{"gene":"RNF152","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2015,"finding":"RNF152 is a lysosome-anchored E3 ubiquitin ligase that ubiquitinates RagA with K63-linked polyubiquitin chains in an amino-acid-sensitive manner, which recruits the GATOR1 GAP complex to inhibit RagA and thereby negatively regulates mTORC1 activation at the lysosome. RNF152 knockout causes hyperactivation of mTORC1 and protects cells from amino-acid-starvation-induced autophagy.","method":"Co-immunoprecipitation, in vivo ubiquitination assays, site-directed mutagenesis of RagA ubiquitination sites, RNF152 knockout cells, mTORC1 activity assays, autophagy assays","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1-2 — multiple orthogonal methods including mutagenesis, KO, and biochemical reconstitution; foundational paper with >100 citations","pmids":["25936802"],"is_preprint":false},{"year":2010,"finding":"RNF152 is a canonical RING finger E3 ubiquitin ligase localized to lysosomes (co-localizing with the lysosome marker LAMP3). It undergoes K48-linked polyubiquitination dependent on its RING finger and transmembrane domains, and its overexpression induces apoptosis in HeLa cells.","method":"In vivo ubiquitination assay, domain mutagenesis, co-localization with LAMP3 by fluorescence microscopy, overexpression apoptosis assay","journal":"Protein & cell","confidence":"Medium","confidence_rationale":"Tier 2-3 — direct localization and E3 activity demonstrated with mutagenesis; single lab, moderate methods","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 its oligomerization, which is required for downstream NF-κB activation. This function is independent of RNF152's E3 ubiquitin 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, overexpression/knockdown NF-κB reporter assays, RNF152-deficient mice, cytokine ELISA, MyD88 oligomerization assay","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, KO mouse model, multiple orthogonal functional assays, ligase-dead mutant clarifying mechanism","pmids":["31930677"],"is_preprint":false},{"year":2024,"finding":"Fasting-induced RNF152 ubiquitinates the Ragulator subunit p18, leading to its proteasomal degradation, reduction of lysosomal p18 localization, suppression of mTORC1 activity, inhibition of glycolysis, and resensitization of gallbladder cancer cells to gemcitabine.","method":"Ubiquitination assays, lentiviral overexpression/silencing, mTORC1 activity assays, glycolysis measurements, xenograft model","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2-3 — mechanistic pathway established with activators/inhibitors and lentiviral constructs; single lab","pmids":["38706841"],"is_preprint":false},{"year":2023,"finding":"RNF152 directly binds IRAK1 and promotes its ubiquitination and proteasomal degradation, thereby reducing IRAK1-mediated AKR1B10 expression, suppressing fatty acid oxidation, and inhibiting metastasis of lung adenocarcinoma cells.","method":"Co-immunoprecipitation, ubiquitination assay, overexpression/knockdown functional assays, xenograft mouse model, fatty acid oxidation assays","journal":"The American journal of pathology","confidence":"Medium","confidence_rationale":"Tier 2-3 — binding and ubiquitination demonstrated with Co-IP and assays; single lab with in vivo validation","pmids":["37717980"],"is_preprint":false},{"year":2022,"finding":"RNF152 negatively regulates Wnt/β-catenin signaling by inhibiting polymerization of Dishevelled in an E3 ligase-independent manner requiring its transmembrane domain. Overexpression inhibits β-catenin stabilization and Wnt target gene expression; knockdown enhances Wnt responses. RNF152 morphants in Xenopus show defects in neural crest formation.","method":"Overexpression and morpholino knockdown in Xenopus embryos, Wnt-responsive reporter assays, E3 ligase-dead and transmembrane-domain truncation mutants, Dishevelled polymerization assay","journal":"BMB reports","confidence":"Medium","confidence_rationale":"Tier 2-3 — domain mutants define mechanism; Xenopus ortholog with consistent function; single lab","pmids":["35410636"],"is_preprint":false},{"year":2025,"finding":"RNF152 ubiquitinates HSP27 at Lys114, targeting it 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, ubiquitination site mutagenesis, in vivo AAV-siRNF152 delivery in rat TMJOA model","journal":"Arthritis research & therapy","confidence":"Medium","confidence_rationale":"Tier 2 — substrate site mapped by MS and mutagenesis, in vivo validation; single lab","pmids":["41413572"],"is_preprint":false},{"year":2021,"finding":"Overexpression of RNF152 in colon cancer cells (RKO) increases apoptosis