{"gene":"RELL1","run_date":"2026-06-10T06:43:36","timeline":{"discoveries":[{"year":2005,"finding":"RELL1 and RELL2 are RELT homologues that physically interact with RELT and with each other, as demonstrated by in vitro co-immunoprecipitation. All three proteins co-localize at the plasma membrane. OSR1 (OXSR1) kinase was identified as a RELL1-interacting protein via yeast two-hybrid screen using the intracellular portion of RELL1 as bait, and OSR1 phosphorylates RELL1 (and other RELT family members) in an in vitro kinase assay.","method":"Yeast two-hybrid screen, in vitro co-immunoprecipitation, in vitro kinase assay, co-localization imaging","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (Y2H, Co-IP, in vitro kinase assay, co-localization) in a focused study on RELL1","pmids":["16389068"],"is_preprint":false},{"year":2009,"finding":"Overexpression of RELL1 (and other RELT family members) in HEK 293 epithelial cells induces cell death characterized by cell rounding, lifting, and DNA fragmentation consistent with apoptosis.","method":"Transient transfection overexpression in HEK 293 cells, morphological analysis, DNA fragmentation assay","journal":"Cellular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean overexpression with defined cellular phenotype (apoptosis), two readouts (morphology + DNA fragmentation), single lab","pmids":["19969290"],"is_preprint":false},{"year":2011,"finding":"Phospholipid Scramblase 1 (PLSCR1) physically interacts with RELL1 (and all RELT family members), identified by yeast two-hybrid screen using the intracellular portion of RELL1 as bait, confirmed by co-immunoprecipitation. RELT overexpression alters PLSCR1 intracellular localization. OSR1 phosphorylates PLSCR1 in vitro only in the presence of RELT, suggesting formation of a functional RELT–OSR1–PLSCR1 multiprotein complex.","method":"Yeast two-hybrid screen, co-immunoprecipitation, in vitro kinase assay, co-localization imaging","journal":"Molecular and cellular biochemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal methods (Y2H, Co-IP, in vitro kinase assay, co-localization) demonstrating both interaction and functional consequence","pmids":["22052202"],"is_preprint":false},{"year":2017,"finding":"RELL1 and RELL2 overexpression activates the p38 MAPK pathway more substantially than RELT in HEK-293 cells. This p38 activation by RELL1 is blocked by dominant-negative forms of OSR1 or TRAF2, implicating these molecules downstream of RELL1 signaling.","method":"Transient overexpression in HEK-293 cells, dominant-negative mutant co-transfection, western blotting for p38 activation","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — dominant-negative epistasis plus overexpression phenotype, single lab, two orthogonal approaches","pmids":["28688764"],"is_preprint":false},{"year":2020,"finding":"RELL1 enhances mTOR activity and inhibits autophagy through direct interaction with mTOR in macrophages, promoting Mycobacterium tuberculosis survival. Upregulation of RELL1 increases proinflammatory cytokines (TNF-α and IL-6) but reduces autophagy flux, with net effect of promoting bacterial survival.","method":"RAW264.7 macrophage overexpression, cytokine ELISA, autophagy flux assay, co-immunoprecipitation (direct interaction with mTOR)","journal":"Tuberculosis (Edinburgh, Scotland)","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — functional KO/OE with defined cellular phenotype plus co-IP for mTOR interaction, single lab","pmids":["32090861"],"is_preprint":false},{"year":2020,"finding":"MDFIC (MyoD family inhibitor domain-containing protein), a hematopoietic transcription factor, physically interacts with RELL1 (and other RELT family members), identified by yeast two-hybrid screen using RELL1 as bait. MDFIC co-localizes with RELL1 at the plasma membrane, confirmed by co-immunoprecipitation.","method":"Yeast two-hybrid screen, co-immunoprecipitation with deletion mutants, co-localization imaging","journal":"Biochemistry and biophysics reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Y2H plus Co-IP with deletion mapping, single lab, two orthogonal methods","pmids":["33367115"],"is_preprint":false},{"year":2024,"finding":"Co-transfection of plasmids predicted to block OXSR1 phosphorylation of RELT did not abrogate RELT-induced apoptosis in MDA-MB-231 breast cancer cells, indicating that OSR1/OXSR1-mediated p38 activation is NOT required for RELT family member-induced cell death. Nuclear localization of RELT was also detected in breast cancer cells.","method":"Co-transfection with phosphorylation-blocking mutants, co-immunoprecipitation, immunofluorescence, western blotting, flow cytometry (caspase-3/7 activation, phosphatidylserine externalization)","journal":"Biomedicines","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis/blocking experiment with apoptosis readout plus localization data, single lab, multiple methods","pmids":["39767574"],"is_preprint":false}],"current_model":"RELL1 is a TNF receptor superfamily member (RELT homologue) that localizes to the plasma membrane, physically interacts with RELT, RELL2, OSR1/OXSR1, PLSCR1, and MDFIC via its intracellular domain; is phosphorylated by OSR1 in vitro; activates p38 MAPK signaling through OSR1 and TRAF2; induces apoptosis when overexpressed (by a mechanism that does not require OSR1-mediated phosphorylation); and in macrophages directly interacts with mTOR to suppress autophagy and promote intracellular bacterial survival."