{"gene":"EHBP1L1","run_date":"2026-06-09T23:54:42","timeline":{"discoveries":[{"year":2016,"finding":"EHBP1L1 directly binds GTP-loaded Rab8 and Bin1/amphiphysin II, forming a complex with dynamin at the endocytic recycling compartment (ERC). This complex is required for apical-directed transport in polarized epithelial cells; EHBP1L1- or Bin1-depleted or dynamin-inhibited small intestine organoids accumulate apical membrane proteins in lysosomes. EHBP1L1-deficient mice display truncated and sparse microvilli, confirming its role in maintaining the apical plasma membrane.","method":"Biochemical pulldown, Co-IP, overexpression and knockdown in organoids, EHBP1L1-knockout mouse analysis, immunofluorescence","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal biochemical binding assays, loss-of-function in organoids with specific phenotypic readout, and in vivo knockout with defined morphological phenotype; multiple orthogonal methods in one study","pmids":["26833786"],"is_preprint":false},{"year":2018,"finding":"EHBP1L1 acts as a downstream effector of LRRK2-phosphorylated Rab8 and Rab10 on stressed lysosomes. LRRK2-mediated phosphorylation of Rab8/Rab10 recruits EHBP1L1 (and EHBP1) to overloaded lysosomes, where they mediate suppression of lysosomal enlargement and promotion of lysosomal secretion.","method":"Family-wide Rab GTPase screening, phosphorylation-dependent recruitment assays, lysosomal stress model (chloroquine), LRRK2-deficient mouse renal tubule analysis, immunofluorescence","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — systematic Rab screen plus phosphorylation-dependent localization experiments plus in vivo validation with LRRK2-KO mice; multiple orthogonal methods","pmids":["30209220"],"is_preprint":false},{"year":2020,"finding":"LRRK2-mediated phosphorylation of Rab10 blocks EHBP1L1-mediated recycling tubules and cargo turnover at macropinosomes in macrophages. EHBP1L1 overexpression competitively inhibits LRRK2 phosphorylation of Rab10 and promotes cargo recycling, mimicking LRRK2 kinase inhibition. Thus EHBP1L1 functions as an effector of non-phosphorylated (GTP-bound) Rab10 to drive fast vesicle recycling from macropinosomes.","method":"Overexpression competition assay, LRRK2 kinase inhibition, Rab10 knockdown, live-cell imaging of recycling tubules in mouse and human primary macrophages/dendritic cells/microglia-like cells","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — mechanistic competition assay with kinase inhibition and KD, live imaging, primary cells from multiple species; multiple orthogonal methods in one study","pmids":["32853409"],"is_preprint":false},{"year":2023,"finding":"EHBP1L1 interacts with and stabilizes JAK1 protein in renal cell carcinoma cells. By competing with SOCS1, EHBP1L1 protects JAK1 from proteasomal degradation, elevating JAK1 levels and activating JAK1/STAT1/PD-L1 signaling to create an immunosuppressive tumor microenvironment. EHBP1L1 depletion reduces JAK1 protein, PD-L1 surface expression, and enhances CD8+ T cell-mediated anti-tumor immunity.","method":"Co-immunoprecipitation (EHBP1L1–JAK1 interaction), competitive binding assay with SOCS1, proteasomal degradation assay, EHBP1L1 knockdown/knockout with functional immune readouts, PDX models","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP for interaction plus proteasomal degradation assay plus in vivo PDX validation, but from a single lab without independent replication","pmids":["36775874"],"is_preprint":false},{"year":2023,"finding":"EHBP1L1 is required for nuclear polarization during erythroblast enucleation. Ehbp1l1-/- mice show impaired nuclear polarization before enucleation, leading to decreased enucleation efficiency, stomatocytic erythrocyte morphology with dehydration, and early postnatal anemic lethality. EHBP1L1 interactors Rab10, Bin1, and dynamin are also involved in this process, extending the Rab8/10-EHBP1L1-Bin1-dynamin axis to erythropoiesis. EHBP1L1 loss also causes mislocalization of nuclei and mitochondria in skeletal muscle cells, phenocopying centronuclear myopathy.","method":"Ehbp1l1-knockout mouse analysis, immunofluorescence for nuclear/organelle positioning, erythrocyte morphology and dehydration assays","journal":"Blood advances","confidence":"High","confidence_rationale":"Tier 2 / Strong — complete knockout mouse with multiple cellular phenotypes (erythroblasts and muscle), mechanistic pathway placement via known interactors, orthogonal phenotypic readouts","pmids":["37042948"],"is_preprint":false},{"year":2023,"finding":"EHBP1L1 