and sensitizes cells to NO-donor-induced apoptosis, associated with decreased expression of anti-apoptotic proteins Bcl-2 and Bcl-XL.","method":"Flow cytometry apoptosis assay, western blot for Bcl-2/Bcl-XL, overexpression in RKO cells, NO donor treatment","journal":"Zhonghua zhong liu za zhi [Chinese journal of oncology]","confidence":"Low","confidence_rationale":"Tier 3 — overexpression phenotype with mechanistic inference but no direct ubiquitination substrate identified; single lab, single method set","pmids":["34034471"],"is_preprint":false},{"year":2024,"finding":"RNF152, as a short-lived lysosomal membrane protein, can be co-opted by heterobifunctional LYMTAC molecules to tether and deliver membrane-associated target proteins (including KRAS G12D) to the lysosome for degradation, demonstrating that RNF152 can mediate proximity-induced lysosomal targeting of non-native substrates.","method":"LYMTAC chimeric molecule treatment, KRAS relocalization imaging, phospho-ERK signaling assay, lysosomal degradation assay","journal":"bioRxiv","confidence":"Low","confidence_rationale":"Tier 3 — pharmacological tool study; preprint; demonstrates RNF152 as lysosomal membrane anchor but mechanism is chemical biology, not endogenous","pmids":["bio_10.1101_2024.09.08.611923"],"is_preprint":true}],"current_model":"RNF152 is a RING finger E3 ubiquitin ligase anchored to the lysosomal membrane that negatively regulates mTORC1 signaling by catalyzing K63-linked ubiquitination of RagA (recruiting the GATOR1 GAP complex) and by ubiquitinating the Ragulator subunit p18 for proteasomal degradation; it also ubiquitinates IRAK1 and HSP27 to modulate downstream inflammatory and survival signaling, promotes MyD88 oligomerization (ligase-independently) to enhance TLR/IL-1R–NF-κB responses, and inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization in a transmembrane-domain-dependent but ligase-independent manner."},"narrative":{"teleology":[{"year":2010,"claim":"Establishing RNF152 as a bona fide lysosomal RING E3 ligase answered the basic question of where this uncharacterized RING-finger protein resides and whether it possesses intrinsic ubiquitin ligase activity.","evidence":"Fluorescence co-localization with LAMP3 and in vivo ubiquitination assays with RING/TM mutants in HeLa cells","pmids":["21203937"],"confidence":"Medium","gaps":["No endogenous substrate was identified","Apoptosis phenotype upon overexpression lacked mechanistic detail","Self-ubiquitination versus trans-substrate activity not distinguished"]},{"year":2015,"claim":"Identification of RagA as a direct substrate resolved how RNF152 connects to a major signaling pathway: K63-linked ubiquitination of RagA recruits GATOR1 to inactivate mTORC1, establishing RNF152 as a nutrient-sensing brake on mTORC1.","evidence":"Co-IP, in vivo ubiquitination with chain-type specificity, RagA ubiquitination-site mutants, RNF152-KO cells showing mTORC1 hyperactivation and reduced autophagy","pmids":["25936802"],"confidence":"High","gaps":["How amino acid signals regulate RNF152 activity or stability was not defined","In vivo physiological consequences in animal models not tested","Relationship to the parallel TSC2-mediated regulation of mTORC1 was unresolved"]},{"year":2020,"claim":"Discovery that RNF152 promotes MyD88 oligomerization independently of its E3 ligase activity revealed a scaffolding function and expanded its role beyond nutrient sensing into innate immunity.","evidence":"Reciprocal Co-IP, ligase-dead mutant retaining function, RNF152-deficient mice with impaired LPS-induced cytokine production and resistance to endotoxemia","pmids":["31930677"],"confidence":"High","gaps":["Structural basis for RNF152-MyD88 interaction and oligomerization enhancement unknown","Whether lysosomal localization is required for this innate immune function was not addressed","Potential cross-talk between mTORC1 regulation and TLR signaling through RNF152 not explored"]},{"year":2022,"claim":"Demonstrating that RNF152 inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization — again ligase-independently but requiring its transmembrane domain — established a second non-catalytic scaffolding role and suggested the membrane anchor is critical for protein-protein interaction functions.","evidence":"Wnt reporter assays, ligase-dead and TM-truncation mutants, Dishevelled polymerization assay, Xenopus morpholino knockdown with neural crest defects","pmids":["35410636"],"confidence":"Medium","gaps":["Mechanism by which the transmembrane domain blocks Dvl polymerization not resolved at structural level","Relevance in mammalian Wnt-dependent tissues