{"gene":"SHKBP1","run_date":"2026-06-10T07:46:31","timeline":{"discoveries":[{"year":2000,"finding":"SHKBP1 (SB1) was identified as a novel binding partner of SETA (SH3KBP1/CIN85), interacting via SETA's N-terminal SH3 domain. The interaction was confirmed by in vitro confrontation assays and co-immunoprecipitation. SHKBP1 shares 55% amino acid identity with the renal tumor antigen NY-REN-45.","method":"Yeast two-hybrid screening, in vitro confrontation assay, co-immunoprecipitation","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP and in vitro pulldown in a single lab, two orthogonal methods confirming the interaction","pmids":["11152963"],"is_preprint":false},{"year":2019,"finding":"SHKBP1 is a direct target of miR-499a and functions downstream of TGFβ-induced EMT to regulate EGFR activity. Increased SHKBP1 expression co-opts TGFβ-induced AKT activation and contributes to an EGFR-independent, AKT-activated kinase switch that confers resistance to erlotinib in osteosarcoma stem cell-like cells. Luciferase reporter assays confirmed miR-499a directly targets SHKBP1.","method":"Luciferase reporter assay, western blot, in vitro and in vivo erlotinib resistance assays, miRNA overexpression/knockdown","journal":"Journal of experimental & clinical cancer research : CR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — luciferase reporter for direct miR-499a targeting confirmed, functional rescue in vitro and in vivo, single lab with multiple orthogonal methods","pmids":["31138318"],"is_preprint":false},{"year":2021,"finding":"Shkbp1 knockout mice showed significantly reduced lung tumor formation after chemical induction, and Shkbp1 loss in cancer cell lines inhibited migration and invasion while overexpression promoted these phenotypes. SHKBP1 was associated with epithelial-mesenchymal transition (EMT) markers, placing it upstream of EMT-driven tumor progression. SHKBP1 was also found to be expressed in immune cells, and its knockout reduced tumor growth in subcutaneous transplant and metastasis models.","method":"Shkbp1 knockout mouse model, chemical tumor induction, cell migration/invasion assays, overexpression/knockdown cell lines, subcutaneous and tail-vein transplant tumor models","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean genetic KO with defined tumor phenotype, multiple in vitro and in vivo models in a single lab","pmids":["34112919"],"is_preprint":false},{"year":2023,"finding":"Shkbp1 knockout mice exhibited abnormal spleen architecture (disrupted red and white pulp) and increased CD8+ T cell numbers and function in the spleen. Knockout of Shkbp1 enhanced CD8+ T cell infiltration into tumor tissue and inhibited subcutaneous melanoma tumor growth. Adenoviral-mediated Shkbp1 knockout similarly increased CD8+ T cell numbers and suppressed tumor growth, establishing a role for SHKBP1 in regulating CD8+ T cell differentiation and anti-tumor immunity.","method":"Shkbp1 knockout mouse model, subcutaneous melanoma model, adenoviral Shkbp1 targeting, flow cytometry of immune cells, histology","journal":"Molecular immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic KO plus adenoviral KO with orthogonal tumor models, single lab","pmids":["37343421"],"is_preprint":false},{"year":2025,"finding":"SHKBP1 functions as a Cullin-3 E3 ubiquitin ligase adaptor that inhibits SQSTM1/p62 oligomerization through a direct protein-protein interaction mapped outside of p62 bodies. This interaction limits p62 assembly into phase-separated p62 bodies and prevents sequestration and degradation of Keap1, thereby affecting nuclear translocation of Nrf2 and the cellular antioxidant response. This mechanism is independent of SHKBP1's potential role in ubiquitination.","method":"Protein-protein interaction mapping, co-immunoprecipitation, live-cell imaging of p62 bodies, Keap1/Nrf2 reporter assays, domain mapping","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — mechanistic dissection with interaction mapping, functional phase-separation assays, and antioxidant pathway readouts; published in peer-reviewed journal (replicated/extended from preprint)","pmids":["41649860"],"is_preprint":false},{"year":2025,"finding":"SHKBP1 interacts with SQSTM1/p62 outside p62 bodies to inhibit p62 oligomerization and incorporation into p62 bodies, preventing Keap1 sequestration. This is a non-ubiquitination-based function of the Cullin-3 adaptor SHKBP1 in regulating p62 phase separation and oxidative stress responses (preprint version of the above peer-reviewed paper).","method":"Protein-protein interaction mapping, p62 body phase-separation assays, Keap1 sequestration assays","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — preprint with multiple methods; superseded by peer-reviewed version (PMID 41649860)","pmids":["39896619"],"is_preprint":true}],"current_model":"SHKBP1 (SB1) is a Cullin-3 E3 ubiquitin ligase adaptor that binds SETA/CIN85 via its SH3 domain; it inhibits SQSTM1/p62 oligomerization and phase separation through a direct protein-protein interaction, thereby preventing Keap1 sequestration and modulating the Nrf2-mediated antioxidant response independent of its ubiquitination function; it also acts downstream of TGFβ/miR-499a to regulate EGFR activity and promote EMT-associated kinase switching, and plays a role in CD8+ T cell differentiation and anti-tumor immunity."