},"narrative":{"mechanistic_narrative":"RELL1 is a plasma membrane-associated member of the RELT family of TNF receptor superfamily homologues that functions as a signaling and apoptosis-inducing adaptor [PMID:16389068, PMID:19969290]. Through its intracellular domain it nucleates a multiprotein complex, physically interacting with RELT and RELL2, with the kinase OSR1/OXSR1 (which phosphorylates RELL1 in vitro), with phospholipid scramblase PLSCR1, and with the hematopoietic factor MDFIC, several of which co-localize with RELL1 at the plasma membrane [PMID:16389068, PMID:22052202, PMID:33367115]. Functionally, RELL1 overexpression activates the p38 MAPK pathway through OSR1 and TRAF2 and induces apoptosis, although the cell-death activity does not depend on OSR1/OXSR1-mediated phosphorylation [PMID:28688764, PMID:39767574]. In macrophages, RELL1 directly interacts with mTOR to enhance mTOR activity and suppress autophagy flux, increasing proinflammatory cytokine output and promoting intracellular Mycobacterium tuberculosis survival [PMID:32090861].","teleology":[{"year":2005,"claim":"Established RELL1 as a RELT-family membrane protein and identified its first binding partners and a candidate regulatory kinase, defining a starting framework for its signaling role.","evidence":"Yeast two-hybrid with the RELL1 intracellular domain, in vitro Co-IP, in vitro kinase assay, and co-localization imaging","pmids":["16389068"],"confidence":"High","gaps":["Functional consequence of OSR1 phosphorylation of RELL1 not defined","Interactions shown in vitro/overexpression, not at endogenous levels"]},{"year":2009,"claim":"Showed RELL1 has a cellular output by demonstrating that its overexpression triggers apoptosis, linking the receptor-like protein to programmed cell death.","evidence":"Transient overexpression in HEK 293 cells with morphological scoring and DNA fragmentation assay","pmids":["19969290"],"confidence":"Medium","gaps":["Apoptotic pathway and effector caspases not mapped","Overexpression phenotype not validated by endogenous loss-of-function"]},{"year":2011,"claim":"Extended the interaction network to PLSCR1 and provided evidence for a functional RELT-OSR1-PLSCR1 complex, suggesting RELL1 assembles substrate-presenting modules at the membrane.","evidence":"Yeast two-hybrid, Co-IP, in vitro kinase assay, and co-localization imaging","pmids":["22052202"],"confidence":"High","gaps":["Whether OSR1 phosphorylates PLSCR1 in a RELL1-dependent (versus RELT-dependent) manner not resolved","Downstream consequence of PLSCR1 modification unknown"]},{"year":2017,"claim":"Placed RELL1 upstream of a defined signaling cascade by showing its activation of p38 MAPK requires OSR1 and TRAF2, distinguishing RELL1 signaling potency from RELT.","evidence":"Overexpression with dominant-negative OSR1/TRAF2 epistasis and western blotting for p38 phosphorylation in HEK-293 cells","pmids":["28688764"],"confidence":"Medium","gaps":["Endogenous physiological trigger for p38 activation unknown","Dominant-negative epistasis does not establish direct enzymatic order"]},{"year":2020,"claim":"Revealed an immune/metabolic role distinct from membrane signaling, showing RELL1 directly binds mTOR to suppress autophagy and favor intracellular bacterial persistence in macrophages.","evidence":"RAW264.7 macrophage overexpression, cytokine ELISA, autophagy flux assays, and Co-IP for mTOR interaction","pmids":["32090861"],"confidence":"Medium","gaps":["How a membrane protein engages mTOR mechanistically not defined","Single cell model and single lab"]},{"year":2020,"claim":"Added MDFIC to the RELL1 interactome, expanding partners to a hematopoietic transcription factor at the plasma membrane.","evidence":"Yeast two-hybrid, Co-IP with deletion mapping, and co-localization imaging","pmids":["33367115"],"confidence":"Medium","gaps":["Functional consequence of the RELL1-MDFIC interaction unknown","Interaction not validated at endogenous levels"]},{"year":2024,"claim":"Dissociated RELT-family apoptosis from OSR1/OXSR1-mediated p38 signaling, showing phosphorylation-blocking