not confirmed","Whether RNF152 directly binds Dvl or acts indirectly was not fully discriminated"]},{"year":2023,"claim":"Identification of IRAK1 as a ubiquitination substrate targeted for proteasomal degradation linked RNF152's ligase activity to suppression of fatty acid oxidation and metastasis in lung adenocarcinoma.","evidence":"Co-IP, ubiquitination assays, overexpression/knockdown with functional readouts, xenograft mouse model","pmids":["37717980"],"confidence":"Medium","gaps":["Ubiquitin chain type on IRAK1 not specified","Whether IRAK1 degradation feeds back to the TLR–NF-κB axis regulated by RNF152 scaffolding was not tested","Single-lab finding without independent replication"]},{"year":2024,"claim":"Demonstrating that RNF152 ubiquitinates the Ragulator subunit p18 for proteasomal degradation under fasting conditions revealed a second node of mTORC1 suppression at the lysosome, reinforcing the nutrient-sensing role.","evidence":"Ubiquitination assays, lentiviral silencing/overexpression, mTORC1 and glycolysis readouts, gallbladder cancer xenograft model","pmids":["38706841"],"confidence":"Medium","gaps":["Relative contributions of RagA ubiquitination versus p18 degradation to mTORC1 suppression not dissected","Chain type on p18 not specified","Single-lab study in one cancer type"]},{"year":2025,"claim":"Mapping HSP27 Lys114 as a ubiquitination site identified a new substrate and connected RNF152 to PI3K/AKT pathway activation in osteoarthritis-associated synoviocytes.","evidence":"IP-mass spectrometry, site-directed mutagenesis, co-localization imaging, in vivo AAV-siRNF152 in rat TMJOA model","pmids":["41413572"],"confidence":"Medium","gaps":["Ubiquitin chain type on HSP27 not determined","How HSP27 degradation activates PI3K/AKT mechanistically is unclear","Single-lab, single disease model"]},{"year":null,"claim":"How RNF152 switches between its catalytic (E3 ligase) and non-catalytic (scaffolding) functions, and whether these are regulated by nutritional or inflammatory signals in an integrated manner, remains unknown.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural information exists for RNF152 or its complexes","Post-translational regulation of RNF152 itself (beyond self-ubiquitination) is largely uncharacterized","Physiological role in whole-organism nutrient homeostasis has not been tested in RNF152-KO mice under metabolic challenge"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,3,4,6]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[2,5]}],"localization":[{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[0,1,3]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,3,5]},{"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-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,3,4,6]}],"complexes":[],"partners":["RRAGA","MYD88","LAMTOR1","IRAK1","HSPB1","DVL1"],"other_free_text":[]},"mechanistic_narrative":"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that functions as a negative regulator of mTORC1 signaling and modulates innate immune and Wnt pathways through both ligase-dependent and ligase-independent mechanisms. Its canonical E3 activity catalyzes K63-linked ubiquitination of RagA to recruit the GATOR1 GAP complex, thereby suppressing mTORC1 activation at the lysosome in response to amino acid availability, and it ubiquitinates the Ragulator subunit p18 for proteasomal degradation as a second means of mTORC1 inhibition [PMID:25936802, PMID:38706841]. Independent of its ligase activity, RNF152 promotes MyD88 oligomerization to enhance TLR/IL-1R–NF-κB signaling and inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization in a transmembrane-domain-dependent manner [PMID:31930677, PMID:35410636]. RNF152 also ubiquitinates IRAK1 and HSP27 for proteasomal degradation, linking it to suppression of lung adenocarcinoma metastasis and regulation of PI3K/AKT-driven inflammatory responses in synoviocytes [PMID:37717980, PMID:41413572]."},"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":104,"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":9,"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":8312,"output_tokens":2423,"usd":0.030641},"stage2":{"model":"claude-opus-4-6","input_tokens":5736,"output_tokens":2441,"usd":0.134557},"total_usd":0.165198,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","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 ubiquitinates RagA with K63-linked polyubiquitin chains in an amino-acid-sensitive manner, which recruits the GATOR1 GAP complex to inhibit RagA and thereby negatively regulates mTORC1 activation at the lysosome. RNF152 knockout causes hyperactivation of mTORC1 and protects cells from amino-acid-starvation-induced autophagy.