{"gene":"CGRRF1","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":1996,"finding":"CGR19 (CGRRF1) was identified as a p53-induced primary response gene containing a RING-finger domain that is capable of inhibiting the growth of several cell lines when expressed.","method":"Differential RT-PCR in rat embryo fibroblasts with temperature-sensitive p53; growth inhibition assays","journal":"Cancer research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — functional growth-inhibition assay replicated across multiple cell lines, but no molecular mechanism beyond domain annotation; single lab","pmids":["8968090"],"is_preprint":false},{"year":2018,"finding":"CGRRF1 functions as an ER-resident E3 ubiquitin ligase that, together with the E2-conjugating enzyme UBE2J2, ubiquitinates the Wnt cargo receptor Evi (Wls/GPR177) to target it for ERAD via VCP-dependent proteasomal degradation in the absence of Wnt ligands, thereby controlling the scale of Wnt protein secretion.","method":"RNAi knockdown, co-immunoprecipitation, ubiquitination assays, VCP inhibition, epistasis with Porcn, functional Wnt secretion readouts","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in vivo ubiquitination assay, genetic epistasis with UBE2J2/VCP/Porcn, multiple orthogonal methods in one rigorous study","pmids":["29378775"],"is_preprint":false},{"year":2019,"finding":"CGRRF1 acts as an ER-localized RING-domain E3 ubiquitin ligase that directly interacts with EGFR, promotes its K48-linked polyubiquitination, and targets it for proteasomal degradation, thereby suppressing AKT phosphorylation downstream of EGF stimulation and inhibiting breast cancer cell growth in vitro and in vivo.","method":"Co-immunoprecipitation, in vivo ubiquitination assay (K48-linkage specificity), subcellular fractionation, MTT growth assay, xenograft experiment, RING-domain mutant loss-of-function, RPPA substrate identification","journal":"Breast cancer research : BCR","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, linkage-specific ubiquitination assay, RING mutant, xenograft, and RPPA; multiple orthogonal methods in one study","pmids":["31801577"],"is_preprint":false},{"year":2014,"finding":"Overexpression of CGRRF1 inhibits endometrial cancer cell proliferation in vitro, and CGRRF1 expression is induced by metformin in obese rat endometrium, placing it downstream of metformin-responsive signaling.","method":"CGRRF1 overexpression followed by MTT proliferation assay and Western blot; RT-qPCR of rat endometrium after metformin treatment","journal":"Gynecologic oncology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, overexpression proliferation assay without molecular mechanism or pathway placement","pmids":["24680596"],"is_preprint":false},{"year":2024,"finding":"CGRRF1 functions as a ubiquitin E3 ligase for KRAS, mediating K48-linked ubiquitination and proteasomal degradation of KRAS; its suppression by the MSI2-AGO2/miR-30a-3p axis upregulates the KRAS/ERK pathway to promote chemoresistance.","method":"MSI2 RNA-binding assays, AGO2 co-immunoprecipitation, miRNA reporter/loading assays, CGRRF1 ubiquitination assays for KRAS, KRAS protein stability experiments, ERK phosphorylation readouts, pharmacological inhibition","journal":"Neoplasia (New York, N.Y.)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo ubiquitination assay for KRAS substrate identification plus pathway epistasis, single lab, multiple orthogonal methods","pmids":["39522321"],"is_preprint":false}],"current_model":"CGRRF1 is a p53-induced, ER-resident RING-domain E3 ubiquitin ligase that promotes K48-linked ubiquitination and proteasomal degradation of multiple substrates—including the Wnt cargo receptor Evi/Wls (via UBE2J2/VCP-dependent ERAD to regulate Wnt secretion), EGFR (suppressing AKT signaling and breast cancer cell growth), and KRAS (attenuating the KRAS/ERK pathway)—thereby acting as a broadly acting growth suppressor whose loss drives cancer progression and chemoresistance."},"narrative":{"mechanistic_narrative":"CGRRF1 is a p53-induced, ER-resident RING-domain E3 ubiquitin ligase that acts as a growth suppressor by directing K48-linked ubiquitination and proteasomal degradation of multiple membrane and signaling substrates [PMID:8968090, PMID:31801577]. It was first identified as a p53 primary response gene whose expression inhibits the growth of several cell lines [PMID:8968090]. In the endoplasmic reticulum, CGRRF1 partners with the E2-conjugating enzyme UBE2J2 to ubiquitinate the Wnt cargo receptor Evi/Wls, routing it through VCP-dependent ERAD to limit Wnt protein secretion in the absence of Wnt ligands [PMID:29378775]. CGRRF1 also directly binds EGFR and promotes its K48-linked polyubiquitination and degradation, dampening EGF-driven AKT phosphorylation and suppressing breast cancer cell growth in vitro and in xenografts in a