does not abrogate cell death and revealing a nuclear pool of RELT.","evidence":"Phosphorylation-blocking mutant co-transfection, Co-IP, immunofluorescence, and flow cytometry for caspase-3/7 and phosphatidylserine externalization in MDA-MB-231 cells","pmids":["39767574"],"confidence":"Medium","gaps":["The phosphorylation-independent apoptotic mechanism is not identified","Findings centered on RELT; direct RELL1 generalization not fully tested"]},{"year":null,"claim":"The endogenous physiological ligand/trigger of RELL1 and the molecular basis distinguishing its pro-apoptotic, p38-activating, and mTOR-regulatory functions remain unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No defined extracellular ligand or activation signal","Phosphorylation-independent apoptotic effector unknown","Mechanism linking a membrane protein to mTOR/autophagy not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[0,3]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3,4]}],"localization":[{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[0,2,5]}],"pathway":[{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[3]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[1,6]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[4]}],"complexes":[],"partners":["RELT","RELL2","OXSR1","PLSCR1","MDFIC","TRAF2","MTOR"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8IUW5","full_name":"RELT-like protein 1","aliases":[],"length_aa":271,"mass_kda":29.3,"function":"Induces activation of MAPK14/p38 cascade, when overexpressed (PubMed:28688764). Induces apoptosis, when overexpressed (PubMed:19969290)","subcellular_location":"Cell membrane","url":"https://www.uniprot.org/uniprotkb/Q8IUW5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RELL1","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/RELL1","total_profiled":1310},"omim":[{"mim_id":"611213","title":"RELT-LIKE 2; RELL2","url":"https://www.omim.org/entry/611213"},{"mim_id":"611212","title":"RELT-LIKE 1; RELL1","url":"https://www.omim.org/entry/611212"},{"mim_id":"611211","title":"RECEPTOR EXPRESSED IN LYMPHOID TISSUES; RELT","url":"https://www.omim.org/entry/611211"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"},{"location":"Microtubules","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"brain","ntpm":33.6}],"url":"https://www.proteinatlas.org/search/RELL1"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q8IUW5","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IUW5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IUW5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8IUW5-F1-predicted_aligned_error_v6.png","plddt_mean":60.28},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RELL1","jax_strain_url":"https://www.jax.org/strain/search?query=RELL1"},"sequence":{"accession":"Q8IUW5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8IUW5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8IUW5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8IUW5"}},"corpus_meta":[{"pmid":"22221021","id":"PMC_22221021","title":"A whole-genome association study for pig reproductive traits.","date":"2011","source":"Animal genetics","url":"https://pubmed.ncbi.nlm.nih.gov/22221021","citation_count":131,"is_preprint":false},{"pmid":"32113679","id":"PMC_32113679","title":"Circular RNA circ-RELL1 regulates inflammatory response by miR-6873-3p/MyD88/NF-κB axis in endothelial cells.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32113679","citation_count":52,"is_preprint":false},{"pmid":"35027539","id":"PMC_35027539","title":"Exosomal circRELL1 serves as a miR-637 sponge to modulate gastric cancer progression via regulating autophagy activation.","date":"2022","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/35027539","citation_count":42,"is_preprint":false},{"pmid":"19969290","id":"PMC_19969290","title":"RELT induces cellular death in HEK 293 epithelial cells.","date":"2009","source":"Cellular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/19969290","citation_count":39,"is_preprint":false},{"pmid":"28688764","id":"PMC_28688764","title":"RELT family members activate p38 and induce apoptosis by a mechanism distinct from TNFR1.","date":"2017","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/28688764","citation_count":38,"is_preprint":false},{"pmid":"34646273","id":"PMC_34646273","title":"Surfaceome Proteomic of Glioblastoma Revealed Potential Targets for Immunotherapy.","date":"2021","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/34646273","citation_count":31,"is_preprint":false},{"pmid":"16389068","id":"PMC_16389068","title":"Identification of RELT homologues that associate with RELT and are phosphorylated by OSR1.","date":"2005","source":"Biochemical and biophysical research 