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assays, site-directed mutagenesis of RagA ubiquitination sites, RNF152 knockout cells, mTORC1 activity assays, autophagy assays\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — multiple orthogonal methods including mutagenesis, KO, and biochemical reconstitution; foundational paper with >100 citations\",\n      \"pmids\": [\"25936802\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RNF152 is a canonical RING finger E3 ubiquitin ligase localized to lysosomes (co-localizing with the lysosome marker LAMP3). It undergoes K48-linked polyubiquitination dependent on its RING finger and transmembrane domains, and its overexpression induces apoptosis in HeLa cells.\",\n      \"method\": \"In vivo ubiquitination assay, domain mutagenesis, co-localization with LAMP3 by fluorescence microscopy, overexpression apoptosis assay\",\n      \"journal\": \"Protein & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — direct localization and E3 activity demonstrated with mutagenesis; single lab, moderate methods\",\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 its oligomerization, which is required for downstream NF-κB activation. This function is independent of RNF152's E3 ubiquitin 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, overexpression/knockdown NF-κB reporter assays, RNF152-deficient mice, cytokine ELISA, MyD88 oligomerization assay\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, KO mouse model, multiple orthogonal functional assays, ligase-dead mutant clarifying mechanism\",\n      \"pmids\": [\"31930677\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Fasting-induced RNF152 ubiquitinates the Ragulator subunit p18, leading to its proteasomal degradation, reduction of lysosomal p18 localization, suppression of mTORC1 activity, inhibition of glycolysis, and resensitization of gallbladder cancer cells to gemcitabine.\",\n      \"method\": \"Ubiquitination assays, lentiviral overexpression/silencing, mTORC1 activity assays, glycolysis measurements, xenograft model\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — mechanistic pathway established with activators/inhibitors and lentiviral constructs; single lab\",\n      \"pmids\": [\"38706841\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF152 directly binds IRAK1 and promotes its ubiquitination and proteasomal degradation, thereby reducing IRAK1-mediated AKR1B10 expression, suppressing fatty acid oxidation, and inhibiting metastasis of lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, overexpression/knockdown functional assays, xenograft mouse model, fatty acid oxidation assays\",\n      \"journal\": \"The American journal of pathology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — binding and ubiquitination demonstrated with Co-IP and assays; single lab with in vivo validation\",\n      \"pmids\": [\"37717980\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF152 negatively regulates Wnt/β-catenin signaling by inhibiting polymerization of Dishevelled in an E3 ligase-independent manner requiring its transmembrane domain. Overexpression inhibits β-catenin stabilization and Wnt target gene expression; knockdown enhances Wnt responses. RNF152 morphants in Xenopus show defects in neural crest formation.\",\n      \"method\": \"Overexpression and morpholino knockdown in Xenopus embryos, Wnt-responsive reporter assays, E3 ligase-dead and transmembrane-domain truncation mutants, Dishevelled polymerization assay\",\n      \"journal\": \"BMB reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — domain mutants define mechanism; Xenopus ortholog with consistent function; single lab\",\n      \"pmids\": [\"35410636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF152 ubiquitinates HSP27 at Lys114, targeting it 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, ubiquitination site mutagenesis, in vivo AAV-siRNF152 delivery in rat TMJOA model\",\n      \"journal\": \"Arthritis research & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — substrate site mapped by MS and mutagenesis, in vivo validation; single lab\",\n      \"pmids\": [\"41413572\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Overexpression of RNF152 in colon cancer cells (RKO) increases apoptosis and sensitizes cells to NO-donor-induced apoptosis, associated with decreased expression of anti-apoptotic proteins Bcl-2 and Bcl-XL.