manner dependent on an intact RING domain [PMID:31801577]. It further targets KRAS for K48-linked ubiquitination and degradation, attenuating KRAS/ERK signaling; loss of CGRRF1 through an MSI2-AGO2/miR-30a-3p axis upregulates this pathway and drives chemoresistance [PMID:39522321]. Across these substrates CGRRF1 emerges as a broadly acting negative regulator of growth-factor and developmental signaling whose loss contributes to cancer progression.","teleology":[{"year":1996,"claim":"Established CGRRF1 as a p53-responsive, RING-domain gene with intrinsic growth-suppressive activity, framing it as a candidate tumor suppressor before any enzymatic role was known.","evidence":"Differential RT-PCR in rat embryo fibroblasts with temperature-sensitive p53 plus growth-inhibition assays across cell lines","pmids":["8968090"],"confidence":"Medium","gaps":["No molecular substrate or catalytic activity demonstrated","RING-domain function inferred only from sequence annotation"]},{"year":2014,"claim":"Linked CGRRF1 expression to metformin-responsive signaling and antiproliferative effects in endometrial cancer, extending its growth-suppressor phenotype to a new tissue context.","evidence":"CGRRF1 overexpression with MTT proliferation assay and RT-qPCR of metformin-treated rat endometrium","pmids":["24680596"],"confidence":"Low","gaps":["Single lab overexpression assay without molecular mechanism","No defined substrate or signaling pathway","Connection to metformin is correlative"]},{"year":2018,"claim":"Defined CGRRF1's molecular activity for the first time, showing it is an ER-resident E3 ligase that, with UBE2J2 and VCP, degrades the Wnt cargo receptor Evi/Wls to set the scale of Wnt secretion.","evidence":"RNAi, reciprocal Co-IP, in vivo ubiquitination assays, VCP inhibition, and genetic epistasis with Porcn and Wnt-secretion readouts","pmids":["29378775"],"confidence":"High","gaps":["Did not address whether CGRRF1 acts on other ERAD substrates","Regulation of CGRRF1 activity by Wnt ligand status not mechanistically resolved"]},{"year":2019,"claim":"Identified EGFR as a direct CGRRF1 substrate and connected its degradation to suppression of AKT signaling, mechanistically explaining the growth-suppressor phenotype in breast cancer.","evidence":"Reciprocal Co-IP, K48-linkage-specific ubiquitination assay, RING-mutant loss of function, subcellular fractionation, MTT, xenografts, and RPPA substrate identification","pmids":["31801577"],"confidence":"High","gaps":["Whether EGFR degradation occurs via ERAD or another route not specified","E2 partner for EGFR ubiquitination not defined"]},{"year":2024,"claim":"Extended CGRRF1's substrate range to KRAS and embedded it in an MSI2-AGO2/miR-30a-3p regulatory axis, explaining how CGRRF1 loss activates KRAS/ERK signaling to drive chemoresistance.","evidence":"MSI2 RNA-binding and AGO2 Co-IP assays, miRNA reporter assays, KRAS ubiquitination and stability experiments, ERK phosphorylation readouts, and pharmacological inhibition","pmids":["39522321"],"confidence":"Medium","gaps":["Single lab; reciprocal validation of the CGRRF1-KRAS interaction limited","E2 enzyme and subcellular site of KRAS ubiquitination not defined"]},{"year":null,"claim":"It remains unresolved how CGRRF1 selects among its diverse substrates and whether a unifying recruitment or regulatory mechanism governs its activity across the ER and cytosolic signaling pools.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structural model of substrate engagement","Substrate-selection determinants unknown","Interplay between p53 induction and substrate degradation not mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[1,2,4]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,4]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[1,2]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1,2,4]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[2,4]}],"complexes":[],"partners":["UBE2J2","VCP","EVI/WLS","EGFR","KRAS"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q99675","full_name":"Cell growth regulator with RING finger domain protein 1","aliases":["Cell growth regulatory gene 19 protein","RING finger protein 197"],"length_aa":332,"mass_kda":38.2,"function":"Able to inhibit growth in several cell lines","subcellular_location":"Nucleus; Endoplasmic reticulum","url":"https://www.uniprot.org/uniprotkb/Q99675/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/CGRRF1","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CGRRF1","total_profiled":1310},"omim":[{"mim_id":"606138","title":"CELL GROWTH REGULATOR WITH RING FINGER DOMAIN 1; CGRRF1","url":"https://www.omim.org/entry/606138"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/CGRRF1"},"hgnc":{"alias_symbol":["CGR19","RNF197"],"prev_symbol":[]},"alphafold":{"accession":"Q99675","domains":[{"cath_id":"-","chopping":"71-225","consensus_level":"high","plddt":93.7095,"start":71,"end":225},{"cath_id":"3.30.40.10","chopping":"228-233_272-321","consensus_level":"medium","plddt":88.0909,"start":228,"end":321}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99675","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q99675-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q99675-F1-predicted_aligned_error_v6.png","plddt_mean":80.19},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CGRRF1","jax_strain_url":"https://www.jax.org/strain/search?query=CGRRF1"},"sequence":{"accession":"Q99675","fasta_url":"https://rest.uniprot.org/uniprotkb/Q99675.