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Scotland)","url":"https://pubmed.ncbi.nlm.nih.gov/32090861","citation_count":13,"is_preprint":false},{"pmid":"37893069","id":"PMC_37893069","title":"The RELT Family of Proteins: An Increasing Awareness of Their Importance for Cancer, the Immune System, and Development.","date":"2023","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/37893069","citation_count":12,"is_preprint":false},{"pmid":"32481657","id":"PMC_32481657","title":"A Comparative Quantitative Proteomic Analysis of HCMV-Infected Cells Highlights pUL138 as a Multifunctional Protein.","date":"2020","source":"Molecules (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/32481657","citation_count":11,"is_preprint":false},{"pmid":"37144124","id":"PMC_37144124","title":"Whole-genome selective scans detect genes associated with important phenotypic traits in goat (Capra hircus).","date":"2023","source":"Frontiers in genetics","url":"https://pubmed.ncbi.nlm.nih.gov/37144124","citation_count":10,"is_preprint":false},{"pmid":"33367115","id":"PMC_33367115","title":"RELT stains prominently in B-cell lymphomas and binds the hematopoietic transcription factor MDFIC.","date":"2020","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/33367115","citation_count":6,"is_preprint":false},{"pmid":"39933395","id":"PMC_39933395","title":"Identification of potential drug targets for four site-specific cancers by integrating human plasma proteome with genome.","date":"2025","source":"Journal of pharmaceutical and biomedical analysis","url":"https://pubmed.ncbi.nlm.nih.gov/39933395","citation_count":5,"is_preprint":false},{"pmid":"39130448","id":"PMC_39130448","title":"CircRELL1 promotes osteoarthritis progression by regulating miR-200c-3p.","date":"2024","source":"Heliyon","url":"https://pubmed.ncbi.nlm.nih.gov/39130448","citation_count":2,"is_preprint":false},{"pmid":"39767574","id":"PMC_39767574","title":"RELT Is Upregulated in Breast Cancer and Induces Death in Breast Cancer Cells.","date":"2024","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/39767574","citation_count":1,"is_preprint":false},{"pmid":"39869213","id":"PMC_39869213","title":"The Regulation of Messenger RNAs and Biological Pathways by Long Non-Coding RNAs and Circular RNAs in Ischemic Stroke.","date":"2025","source":"Neurochemical research","url":"https://pubmed.ncbi.nlm.nih.gov/39869213","citation_count":1,"is_preprint":false},{"pmid":"42112773","id":"PMC_42112773","title":"Exploring Immune-Related Gene and Mechanisms in Rosacea Through Transcriptome Analysis and Mendelian Randomization.","date":"2026","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/42112773","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11415,"output_tokens":1925,"usd":0.03156,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":8758,"output_tokens":2519,"usd":0.053382,"stage2_stop_reason":"end_turn"},"total_usd":0.084942,"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\": 2005,\n      \"finding\": \"RELL1 and RELL2 are RELT homologues that physically interact with RELT and with each other, as demonstrated by in vitro co-immunoprecipitation. All three proteins co-localize at the plasma membrane. OSR1 (OXSR1) kinase was identified as a RELL1-interacting protein via yeast two-hybrid screen using the intracellular portion of RELL1 as bait, and OSR1 phosphorylates RELL1 (and other RELT family members) in an in vitro kinase assay.\",\n      \"method\": \"Yeast two-hybrid screen, in vitro co-immunoprecipitation, in vitro kinase assay, co-localization imaging\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (Y2H, Co-IP, in vitro kinase assay, co-localization) in a focused study on RELL1\",\n      \"pmids\": [\"16389068\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Overexpression of RELL1 (and other RELT family members) in HEK 293 epithelial cells induces cell death characterized by cell rounding, lifting, and DNA fragmentation consistent with apoptosis.\",\n      \"method\": \"Transient transfection overexpression in HEK 293 cells, morphological analysis, DNA fragmentation assay\",\n      \"journal\": \"Cellular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean overexpression with defined cellular phenotype (apoptosis), two readouts (morphology + DNA fragmentation), single lab\",\n      \"pmids\": [\"19969290\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Phospholipid Scramblase 1 (PLSCR1) physically interacts with RELL1 (and all RELT family members), identified by yeast two-hybrid screen using the intracellular portion of RELL1 as bait, confirmed by co-immunoprecipitation. RELT overexpression alters PLSCR1 intracellular localization. OSR1 phosphorylates PLSCR1 in vitro only in the presence of RELT, suggesting formation of a functional RELT–OSR1–PLSCR1 multiprotein complex.