},"narrative":{"mechanistic_narrative":"SHKBP1 (SB1) is a Cullin-3 E3 ubiquitin ligase adaptor that controls the SQSTM1/p62–Keap1–Nrf2 antioxidant axis and contributes to tumor progression and anti-tumor immunity [PMID:41649860, PMID:34112919]. It was first identified as a binding partner of SETA/SH3KBP1/CIN85, engaging the N-terminal SH3 domain of CIN85 [PMID:11152963]. Mechanistically, SHKBP1 binds p62 directly at a site outside p62 bodies and inhibits p62 oligomerization and its incorporation into phase-separated p62 bodies; by limiting p62 assembly it prevents sequestration and degradation of Keap1, thereby restraining Nrf2 nuclear translocation and the cellular antioxidant response, a function that operates independently of SHKBP1's ubiquitination activity [PMID:41649860]. In cancer, SHKBP1 acts downstream of TGFβ-induced EMT as a direct target of miR-499a, co-opting AKT activation to drive an EGFR-independent kinase switch that confers erlotinib resistance [PMID:31138318], and its genetic loss reduces tumor formation, migration, and invasion [PMID:34112919]. SHKBP1 also regulates CD8+ T cell differentiation and anti-tumor immunity, as its knockout increases CD8+ T cell numbers and tumor infiltration while suppressing tumor growth [PMID:37343421].","teleology":[{"year":2000,"claim":"Established the first molecular context for SHKBP1 by identifying it as a physical partner of the CIN85/SETA adaptor, implicating it in adaptor-mediated signaling.","evidence":"Yeast two-hybrid, in vitro confrontation assay, and co-immunoprecipitation mapping the interaction to CIN85's N-terminal SH3 domain","pmids":["11152963"],"confidence":"Medium","gaps":["Functional consequence of the SHKBP1–CIN85 interaction not defined","No cellular pathway or downstream effect demonstrated","Single-lab interaction without orthogonal structural validation"]},{"year":2019,"claim":"Connected SHKBP1 to therapy resistance by showing it is a direct miR-499a target acting downstream of TGFβ-induced EMT to drive AKT-dependent, EGFR-independent kinase switching.","evidence":"Luciferase reporter assays, western blot, and in vitro/in vivo erlotinib resistance assays in osteosarcoma stem cell-like cells","pmids":["31138318"],"confidence":"Medium","gaps":["Molecular mechanism by which SHKBP1 promotes AKT activation not resolved","Direct link to EGFR regulation at biochemical level unclear","Restricted to a single tumor model"]},{"year":2021,"claim":"Provided genetic causal evidence that SHKBP1 promotes tumor initiation and metastatic phenotypes via EMT, moving it from a candidate to a functional driver.","evidence":"Shkbp1 knockout mouse with chemical tumor induction plus migration/invasion and transplant/metastasis cell models","pmids":["34112919"],"confidence":"Medium","gaps":["Molecular effector linking SHKBP1 to EMT markers not identified","Cell-autonomous versus immune-microenvironment contributions not separated"]},{"year":2023,"claim":"Defined a previously unknown immunoregulatory role, showing SHKBP1 restrains CD8+ T cell differentiation and anti-tumor immunity.","evidence":"Germline and adenoviral Shkbp1 knockout with subcutaneous melanoma model, flow cytometry, and spleen histology","pmids":["37343421"],"confidence":"Medium","gaps":["Molecular pathway in T cells controlled by SHKBP1 not identified","Whether the immune phenotype reflects T cell-intrinsic function not resolved"]},{"year":2025,"claim":"Resolved a direct molecular mechanism: SHKBP1 binds p62 to block its oligomerization and phase separation, thereby preventing Keap1 sequestration and tuning Nrf2-dependent antioxidant responses independent of ubiquitination.","evidence":"Protein-protein interaction and domain mapping, live-cell imaging of p62 bodies, and Keap1/Nrf2 reporter assays (peer-reviewed, extended from a 2025 bioRxiv preprint)","pmids":["41649860","39896619"],"confidence":"High","gaps":["Whether the antioxidant function explains the tumor and immune phenotypes not established","Role of SHKBP1's Cullin-3 adaptor/ubiquitination activity in vivo not defined","Structural basis of the SHKBP1–p62 interaction not determined"]},{"year":null,"claim":"How SHKBP1's distinct