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q99675/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q99675"}},"corpus_meta":[{"pmid":"17029216","id":"PMC_17029216","title":"Novel epigenetically deregulated genes in testicular cancer include homeobox genes and SCGB3A1 (HIN-1).","date":"2006","source":"The Journal of pathology","url":"https://pubmed.ncbi.nlm.nih.gov/17029216","citation_count":76,"is_preprint":false},{"pmid":"8968090","id":"PMC_8968090","title":"Induction of cell growth regulatory genes by p53.","date":"1996","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/8968090","citation_count":72,"is_preprint":false},{"pmid":"11352657","id":"PMC_11352657","title":"Molecular cloning and characterization of RNF26 on human chromosome 11q23 region, encoding a novel RING finger protein with leucine zipper.","date":"2001","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/11352657","citation_count":63,"is_preprint":false},{"pmid":"29378775","id":"PMC_29378775","title":"ERAD-dependent control of the Wnt secretory factor Evi.","date":"2018","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/29378775","citation_count":45,"is_preprint":false},{"pmid":"35702180","id":"PMC_35702180","title":"Dysregulation of MiR-144-5p/RNF187 Axis Contributes To the Progression of Colorectal Cancer.","date":"2022","source":"Journal of translational internal medicine","url":"https://pubmed.ncbi.nlm.nih.gov/35702180","citation_count":38,"is_preprint":false},{"pmid":"12070015","id":"PMC_12070015","title":"Novel transcription factors in human CD34 antigen-positive hematopoietic cells.","date":"2002","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/12070015","citation_count":29,"is_preprint":false},{"pmid":"17485552","id":"PMC_17485552","title":"Gene-expression analysis identifies novel RBL2/p130 target genes in endemic Burkitt lymphoma cell lines and primary tumors.","date":"2007","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/17485552","citation_count":27,"is_preprint":false},{"pmid":"31801577","id":"PMC_31801577","title":"CGRRF1, a growth suppressor, regulates EGFR ubiquitination in breast cancer.","date":"2019","source":"Breast cancer research : BCR","url":"https://pubmed.ncbi.nlm.nih.gov/31801577","citation_count":18,"is_preprint":false},{"pmid":"24680596","id":"PMC_24680596","title":"CGRRF1 as a novel biomarker of tissue response to metformin in the context of obesity.","date":"2014","source":"Gynecologic oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24680596","citation_count":11,"is_preprint":false},{"pmid":"37595490","id":"PMC_37595490","title":"Identification of immunological characteristics and cuproptosis-related molecular clusters in Rheumatoid arthritis.","date":"2023","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/37595490","citation_count":11,"is_preprint":false},{"pmid":"36755350","id":"PMC_36755350","title":"Short-term in vivo testing to discriminate genotoxic carcinogens from non-genotoxic carcinogens and non-carcinogens using next-generation RNA sequencing, DNA microarray, and qPCR.","date":"2023","source":"Genes and environment : the official journal of the Japanese Environmental Mutagen Society","url":"https://pubmed.ncbi.nlm.nih.gov/36755350","citation_count":6,"is_preprint":false},{"pmid":"31367973","id":"PMC_31367973","title":"Rare variants and loci for age-related macular degeneration in the Ohio and Indiana Amish.","date":"2019","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/31367973","citation_count":5,"is_preprint":false},{"pmid":"39522321","id":"PMC_39522321","title":"A novel regulatory axis of MSI2-AGO2/miR-30a-3p-CGRRF1 drives cancer chemoresistance by upregulating the KRAS/ERK pathway.","date":"2024","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/39522321","citation_count":2,"is_preprint":false},{"pmid":"39702344","id":"PMC_39702344","title":"Four functional genotoxic marker genes (Bax, Btg2, Ccng1, and Cdkn1a) discriminate genotoxic hepatocarcinogens from non-genotoxic hepatocarcinogens and non-genotoxic non-hepatocarcinogens in rat public toxicogenomics data, Open TG-GATEs.","date":"2024","source":"Genes and environment : the official journal of the Japanese Environmental Mutagen Society","url":"https://pubmed.ncbi.nlm.nih.gov/39702344","citation_count":1,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10221,"output_tokens":1385,"usd":0.025719,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8049,"output_tokens":2257,"usd":0.048335,"stage2_stop_reason":"end_turn"},"total_usd":0.074054,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1996,\n      \"finding\": \"CGR19 (CGRRF1) was identified as a p53-induced primary response gene containing a RING-finger domain that is capable of inhibiting the growth of several cell lines when expressed.