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation, in vitro kinase assay, co-localization imaging\",\n      \"journal\": \"Molecular and cellular biochemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal methods (Y2H, Co-IP, in vitro kinase assay, co-localization) demonstrating both interaction and functional consequence\",\n      \"pmids\": [\"22052202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RELL1 and RELL2 overexpression activates the p38 MAPK pathway more substantially than RELT in HEK-293 cells. This p38 activation by RELL1 is blocked by dominant-negative forms of OSR1 or TRAF2, implicating these molecules downstream of RELL1 signaling.\",\n      \"method\": \"Transient overexpression in HEK-293 cells, dominant-negative mutant co-transfection, western blotting for p38 activation\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dominant-negative epistasis plus overexpression phenotype, single lab, two orthogonal approaches\",\n      \"pmids\": [\"28688764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RELL1 enhances mTOR activity and inhibits autophagy through direct interaction with mTOR in macrophages, promoting Mycobacterium tuberculosis survival. Upregulation of RELL1 increases proinflammatory cytokines (TNF-α and IL-6) but reduces autophagy flux, with net effect of promoting bacterial survival.\",\n      \"method\": \"RAW264.7 macrophage overexpression, cytokine ELISA, autophagy flux assay, co-immunoprecipitation (direct interaction with mTOR)\",\n      \"journal\": \"Tuberculosis (Edinburgh, Scotland)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — functional KO/OE with defined cellular phenotype plus co-IP for mTOR interaction, single lab\",\n      \"pmids\": [\"32090861\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"MDFIC (MyoD family inhibitor domain-containing protein), a hematopoietic transcription factor, physically interacts with RELL1 (and other RELT family members), identified by yeast two-hybrid screen using RELL1 as bait. MDFIC co-localizes with RELL1 at the plasma membrane, confirmed by co-immunoprecipitation.\",\n      \"method\": \"Yeast two-hybrid screen, co-immunoprecipitation with deletion mutants, co-localization imaging\",\n      \"journal\": \"Biochemistry and biophysics reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Y2H plus Co-IP with deletion mapping, single lab, two orthogonal methods\",\n      \"pmids\": [\"33367115\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Co-transfection of plasmids predicted to block OXSR1 phosphorylation of RELT did not abrogate RELT-induced apoptosis in MDA-MB-231 breast cancer cells, indicating that OSR1/OXSR1-mediated p38 activation is NOT required for RELT family member-induced cell death. Nuclear localization of RELT was also detected in breast cancer cells.\",\n      \"method\": \"Co-transfection with phosphorylation-blocking mutants, co-immunoprecipitation, immunofluorescence, western blotting, flow cytometry (caspase-3/7 activation, phosphatidylserine externalization)\",\n      \"journal\": \"Biomedicines\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis/blocking experiment with apoptosis readout plus localization data, single lab, multiple methods\",\n      \"pmids\": [\"39767574\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RELL1 is a TNF receptor superfamily member (RELT homologue) that localizes to the plasma membrane, physically interacts with RELT, RELL2, OSR1/OXSR1, PLSCR1, and MDFIC via its intracellular domain; is phosphorylated by OSR1 in vitro; activates p38 MAPK signaling through OSR1 and TRAF2; induces apoptosis when overexpressed (by a mechanism that does not require OSR1-mediated phosphorylation); and in macrophages directly interacts with mTOR to suppress autophagy and promote intracellular bacterial survival.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RELL1 is a plasma membrane-associated member of the RELT family of TNF receptor superfamily homologues that functions as a signaling and apoptosis-inducing adaptor [#0, #1]. Through its intracellular domain it nucleates a multiprotein complex, physically interacting with RELT and RELL2, with the kinase OSR1/OXSR1 (which phosphorylates RELL1 in vitro), with phospholipid scramblase PLSCR1, and with the hematopoietic factor MDFIC, several of which co-localize with RELL1 at the plasma membrane [#0, #2, #5]. Functionally, RELL1 overexpression activates the p38 MAPK pathway through OSR1 and TRAF2 and induces apoptosis, although the cell-death activity does not depend on OSR1/OXSR1-mediated phosphorylation [#3, #6]. In macrophages, RELL1 directly interacts with mTOR to enhance mTOR activity and suppress autophagy flux, increasing proinflammatory cytokine output and promoting intracellular Mycobacterium tuberculosis survival [#4].