and its newly identified interactors CD2AP and CIN85 negatively regulate primary cilia length via actin network remodeling around the basal body. EHBP1L1 is localized to the ciliary sheath and is required for CD2AP/CIN85 localization there. Depletion of EHBP1L1 or CD2AP/CIN85 causes cilia elongation and actin nucleation/branching defects at the ciliary base. A CD2AP/CIN85-binding-deficient EHBP1L1 mutant fails to rescue these phenotypes, demonstrating that the EHBP1L1–CD2AP/CIN85 interaction is mechanistically required.","method":"Immunofluorescence microscopy, siRNA knockdown, rescue with WT vs. binding-deficient EHBP1L1 mutant, cilia length measurements, actin organization assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with specific ciliary phenotype, domain-specific mutant rescue, localization experiments with functional consequence; multiple orthogonal methods in one study","pmids":["36754282"],"is_preprint":false},{"year":2023,"finding":"In colorectal cancer cells, PD-L1 membrane distribution is mediated by the membrane transport complex Rab8/EHBP1L1, placing EHBP1L1 in a PCSK6/ACVR1B-p300-Rab8/EHBP1L1-PD-L1 signaling axis that controls PD-L1 surface levels.","method":"Single-cell sequencing, co-immunoprecipitation, functional rescue/inhibition in tumor cell lines and mouse models","journal":"Cell death and differentiation","confidence":"Low","confidence_rationale":"Tier 3 / Weak — EHBP1L1's role in this axis is inferred from the abstract with limited mechanistic detail specific to EHBP1L1; single study, single lab","pmids":["41975069"],"is_preprint":false},{"year":2010,"finding":"EHBP1L1 (also known as Tangerin) contains a singlet calponin homology (CH) domain capable of actin binding, placing it in the class of multidomain proteins that link actin cytoskeletal organization to vesicular trafficking.","method":"Bioinformatic domain analysis (SMART program) and comparative sequence analysis of human singlet CH-domain proteins","journal":"Molecular biology reports","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational/sequence-based domain assignment only, no direct experimental actin-binding assay described","pmids":["19459066"],"is_preprint":false},{"year":2020,"finding":"EHBP1L1 was identified as a TBC1D1-interacting protein in C2C12 myotubes via quantitative proteomics. The interaction occurs through the phosphotyrosine binding (PTB) domains of TBC1D1 and is not regulated by AMPK activation.","method":"Unbiased quantitative proteomics Co-IP in C2C12 myotubes, AMPK activation experiments","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — unbiased MS-based interactomics with AMPK-regulated controls; single lab but with quantitative orthogonal validation","pmids":["33087848"],"is_preprint":false},{"year":2025,"finding":"S1443 phosphorylation on EHBP1L1 is insulin-responsive in skeletal muscle of aged rats of both sexes, and this phosphosite is associated with insulin-stimulated glucose uptake specifically in females, suggesting EHBP1L1 participates in insulin/GLUT4 trafficking regulation via phosphorylation.","method":"Deep phosphoproteomic profiling of rat skeletal muscle with insulin stimulation and calorie restriction interventions","journal":"The journals of gerontology. Series A, Biological sciences and medical sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single phosphoproteomic study without direct mechanistic follow-up on EHBP1L1 function; correlative association between phosphosite and glucose uptake","pmids":["41105193"],"is_preprint":false},{"year":2022,"finding":"EHBP1L1 is expressed in rat primary and secondary spermatocytes and localizes with actin in the perinuclear cytoplasm adjacent to the acrosomal and Golgi regions of spermatids during acrosome biogenesis, consistent with a role in vesicular trafficking during this process.","method":"Western blot, immunofluorescence, in situ hybridization, RT-PCR on enriched testicular cell fractions","journal":"Biomedicines","confidence":"Low","confidence_rationale":"Tier 3 / Weak — localization data only, no functional loss-of-function experiment; single lab, no mechanistic causal demonstration","pmids":["35052860"],"is_preprint":false}],"current_model":"EHBP1L1 is an adapter protein that directly binds active (GTP-loaded) Rab8 and Rab10 through its bMERB domain and actin via its CH domain; it bridges these Rab GTPases to the Bin1–dynamin membrane-tubulation machinery at the endocytic recycling compartment (ERC) to drive apical-directed vesicle transport in polarized epithelial cells, lysosomal homeostasis downstream of LRRK2-phosphorylated Rab8/10, fast recycling of macropinosomal cargo in immune cells, nuclear polarization during erythroblast enucleation, and ciliary length control via recruitment of CD2AP/CIN85 and actin remodeling at the basal body; additionally, EHBP1L1 stabilizes JAK1 by competing with SOCS1 to regulate JAK1/STAT1/PD-L1 signaling in renal cell carcinoma."