activities — CIN85 binding, p62/Keap1/Nrf2 regulation, AKT-driven kinase switching, and CD8+ T cell control — are mechanistically unified remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No single mechanism links the antioxidant, EMT, and immune phenotypes","Substrates of the SHKBP1–Cullin-3 complex unidentified","Tissue- and cell-type-specific functions not delineated"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[4]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[4]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[4]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[3]}],"complexes":[],"partners":["SH3KBP1","SQSTM1","KEAP1","CUL3"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8TBC3","full_name":"SH3KBP1-binding protein 1","aliases":["SETA-binding protein 1"],"length_aa":707,"mass_kda":76.3,"function":"Inhibits CBL-SH3KBP1 complex mediated down-regulation of EGFR signaling by sequestration of SH3KBP1. Binds to SH3KBP1 and prevents its interaction with CBL and inhibits translocation of SH3KBP1 to EGFR containing vesicles upon EGF stimulation","subcellular_location":"Lysosome","url":"https://www.uniprot.org/uniprotkb/Q8TBC3/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/SHKBP1","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"SNX27","stoichiometry":10.0},{"gene":"LAMP1","stoichiometry":0.2},{"gene":"MIF","stoichiometry":0.2},{"gene":"VPS35","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/SHKBP1","total_profiled":1310},"omim":[{"mim_id":"617322","title":"SH3KBP1-BINDING PROTEIN 1; SHKBP1","url":"https://www.omim.org/entry/617322"},{"mim_id":"603956","title":"CERVICAL CANCER","url":"https://www.omim.org/entry/603956"},{"mim_id":"300374","title":"SH3-DOMAIN KINASE-BINDING PROTEIN 1; SH3KBP1","url":"https://www.omim.org/entry/300374"},{"mim_id":"131550","title":"EPIDERMAL GROWTH FACTOR RECEPTOR; EGFR","url":"https://www.omim.org/entry/131550"},{"mim_id":"116810","title":"CATHEPSIN B; CTSB","url":"https://www.omim.org/entry/116810"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cell Junctions","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"lymphoid tissue","ntpm":124.1}],"url":"https://www.proteinatlas.org/search/SHKBP1"},"hgnc":{"alias_symbol":["PP203","Sb1"],"prev_symbol":[]},"alphafold":{"accession":"Q8TBC3","domains":[{"cath_id":"3.30.710.10","chopping":"19-116","consensus_level":"high","plddt":89.6336,"start":19,"end":116}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8TBC3","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8TBC3-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8TBC3-F1-predicted_aligned_error_v6.png","plddt_mean":73.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=SHKBP1","jax_strain_url":"https://www.jax.org/strain/search?query=SHKBP1"},"sequence":{"accession":"Q8TBC3","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8TBC3.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8TBC3/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8TBC3"}},"corpus_meta":[{"pmid":"30236955","id":"PMC_30236955","title":"Bacteriophage Sb-1 enhances antibiotic activity against biofilm, degrades exopolysaccharide matrix and targets persisters of Staphylococcus aureus.","date":"2018","source":"International journal of antimicrobial agents","url":"https://pubmed.ncbi.nlm.nih.gov/30236955","citation_count":160,"is_preprint":false},{"pmid":"17046713","id":"PMC_17046713","title":"PGRP-SB1: an N-acetylmuramoyl L-alanine amidase with antibacterial activity.","date":"2006","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17046713","citation_count":120,"is_preprint":false},{"pmid":"29499945","id":"PMC_29499945","title":"circ-SHKBP1 Regulates the Angiogenesis of U87 Glioma-Exposed Endothelial Cells through miR-544a/FOXP1 and miR-379/FOXP2 Pathways.","date":"2017","source":"Molecular therapy. 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The interaction was confirmed by in vitro confrontation assays and co-immunoprecipitation. SHKBP1 shares 55% amino acid identity with the renal tumor antigen NY-REN-45.\",\n      \"method\": \"Yeast two-hybrid screening, in vitro confrontation assay, co-immunoprecipitation\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP and in vitro pulldown in a single lab, two orthogonal methods confirming the interaction\",\n      \"pmids\": [\"11152963\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"SHKBP1 is a direct target of miR-499a and functions downstream of TGFβ-induced EMT to regulate EGFR activity. Increased SHKBP1 expression co-opts TGFβ-induced AKT activation and contributes to an EGFR-independent, AKT-activated kinase switch that confers resistance to erlotinib in osteosarcoma stem cell-like cells. Luciferase reporter assays confirmed miR-499a directly targets SHKBP1.