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established RELL1 as a RELT-family membrane protein and identified its first binding partners and a candidate regulatory kinase, defining a starting framework for its signaling role.\",\n      \"evidence\": \"Yeast two-hybrid with the RELL1 intracellular domain, in vitro Co-IP, in vitro kinase assay, and co-localization imaging\",\n      \"pmids\": [\"16389068\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of OSR1 phosphorylation of RELL1 not defined\", \"Interactions shown in vitro/overexpression, not at endogenous levels\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Showed RELL1 has a cellular output by demonstrating that its overexpression triggers apoptosis, linking the receptor-like protein to programmed cell death.\",\n      \"evidence\": \"Transient overexpression in HEK 293 cells with morphological scoring and DNA fragmentation assay\",\n      \"pmids\": [\"19969290\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Apoptotic pathway and effector caspases not mapped\", \"Overexpression phenotype not validated by endogenous loss-of-function\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Extended the interaction network to PLSCR1 and provided evidence for a functional RELT-OSR1-PLSCR1 complex, suggesting RELL1 assembles substrate-presenting modules at the membrane.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, in vitro kinase assay, and co-localization imaging\",\n      \"pmids\": [\"22052202\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether OSR1 phosphorylates PLSCR1 in a RELL1-dependent (versus RELT-dependent) manner not resolved\", \"Downstream consequence of PLSCR1 modification unknown\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Placed RELL1 upstream of a defined signaling cascade by showing its activation of p38 MAPK requires OSR1 and TRAF2, distinguishing RELL1 signaling potency from RELT.\",\n      \"evidence\": \"Overexpression with dominant-negative OSR1/TRAF2 epistasis and western blotting for p38 phosphorylation in HEK-293 cells\",\n      \"pmids\": [\"28688764\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Endogenous physiological trigger for p38 activation unknown\", \"Dominant-negative epistasis does not establish direct enzymatic order\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed an immune/metabolic role distinct from membrane signaling, showing RELL1 directly binds mTOR to suppress autophagy and favor intracellular bacterial persistence in macrophages.\",\n      \"evidence\": \"RAW264.7 macrophage overexpression, cytokine ELISA, autophagy flux assays, and Co-IP for mTOR interaction\",\n      \"pmids\": [\"32090861\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How a membrane protein engages mTOR mechanistically not defined\", \"Single cell model and single lab\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Added MDFIC to the RELL1 interactome, expanding partners to a hematopoietic transcription factor at the plasma membrane.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP with deletion mapping, and co-localization imaging\",\n      \"pmids\": [\"33367115\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the RELL1-MDFIC interaction unknown\", \"Interaction not validated at endogenous levels\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Dissociated RELT-family apoptosis from OSR1/OXSR1-mediated p38 signaling, showing phosphorylation-blocking does not abrogate cell death and revealing a nuclear pool of RELT.\",\n      \"evidence\": \"Phosphorylation-blocking mutant co-transfection, Co-IP, immunofluorescence, and flow cytometry for caspase-3/7 and phosphatidylserine externalization in MDA-MB-231 cells\",\n      \"pmids\": [\"39767574\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The phosphorylation-independent apoptotic mechanism is not identified\", \"Findings centered on RELT; direct RELL1 generalization not fully tested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The endogenous physiological ligand/trigger of RELL1 and the molecular basis distinguishing its pro-apoptotic, p38-activating, and mTOR-regulatory functions remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No defined extracellular ligand or activation signal\", \"Phosphorylation-independent apoptotic effector unknown\", \"Mechanism linking a membrane protein to mTOR/autophagy not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [0, 2, 5]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [1, 6]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RELT\", \"RELL2\", \"OXSR1\", \"PLSCR1\", \"MDFIC\", \"TRAF2\", \"MTOR\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":4,"faith_total":4,"faith_pct":100.0}}