},"narrative":{"mechanistic_narrative":"EHBP1L1 is a multidomain adapter that couples Rab GTPase signaling to actin and the membrane-tubulation machinery to direct polarized vesicular transport [PMID:26833786]. It directly binds GTP-loaded Rab8 and Bin1/amphiphysin II to form a complex with dynamin at the endocytic recycling compartment, and this Rab8/10–EHBP1L1–Bin1–dynamin axis is required for apical-directed transport in polarized epithelial cells; its loss in mice produces truncated, sparse microvilli and missorting of apical membrane proteins to lysosomes [PMID:26833786]. The same axis operates downstream of LRRK2: phosphorylation of Rab8 and Rab10 recruits EHBP1L1 to stressed lysosomes to suppress lysosomal enlargement and promote secretion [PMID:30209220], whereas at macropinosomes EHBP1L1 acts as an effector of non-phosphorylated GTP-bound Rab10 to drive fast recycling tubule formation and cargo turnover, a function that LRRK2 phosphorylation of Rab10 antagonizes [PMID:32853409]. Beyond trafficking, EHBP1L1 controls organelle and nuclear positioning: it is required for nuclear polarization during erythroblast enucleation, with its loss causing stomatocytic erythrocytes and anemic lethality, and for correct nuclear/mitochondrial positioning in skeletal muscle [PMID:37042948], and it negatively regulates primary cilium length by localizing CD2AP/CIN85 to the ciliary sheath and remodeling basal-body actin [PMID:36754282]. In renal cell carcinoma, EHBP1L1 stabilizes JAK1 by competing with SOCS1, thereby sustaining JAK1/STAT1/PD-L1 signaling and an immunosuppressive microenvironment [PMID:36775874].","teleology":[{"year":2016,"claim":"Established EHBP1L1 as the molecular bridge linking active Rab8 to the Bin1–dynamin tubulation machinery, defining its core role in apical-directed membrane transport.","evidence":"Biochemical pulldown/Co-IP, knockdown in intestinal organoids, and EHBP1L1-knockout mouse morphology","pmids":["26833786"],"confidence":"High","gaps":["Structural basis of the Rab8/Bin1/dynamin assembly not resolved","Whether Rab10 substitutes for Rab8 in this complex not tested here"]},{"year":2018,"claim":"Placed EHBP1L1 downstream of LRRK2 signaling, showing phosphorylated Rab8/Rab10 recruit it to stressed lysosomes to control lysosomal size and secretion.","evidence":"Family-wide Rab screen, phosphorylation-dependent recruitment assays, chloroquine lysosomal stress, LRRK2-KO mouse renal tubules","pmids":["30209220"],"confidence":"High","gaps":["Direct effector mechanism at the lysosome membrane not defined","Relationship to the ERC trafficking role unclear"]},{"year":2020,"claim":"Resolved how Rab10 phosphorylation state gates EHBP1L1 activity, showing it is an effector of non-phosphorylated GTP-Rab10 that drives fast macropinosomal recycling antagonized by LRRK2.","evidence":"Overexpression competition assay, LRRK2 kinase inhibition, Rab10 knockdown, live-cell imaging in primary macrophages/dendritic cells/microglia","pmids":["32853409"],"confidence":"High","gaps":["Quantitative competition between EHBP1L1 binding and LRRK2 phosphorylation not measured","Cargo specificity of recycling tubules undefined"]},{"year":2020,"claim":"Identified EHBP1L1 as a TBC1D1 interactor via its PTB domains, linking it to a Rab-GAP in myotubes independent of AMPK.","evidence":"Unbiased quantitative proteomics Co-IP in C2C12 myotubes with AMPK activation controls","pmids":["33087848"],"confidence":"Medium","gaps":["Functional consequence of the TBC1D1 interaction not tested","No reciprocal validation or in vivo confirmation"]},{"year":2023,"claim":"Extended the Rab/Bin1/dynamin axis to nuclear and organelle positioning, showing EHBP1L1 is required for erythroblast nuclear polarization during enucleation and for nuclear/mitochondrial positioning in muscle.","evidence":"Ehbp1l1-knockout mouse analysis, immunofluorescence of organelle positioning, erythrocyte morphology assays","pmids":["37042948"],"confidence":"High","gaps":["Mechanism connecting vesicular transport to nuclear polarization unresolved","Whether the muscle phenotype reflects the same molecular pathway