\",\n      \"method\": \"Luciferase reporter assay, western blot, in vitro and in vivo erlotinib resistance assays, miRNA overexpression/knockdown\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — luciferase reporter for direct miR-499a targeting confirmed, functional rescue in vitro and in vivo, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"31138318\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Shkbp1 knockout mice showed significantly reduced lung tumor formation after chemical induction, and Shkbp1 loss in cancer cell lines inhibited migration and invasion while overexpression promoted these phenotypes. SHKBP1 was associated with epithelial-mesenchymal transition (EMT) markers, placing it upstream of EMT-driven tumor progression. SHKBP1 was also found to be expressed in immune cells, and its knockout reduced tumor growth in subcutaneous transplant and metastasis models.\",\n      \"method\": \"Shkbp1 knockout mouse model, chemical tumor induction, cell migration/invasion assays, overexpression/knockdown cell lines, subcutaneous and tail-vein transplant tumor models\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean genetic KO with defined tumor phenotype, multiple in vitro and in vivo models in a single lab\",\n      \"pmids\": [\"34112919\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Shkbp1 knockout mice exhibited abnormal spleen architecture (disrupted red and white pulp) and increased CD8+ T cell numbers and function in the spleen. Knockout of Shkbp1 enhanced CD8+ T cell infiltration into tumor tissue and inhibited subcutaneous melanoma tumor growth. Adenoviral-mediated Shkbp1 knockout similarly increased CD8+ T cell numbers and suppressed tumor growth, establishing a role for SHKBP1 in regulating CD8+ T cell differentiation and anti-tumor immunity.\",\n      \"method\": \"Shkbp1 knockout mouse model, subcutaneous melanoma model, adenoviral Shkbp1 targeting, flow cytometry of immune cells, histology\",\n      \"journal\": \"Molecular immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic KO plus adenoviral KO with orthogonal tumor models, single lab\",\n      \"pmids\": [\"37343421\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SHKBP1 functions as a Cullin-3 E3 ubiquitin ligase adaptor that inhibits SQSTM1/p62 oligomerization through a direct protein-protein interaction mapped outside of p62 bodies. This interaction limits p62 assembly into phase-separated p62 bodies and prevents sequestration and degradation of Keap1, thereby affecting nuclear translocation of Nrf2 and the cellular antioxidant response. This mechanism is independent of SHKBP1's potential role in ubiquitination.\",\n      \"method\": \"Protein-protein interaction mapping, co-immunoprecipitation, live-cell imaging of p62 bodies, Keap1/Nrf2 reporter assays, domain mapping\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — mechanistic dissection with interaction mapping, functional phase-separation assays, and antioxidant pathway readouts; published in peer-reviewed journal (replicated/extended from preprint)\",\n      \"pmids\": [\"41649860\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SHKBP1 interacts with SQSTM1/p62 outside p62 bodies to inhibit p62 oligomerization and incorporation into p62 bodies, preventing Keap1 sequestration. This is a non-ubiquitination-based function of the Cullin-3 adaptor SHKBP1 in regulating p62 phase separation and oxidative stress responses (preprint version of the above peer-reviewed paper).\",\n      \"method\": \"Protein-protein interaction mapping, p62 body phase-separation assays, Keap1 sequestration assays\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — preprint with multiple methods; superseded by peer-reviewed version (PMID 41649860)\",\n      \"pmids\": [\"39896619\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"SHKBP1 (SB1) is a Cullin-3 E3 ubiquitin ligase adaptor that binds SETA/CIN85 via its SH3 domain; it inhibits SQSTM1/p62 oligomerization and phase separation through a direct protein-protein interaction, thereby preventing Keap1 sequestration and modulating the Nrf2-mediated antioxidant response independent of its ubiquitination function; it also acts downstream of TGFβ/miR-499a to regulate EGFR activity and promote EMT-associated kinase switching, and plays a role in CD8+ T cell differentiation and anti-tumor immunity.