\",\n      \"method\": \"Differential RT-PCR in rat embryo fibroblasts with temperature-sensitive p53; growth inhibition assays\",\n      \"journal\": \"Cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — functional growth-inhibition assay replicated across multiple cell lines, but no molecular mechanism beyond domain annotation; single lab\",\n      \"pmids\": [\"8968090\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CGRRF1 functions as an ER-resident E3 ubiquitin ligase that, together with the E2-conjugating enzyme UBE2J2, ubiquitinates the Wnt cargo receptor Evi (Wls/GPR177) to target it for ERAD via VCP-dependent proteasomal degradation in the absence of Wnt ligands, thereby controlling the scale of Wnt protein secretion.\",\n      \"method\": \"RNAi knockdown, co-immunoprecipitation, ubiquitination assays, VCP inhibition, epistasis with Porcn, functional Wnt secretion readouts\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in vivo ubiquitination assay, genetic epistasis with UBE2J2/VCP/Porcn, multiple orthogonal methods in one rigorous study\",\n      \"pmids\": [\"29378775\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CGRRF1 acts as an ER-localized RING-domain E3 ubiquitin ligase that directly interacts with EGFR, promotes its K48-linked polyubiquitination, and targets it for proteasomal degradation, thereby suppressing AKT phosphorylation downstream of EGF stimulation and inhibiting breast cancer cell growth in vitro and in vivo.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay (K48-linkage specificity), subcellular fractionation, MTT growth assay, xenograft experiment, RING-domain mutant loss-of-function, RPPA substrate identification\",\n      \"journal\": \"Breast cancer research : BCR\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, linkage-specific ubiquitination assay, RING mutant, xenograft, and RPPA; multiple orthogonal methods in one study\",\n      \"pmids\": [\"31801577\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Overexpression of CGRRF1 inhibits endometrial cancer cell proliferation in vitro, and CGRRF1 expression is induced by metformin in obese rat endometrium, placing it downstream of metformin-responsive signaling.\",\n      \"method\": \"CGRRF1 overexpression followed by MTT proliferation assay and Western blot; RT-qPCR of rat endometrium after metformin treatment\",\n      \"journal\": \"Gynecologic oncology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, overexpression proliferation assay without molecular mechanism or pathway placement\",\n      \"pmids\": [\"24680596\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CGRRF1 functions as a ubiquitin E3 ligase for KRAS, mediating K48-linked ubiquitination and proteasomal degradation of KRAS; its suppression by the MSI2-AGO2/miR-30a-3p axis upregulates the KRAS/ERK pathway to promote chemoresistance.\",\n      \"method\": \"MSI2 RNA-binding assays, AGO2 co-immunoprecipitation, miRNA reporter/loading assays, CGRRF1 ubiquitination assays for KRAS, KRAS protein stability experiments, ERK phosphorylation readouts, pharmacological inhibition\",\n      \"journal\": \"Neoplasia (New York, N.Y.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo ubiquitination assay for KRAS substrate identification plus pathway epistasis, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"39522321\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CGRRF1 is a p53-induced, ER-resident RING-domain E3 ubiquitin ligase that promotes K48-linked ubiquitination and proteasomal degradation of multiple substrates—including the Wnt cargo receptor Evi/Wls (via UBE2J2/VCP-dependent ERAD to regulate Wnt secretion), EGFR (suppressing AKT signaling and breast cancer cell growth), and KRAS (attenuating the KRAS/ERK pathway)—thereby acting as a broadly acting growth suppressor whose loss drives cancer progression and chemoresistance.