blocking Dishevelled polymerization.\",\n      \"evidence\": \"Gain- and loss-of-function in Xenopus embryos, Wnt reporter assays, ligase-dead and transmembrane-truncation mutants, and Dishevelled polymerization assays\",\n      \"pmids\": [\"35410636\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct physical interaction interface with Dishevelled not mapped\", \"Relevance in mammalian cells not established\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified IRAK1 as a degradative substrate, linking RNF152 to suppression of fatty acid oxidation and metastasis in lung adenocarcinoma.\",\n      \"evidence\": \"Co-IP, ubiquitination and protein stability assays, IRAK1 rescue, and mouse xenograft models with AKR1B10 and fatty acid oxidation readouts\",\n      \"pmids\": [\"37717980\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Ubiquitination site on IRAK1 not mapped\", \"Single lab\", \"Connection to canonical IRAK1 signaling roles not explored\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Showed RNF152 degrades the Ragulator subunit p18, providing a fasting-responsive second route to restraining lysosomal mTORC1.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, proteasome inhibitor experiments, and RNF152 manipulation with mTORC1, glycolysis, and gemcitabine sensitivity readouts in gallbladder cancer cells\",\n      \"pmids\": [\"38706841\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Ubiquitin linkage type and p18 acceptor sites not defined\", \"Relationship between RagA and p18 substrate routes not integrated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated RNF152 can be co-opted as a lysosomal tether by LYMTAC molecules to degrade KRAS G12D, exploiting its short-lived lysosomal membrane localization.\",\n      \"evidence\": \"Heterobifunctional small molecule treatment with KRAS relocalization imaging, phospho-ERK assays, and lysosomal degradation readouts (preprint)\",\n      \"pmids\": [\"bio_10.1101_2024.09.08.611923\"],\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Preprint; RNF152 used as an engineered tool, not mechanistically characterized\", \"No mutagenesis of RNF152 itself\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified HSP27 as a site-specific substrate, connecting RNF152-mediated HSP27 degradation to PI3K/AKT activation in osteoarthritis.\",\n      \"evidence\": \"IP-MS, Co-IP, protein docking, Lys114 site-specific ubiquitination assays, and AAV-siRNF152 rat TMJOA model\",\n      \"pmids\": [\"41413572\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Ubiquitin linkage type not specified\", \"Single lab\", \"Tissue-specificity versus other RNF152 functions not addressed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF152 toggles between its catalytic (substrate ubiquitination) and non-catalytic (MyD88/Dishevelled scaffolding) modes, and what selects among its diverse substrates in different tissues, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No unifying model for substrate/mode selection\", \"Regulation of RNF152 activity by upstream signals (amino acids, fasting) mechanistically incomplete\", \"Structural basis of ligase-independent interactions unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 1, 4, 5, 6]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 4, 5, 6]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [0, 1, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 2, 3, 6]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 4, 5, 6]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 4]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RRAGA\", \"LAMTOR1\", \"MYD88\", \"DVL\", \"IRAK1\", \"HSPB1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}