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"SHKBP1 (SB1) is a Cullin-3 E3 ubiquitin ligase adaptor that controls the SQSTM1/p62–Keap1–Nrf2 antioxidant axis and contributes to tumor progression and anti-tumor immunity [#4, #2]. It was first identified as a binding partner of SETA/SH3KBP1/CIN85, engaging the N-terminal SH3 domain of CIN85 [#0]. Mechanistically, SHKBP1 binds p62 directly at a site outside p62 bodies and inhibits p62 oligomerization and its incorporation into phase-separated p62 bodies; by limiting p62 assembly it prevents sequestration and degradation of Keap1, thereby restraining Nrf2 nuclear translocation and the cellular antioxidant response, a function that operates independently of SHKBP1's ubiquitination activity [#4]. In cancer, SHKBP1 acts downstream of TGFβ-induced EMT as a direct target of miR-499a, co-opting AKT activation to drive an EGFR-independent kinase switch that confers erlotinib resistance [#1], and its genetic loss reduces tumor formation, migration, and invasion [#2]. SHKBP1 also regulates CD8+ T cell differentiation and anti-tumor immunity, as its knockout increases CD8+ T cell numbers and tumor infiltration while suppressing tumor growth [#3].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"Established the first molecular context for SHKBP1 by identifying it as a physical partner of the CIN85/SETA adaptor, implicating it in adaptor-mediated signaling.\",\n      \"evidence\": \"Yeast two-hybrid, in vitro confrontation assay, and co-immunoprecipitation mapping the interaction to CIN85's N-terminal SH3 domain\",\n      \"pmids\": [\"11152963\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional consequence of the SHKBP1–CIN85 interaction not defined\",\n        \"No cellular pathway or downstream effect demonstrated\",\n        \"Single-lab interaction without orthogonal structural validation\"\n      ]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Connected SHKBP1 to therapy resistance by showing it is a direct miR-499a target acting downstream of TGFβ-induced EMT to drive AKT-dependent, EGFR-independent kinase switching.\",\n      \"evidence\": \"Luciferase reporter assays, western blot, and in vitro/in vivo erlotinib resistance assays in osteosarcoma stem cell-like cells\",\n      \"pmids\": [\"31138318\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular mechanism by which SHKBP1 promotes AKT activation not resolved\",\n        \"Direct link to EGFR regulation at biochemical level unclear\",\n        \"Restricted to a single tumor model\"\n      ]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Provided genetic causal evidence that SHKBP1 promotes tumor initiation and metastatic phenotypes via EMT, moving it from a candidate to a functional driver.\",\n      \"evidence\": \"Shkbp1 knockout mouse with chemical tumor induction plus migration/invasion and transplant/metastasis cell models\",\n      \"pmids\": [\"34112919\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular effector linking SHKBP1 to EMT markers not identified\",\n        \"Cell-autonomous versus immune-microenvironment contributions not separated\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined a previously unknown immunoregulatory role, showing SHKBP1 restrains CD8+ T cell differentiation and anti-tumor immunity.\",\n      \"evidence\": \"Germline and adenoviral Shkbp1 knockout with subcutaneous melanoma model, flow cytometry, and spleen histology\",\n      \"pmids\": [\"37343421\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Molecular pathway in T cells controlled by SHKBP1 not identified\",\n        \"Whether the immune phenotype reflects T cell-intrinsic function not resolved\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Resolved a direct molecular mechanism: SHKBP1 binds p62 to block its oligomerization and phase separation, thereby preventing Keap1 sequestration and tuning Nrf2-dependent antioxidant responses independent of ubiquitination.\",\n      \"evidence\": \"Protein-protein interaction and domain mapping, live-cell imaging of p62 bodies, and Keap1/Nrf2 reporter assays (peer-reviewed, extended from a 2025 bioRxiv preprint)\",\n      \"pmids\": [\"41649860\", \"39896619\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether the antioxidant function explains the tumor and immune phenotypes not established\",\n        \"Role of SHKBP1's Cullin-3 adaptor/ubiquitination activity in vivo not defined\",\n        \"Structural basis of the SHKBP1–p62 interaction not determined\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How SHKBP1's distinct activities — CIN85 binding, p62/Keap1/Nrf2 regulation, AKT-driven kinase switching, and CD8+ T cell control — are mechanistically unified remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"No single mechanism links the antioxidant, EMT, and immune phenotypes\",\n        \"Substrates of the SHKBP1–Cullin-3 complex unidentified\",\n        \"Tissue- and cell-type-specific functions not delineated\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"SH3KBP1\", \"SQSTM1\", \"KEAP1\", \"CUL3\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}