untested"]},{"year":2023,"claim":"Defined a ciliary function via newly identified CD2AP/CIN85 partners, showing EHBP1L1 restrains cilium length through basal-body actin remodeling.","evidence":"siRNA knockdown, rescue with WT vs CD2AP/CIN85-binding-deficient mutant, cilia length and actin organization assays","pmids":["36754282"],"confidence":"High","gaps":["How CD2AP/CIN85 recruitment couples to actin nucleation not detailed","Relationship between ciliary and trafficking roles unclear"]},{"year":2023,"claim":"Revealed a non-trafficking role in tumor immune evasion, showing EHBP1L1 stabilizes JAK1 by competing with SOCS1 to sustain STAT1/PD-L1 signaling.","evidence":"Co-IP, SOCS1 competitive binding assay, proteasomal degradation assay, knockdown/knockout with immune readouts and PDX models","pmids":["36775874"],"confidence":"Medium","gaps":["Single lab without independent replication","Domain of EHBP1L1 mediating JAK1 binding not mapped","Connection to its Rab-effector function unknown"]},{"year":null,"claim":"How EHBP1L1's distinct activities — Rab/Bin1/dynamin trafficking, basal-body actin remodeling, and JAK1 stabilization — are partitioned across cell types and integrated by phosphorylation remains unresolved.","evidence":"No single study reconciles the trafficking, ciliary, and signaling roles","pmids":[],"confidence":"Low","gaps":["No structural model of full-length EHBP1L1 with its partners","Regulatory phosphosites (e.g. S1443) lack functional validation","Tissue-specific partner usage undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,2]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,5,7]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[1]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[5]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[5,7]}],"pathway":[{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,2]},{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[0]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,3]}],"complexes":[],"partners":["RAB8A","RAB10","BIN1","DNM2","CD2AP","SH3KBP1","JAK1","TBC1D1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N3D4","full_name":"EH domain-binding protein 1-like protein 1","aliases":[],"length_aa":1523,"mass_kda":161.9,"function":"May act as Rab effector protein and play a role in vesicle trafficking","subcellular_location":"Endosome","url":"https://www.uniprot.org/uniprotkb/Q8N3D4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EHBP1L1","classification":"Not Classified","n_dependent_lines":66,"n_total_lines":1208,"dependency_fraction":0.054635761589403975},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/EHBP1L1","total_profiled":1310},"omim":[{"mim_id":"619583","title":"EH DOMAIN-BINDING PROTEIN 1-LIKE 1; EHBP1L1","url":"https://www.omim.org/entry/619583"},{"mim_id":"612672","title":"RAS-ASSOCIATED PROTEIN RAB10; RAB10","url":"https://www.omim.org/entry/612672"},{"mim_id":"609007","title":"LEUCINE-RICH REPEAT KINASE 2; LRRK2","url":"https://www.omim.org/entry/609007"},{"mim_id":"601248","title":"BRIDGING INTEGRATOR 1; BIN1","url":"https://www.omim.org/entry/601248"},{"mim_id":"165040","title":"RAS-ASSOCIATED PROTEIN RAB8A; RAB8A","url":"https://www.omim.org/entry/165040"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"},{"location":"Plasma membrane","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"skeletal muscle","ntpm":239.6}],"url":"https://www.proteinatlas.org/search/EHBP1L1"},"hgnc":{"alias_symbol":["DKFZp762C186","TANGERIN"],"prev_symbol":[]},"alphafold":{"accession":"Q8N3D4","domains":[{"cath_id":"2.60.40.150","chopping":"2-171","consensus_level":"high","plddt":87.9184,"start":2,"end":171},{"cath_id":"1.10.418.10","chopping":"1040-1143","consensus_level":"high","plddt":87.334,"start":1040,"end":1143},{"cath_id":"-","chopping":"1352-1445_1470-1508","consensus_level":"medium","plddt":83.4855,"start":1352,"end":1508}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N3D4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N3D4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N3D4-F1-predicted_aligned_error_v6.png","plddt_mean":48.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EHBP1L1","jax_strain_url":"https://www.jax.org/strain/search?query=EHBP1L1"},"sequence":{"accession":"Q8N3D4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N3D4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N3D4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N3D4"}},"corpus_meta":[{"pmid":"30209220","id":"PMC_30209220","title":"LRRK2 and its substrate Rab GTPases are sequentially targeted onto stressed lysosomes and maintain their homeostasis.