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CGRRF1 is a p53-induced, ER-resident RING-domain E3 ubiquitin ligase that acts as a growth suppressor by directing K48-linked ubiquitination and proteasomal degradation of multiple membrane and signaling substrates [#0, #2]. It was first identified as a p53 primary response gene whose expression inhibits the growth of several cell lines [#0]. In the endoplasmic reticulum, CGRRF1 partners with the E2-conjugating enzyme UBE2J2 to ubiquitinate the Wnt cargo receptor Evi/Wls, routing it through VCP-dependent ERAD to limit Wnt protein secretion in the absence of Wnt ligands [#1]. CGRRF1 also directly binds EGFR and promotes its K48-linked polyubiquitination and degradation, dampening EGF-driven AKT phosphorylation and suppressing breast cancer cell growth in vitro and in xenografts in a manner dependent on an intact RING domain [#2]. It further targets KRAS for K48-linked ubiquitination and degradation, attenuating KRAS/ERK signaling; loss of CGRRF1 through an MSI2-AGO2/miR-30a-3p axis upregulates this pathway and drives chemoresistance [#4]. Across these substrates CGRRF1 emerges as a broadly acting negative regulator of growth-factor and developmental signaling whose loss contributes to cancer progression.\",\n  \"teleology\": [\n    {\n      \"year\": 1996,\n      \"claim\": \"Established CGRRF1 as a p53-responsive, RING-domain gene with intrinsic growth-suppressive activity, framing it as a candidate tumor suppressor before any enzymatic role was known.\",\n      \"evidence\": \"Differential RT-PCR in rat embryo fibroblasts with temperature-sensitive p53 plus growth-inhibition assays across cell lines\",\n      \"pmids\": [\"8968090\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No molecular substrate or catalytic activity demonstrated\", \"RING-domain function inferred only from sequence annotation\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Linked CGRRF1 expression to metformin-responsive signaling and antiproliferative effects in endometrial cancer, extending its growth-suppressor phenotype to a new tissue context.\",\n      \"evidence\": \"CGRRF1 overexpression with MTT proliferation assay and RT-qPCR of metformin-treated rat endometrium\",\n      \"pmids\": [\"24680596\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single lab overexpression assay without molecular mechanism\", \"No defined substrate or signaling pathway\", \"Connection to metformin is correlative\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Defined CGRRF1's molecular activity for the first time, showing it is an ER-resident E3 ligase that, with UBE2J2 and VCP, degrades the Wnt cargo receptor Evi/Wls to set the scale of Wnt secretion.\",\n      \"evidence\": \"RNAi, reciprocal Co-IP, in vivo ubiquitination assays, VCP inhibition, and genetic epistasis with Porcn and Wnt-secretion readouts\",\n      \"pmids\": [\"29378775\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not address whether CGRRF1 acts on other ERAD substrates\", \"Regulation of CGRRF1 activity by Wnt ligand status not mechanistically resolved\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified EGFR as a direct CGRRF1 substrate and connected its degradation to suppression of AKT signaling, mechanistically explaining the growth-suppressor phenotype in breast cancer.\",\n      \"evidence\": \"Reciprocal Co-IP, K48-linkage-specific ubiquitination assay, RING-mutant loss of function, subcellular fractionation, MTT, xenografts, and RPPA substrate identification\",\n      \"pmids\": [\"31801577\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether EGFR degradation occurs via ERAD or another route not specified\", \"E2 partner for EGFR ubiquitination not defined\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended CGRRF1's substrate range to KRAS and embedded it in an MSI2-AGO2/miR-30a-3p regulatory axis, explaining how CGRRF1 loss activates KRAS/ERK signaling to drive chemoresistance.\",\n      \"evidence\": \"MSI2 RNA-binding and AGO2 Co-IP assays, miRNA reporter assays, KRAS ubiquitination and stability experiments, ERK phosphorylation readouts, and pharmacological inhibition\",\n      \"pmids\": [\"39522321\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab; reciprocal validation of the CGRRF1-KRAS interaction limited\", \"E2 enzyme and subcellular site of KRAS ubiquitination not defined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how CGRRF1 selects among its diverse substrates and whether a unifying recruitment or regulatory mechanism governs its activity across the ER and cytosolic signaling pools.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structural model of substrate engagement\", \"Substrate-selection determinants unknown\", \"Interplay between p53 induction and substrate degradation not mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [1, 2, 4]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [1, 2]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1, 2, 4]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [2, 4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"UBE2J2\", \"VCP\", \"Evi/WLS\", \"EGFR\", \"KRAS\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"faith_supported":5,"faith_total":5,"faith_pct":100.0}}