\",\n      \"method\": \"Flow cytometry apoptosis assay, western blot for Bcl-2/Bcl-XL, overexpression in RKO cells, NO donor treatment\",\n      \"journal\": \"Zhonghua zhong liu za zhi [Chinese journal of oncology]\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — overexpression phenotype with mechanistic inference but no direct ubiquitination substrate identified; single lab, single method set\",\n      \"pmids\": [\"34034471\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF152, as a short-lived lysosomal membrane protein, can be co-opted by heterobifunctional LYMTAC molecules to tether and deliver membrane-associated target proteins (including KRAS G12D) to the lysosome for degradation, demonstrating that RNF152 can mediate proximity-induced lysosomal targeting of non-native substrates.\",\n      \"method\": \"LYMTAC chimeric molecule treatment, KRAS relocalization imaging, phospho-ERK signaling assay, lysosomal degradation assay\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — pharmacological tool study; preprint; demonstrates RNF152 as lysosomal membrane anchor but mechanism is chemical biology, not endogenous\",\n      \"pmids\": [\"bio_10.1101_2024.09.08.611923\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"RNF152 is a RING finger E3 ubiquitin ligase anchored to the lysosomal membrane that negatively regulates mTORC1 signaling by catalyzing K63-linked ubiquitination of RagA (recruiting the GATOR1 GAP complex) and by ubiquitinating the Ragulator subunit p18 for proteasomal degradation; it also ubiquitinates IRAK1 and HSP27 to modulate downstream inflammatory and survival signaling, promotes MyD88 oligomerization (ligase-independently) to enhance TLR/IL-1R–NF-κB responses, and inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization in a transmembrane-domain-dependent but ligase-independent manner.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RNF152 is a lysosome-anchored RING finger E3 ubiquitin ligase that functions as a negative regulator of mTORC1 signaling and modulates innate immune and Wnt pathways through both ligase-dependent and ligase-independent mechanisms. Its canonical E3 activity catalyzes K63-linked ubiquitination of RagA to recruit the GATOR1 GAP complex, thereby suppressing mTORC1 activation at the lysosome in response to amino acid availability, and it ubiquitinates the Ragulator subunit p18 for proteasomal degradation as a second means of mTORC1 inhibition [PMID:25936802, PMID:38706841]. Independent of its ligase activity, RNF152 promotes MyD88 oligomerization to enhance TLR/IL-1R–NF-κB signaling and inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization in a transmembrane-domain-dependent manner [PMID:31930677, PMID:35410636]. RNF152 also ubiquitinates IRAK1 and HSP27 for proteasomal degradation, linking it to suppression of lung adenocarcinoma metastasis and regulation of PI3K/AKT-driven inflammatory responses in synoviocytes [PMID:37717980, PMID:41413572].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Establishing RNF152 as a bona fide lysosomal RING E3 ligase answered the basic question of where this uncharacterized RING-finger protein resides and whether it possesses intrinsic ubiquitin ligase activity.\",\n      \"evidence\": \"Fluorescence co-localization with LAMP3 and in vivo ubiquitination assays with RING/TM mutants in HeLa cells\",\n      \"pmids\": [\"21203937\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No endogenous substrate was identified\",\n        \"Apoptosis phenotype upon overexpression lacked mechanistic detail\",\n        \"Self-ubiquitination versus trans-substrate activity not distinguished\"\n      ]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Identification of RagA as a direct substrate resolved how RNF152 connects to a major signaling pathway: K63-linked ubiquitination of RagA recruits GATOR1 to inactivate mTORC1, establishing RNF152 as a nutrient-sensing brake on mTORC1.\",\n      \"evidence\": \"Co-IP, in vivo ubiquitination with chain-type specificity, RagA ubiquitination-site mutants, RNF152-KO cells showing mTORC1 hyperactivation and reduced autophagy\",\n      \"pmids\": [\"25936802\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"How amino acid signals regulate RNF152 activity or stability was not defined\",\n        \"In vivo physiological consequences in animal models not tested\",\n        \"Relationship to the parallel TSC2-mediated regulation of mTORC1 was unresolved\"\n      ]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Discovery that RNF152 promotes MyD88 oligomerization independently of its E3 ligase activity revealed a scaffolding function and expanded its role beyond nutrient sensing into innate immunity.