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/30209220","citation_count":249,"is_preprint":false},{"pmid":"26515236","id":"PMC_26515236","title":"A CpG-methylation-based assay to predict survival in clear cell renal cell carcinoma.","date":"2015","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/26515236","citation_count":101,"is_preprint":false},{"pmid":"24954895","id":"PMC_24954895","title":"Gene-age interactions in blood pressure regulation: a large-scale investigation with the CHARGE, Global BPgen, and ICBP Consortia.","date":"2014","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/24954895","citation_count":100,"is_preprint":false},{"pmid":"32853409","id":"PMC_32853409","title":"LRRK2 and Rab10 coordinate macropinocytosis to mediate immunological responses in phagocytes.","date":"2020","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/32853409","citation_count":76,"is_preprint":false},{"pmid":"26833786","id":"PMC_26833786","title":"EHBP1L1 coordinates Rab8 and Bin1 to regulate apical-directed transport in polarized epithelial cells.","date":"2016","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/26833786","citation_count":53,"is_preprint":false},{"pmid":"25335978","id":"PMC_25335978","title":"Comprehensive replication of the relationship between myopia-related genes and refractive errors in a large Japanese cohort.","date":"2014","source":"Investigative ophthalmology & visual science","url":"https://pubmed.ncbi.nlm.nih.gov/25335978","citation_count":47,"is_preprint":false},{"pmid":"36775874","id":"PMC_36775874","title":"EHBP1L1 Drives Immune Evasion in Renal Cell Carcinoma through Binding and Stabilizing JAK1.","date":"2023","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/36775874","citation_count":36,"is_preprint":false},{"pmid":"34645488","id":"PMC_34645488","title":"Lethal variants in humans: lessons learned from a large molecular autopsy cohort.","date":"2021","source":"Genome medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34645488","citation_count":27,"is_preprint":false},{"pmid":"33087848","id":"PMC_33087848","title":"TBC1D1 interacting proteins, VPS13A and VPS13C, regulate GLUT4 homeostasis in C2C12 myotubes.","date":"2020","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/33087848","citation_count":16,"is_preprint":false},{"pmid":"37042948","id":"PMC_37042948","title":"EHBP1L1, an apicobasal polarity regulator, is critical for nuclear polarization during enucleation of erythroblasts.","date":"2023","source":"Blood advances","url":"https://pubmed.ncbi.nlm.nih.gov/37042948","citation_count":12,"is_preprint":false},{"pmid":"35052860","id":"PMC_35052860","title":"Differential Expression and Localization of EHBP1L1 during the First Wave of Rat Spermatogenesis Suggest Its Involvement in Acrosome Biogenesis.","date":"2022","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/35052860","citation_count":10,"is_preprint":false},{"pmid":"39198424","id":"PMC_39198424","title":"Genome-wide DNA methylation profiling in blood reveals epigenetic signature of incident acute coronary syndrome.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/39198424","citation_count":10,"is_preprint":false},{"pmid":"19459066","id":"PMC_19459066","title":"Singlet CH domain containing human multidomain proteins: an inventory.","date":"2009","source":"Molecular biology 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This complex is required for apical-directed transport in polarized epithelial cells; EHBP1L1- or Bin1-depleted or dynamin-inhibited small intestine organoids accumulate apical membrane proteins in lysosomes. EHBP1L1-deficient mice display truncated and sparse microvilli, confirming its role in maintaining the apical plasma membrane.\",\n      \"method\": \"Biochemical pulldown, Co-IP, overexpression and knockdown in organoids, EHBP1L1-knockout mouse analysis, immunofluorescence\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal biochemical binding assays, loss-of-function in organoids with specific phenotypic readout, and in vivo knockout with defined morphological phenotype; multiple orthogonal methods in one study\",\n      \"pmids\": [\"26833786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"EHBP1L1 acts as a downstream effector of LRRK2-phosphorylated Rab8 and Rab10 on stressed lysosomes. LRRK2-mediated phosphorylation of Rab8/Rab10 recruits EHBP1L1 (and EHBP1) to overloaded lysosomes, where they mediate suppression of lysosomal enlargement and promotion of lysosomal secretion.