\",\n      \"evidence\": \"Reciprocal Co-IP, ligase-dead mutant retaining function, RNF152-deficient mice with impaired LPS-induced cytokine production and resistance to endotoxemia\",\n      \"pmids\": [\"31930677\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Structural basis for RNF152-MyD88 interaction and oligomerization enhancement unknown\",\n        \"Whether lysosomal localization is required for this innate immune function was not addressed\",\n        \"Potential cross-talk between mTORC1 regulation and TLR signaling through RNF152 not explored\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating that RNF152 inhibits Wnt/β-catenin signaling by blocking Dishevelled polymerization — again ligase-independently but requiring its transmembrane domain — established a second non-catalytic scaffolding role and suggested the membrane anchor is critical for protein-protein interaction functions.\",\n      \"evidence\": \"Wnt reporter assays, ligase-dead and TM-truncation mutants, Dishevelled polymerization assay, Xenopus morpholino knockdown with neural crest defects\",\n      \"pmids\": [\"35410636\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which the transmembrane domain blocks Dvl polymerization not resolved at structural level\",\n        \"Relevance in mammalian Wnt-dependent tissues not confirmed\",\n        \"Whether RNF152 directly binds Dvl or acts indirectly was not fully discriminated\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identification of IRAK1 as a ubiquitination substrate targeted for proteasomal degradation linked RNF152's ligase activity to suppression of fatty acid oxidation and metastasis in lung adenocarcinoma.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, overexpression/knockdown with functional readouts, xenograft mouse model\",\n      \"pmids\": [\"37717980\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Ubiquitin chain type on IRAK1 not specified\",\n        \"Whether IRAK1 degradation feeds back to the TLR–NF-κB axis regulated by RNF152 scaffolding was not tested\",\n        \"Single-lab finding without independent replication\"\n      ]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrating that RNF152 ubiquitinates the Ragulator subunit p18 for proteasomal degradation under fasting conditions revealed a second node of mTORC1 suppression at the lysosome, reinforcing the nutrient-sensing role.\",\n      \"evidence\": \"Ubiquitination assays, lentiviral silencing/overexpression, mTORC1 and glycolysis readouts, gallbladder cancer xenograft model\",\n      \"pmids\": [\"38706841\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Relative contributions of RagA ubiquitination versus p18 degradation to mTORC1 suppression not dissected\",\n        \"Chain type on p18 not specified\",\n        \"Single-lab study in one cancer type\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Mapping HSP27 Lys114 as a ubiquitination site identified a new substrate and connected RNF152 to PI3K/AKT pathway activation in osteoarthritis-associated synoviocytes.\",\n      \"evidence\": \"IP-mass spectrometry, site-directed mutagenesis, co-localization imaging, in vivo AAV-siRNF152 in rat TMJOA model\",\n      \"pmids\": [\"41413572\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Ubiquitin chain type on HSP27 not determined\",\n        \"How HSP27 degradation activates PI3K/AKT mechanistically is unclear\",\n        \"Single-lab, single disease model\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF152 switches between its catalytic (E3 ligase) and non-catalytic (scaffolding) functions, and whether these are regulated by nutritional or inflammatory signals in an integrated manner, remains unknown.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural information exists for RNF152 or its complexes\",\n        \"Post-translational regulation of RNF152 itself (beyond self-ubiquitination) is largely uncharacterized\",\n        \"Physiological role in whole-organism nutrient homeostasis has not been tested in RNF152-KO mice under metabolic challenge\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 3, 4, 6]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [2, 5]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [0, 1, 3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0162582\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 3, 5]},\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-392499\", \"supporting_discovery_ids\": [0, 3, 4, 6]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"RRAGA\",\n      \"MYD88\",\n      \"LAMTOR1\",\n      \"IRAK1\",\n      \"HSPB1\",\n      \"DVL1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}