\",\n      \"method\": \"Family-wide Rab GTPase screening, phosphorylation-dependent recruitment assays, lysosomal stress model (chloroquine), LRRK2-deficient mouse renal tubule analysis, immunofluorescence\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — systematic Rab screen plus phosphorylation-dependent localization experiments plus in vivo validation with LRRK2-KO mice; multiple orthogonal methods\",\n      \"pmids\": [\"30209220\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"LRRK2-mediated phosphorylation of Rab10 blocks EHBP1L1-mediated recycling tubules and cargo turnover at macropinosomes in macrophages. EHBP1L1 overexpression competitively inhibits LRRK2 phosphorylation of Rab10 and promotes cargo recycling, mimicking LRRK2 kinase inhibition. Thus EHBP1L1 functions as an effector of non-phosphorylated (GTP-bound) Rab10 to drive fast vesicle recycling from macropinosomes.\",\n      \"method\": \"Overexpression competition assay, LRRK2 kinase inhibition, Rab10 knockdown, live-cell imaging of recycling tubules in mouse and human primary macrophages/dendritic cells/microglia-like cells\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — mechanistic competition assay with kinase inhibition and KD, live imaging, primary cells from multiple species; multiple orthogonal methods in one study\",\n      \"pmids\": [\"32853409\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EHBP1L1 interacts with and stabilizes JAK1 protein in renal cell carcinoma cells. By competing with SOCS1, EHBP1L1 protects JAK1 from proteasomal degradation, elevating JAK1 levels and activating JAK1/STAT1/PD-L1 signaling to create an immunosuppressive tumor microenvironment. EHBP1L1 depletion reduces JAK1 protein, PD-L1 surface expression, and enhances CD8+ T cell-mediated anti-tumor immunity.\",\n      \"method\": \"Co-immunoprecipitation (EHBP1L1–JAK1 interaction), competitive binding assay with SOCS1, proteasomal degradation assay, EHBP1L1 knockdown/knockout with functional immune readouts, PDX models\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP for interaction plus proteasomal degradation assay plus in vivo PDX validation, but from a single lab without independent replication\",\n      \"pmids\": [\"36775874\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EHBP1L1 is required for nuclear polarization during erythroblast enucleation. Ehbp1l1-/- mice show impaired nuclear polarization before enucleation, leading to decreased enucleation efficiency, stomatocytic erythrocyte morphology with dehydration, and early postnatal anemic lethality. EHBP1L1 interactors Rab10, Bin1, and dynamin are also involved in this process, extending the Rab8/10-EHBP1L1-Bin1-dynamin axis to erythropoiesis. EHBP1L1 loss also causes mislocalization of nuclei and mitochondria in skeletal muscle cells, phenocopying centronuclear myopathy.\",\n      \"method\": \"Ehbp1l1-knockout mouse analysis, immunofluorescence for nuclear/organelle positioning, erythrocyte morphology and dehydration assays\",\n      \"journal\": \"Blood advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — complete knockout mouse with multiple cellular phenotypes (erythroblasts and muscle), mechanistic pathway placement via known interactors, orthogonal phenotypic readouts\",\n      \"pmids\": [\"37042948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"EHBP1L1 and its newly identified interactors CD2AP and CIN85 negatively regulate primary cilia length via actin network remodeling around the basal body. EHBP1L1 is localized to the ciliary sheath and is required for CD2AP/CIN85 localization there. Depletion of EHBP1L1 or CD2AP/CIN85 causes cilia elongation and actin nucleation/branching defects at the ciliary base. A CD2AP/CIN85-binding-deficient EHBP1L1 mutant fails to rescue these phenotypes, demonstrating that the EHBP1L1–CD2AP/CIN85 interaction is mechanistically required.\",\n      \"method\": \"Immunofluorescence microscopy, siRNA knockdown, rescue with WT vs. binding-deficient EHBP1L1 mutant, cilia length measurements, actin organization assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with specific ciliary phenotype, domain-specific mutant rescue, localization experiments with functional consequence; multiple orthogonal methods in one study\",\n      \"pmids\": [\"36754282\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In colorectal cancer cells, PD-L1 membrane distribution is mediated by the membrane transport complex Rab8/EHBP1L1, placing EHBP1L1 in a PCSK6/ACVR1B-p300-Rab8/EHBP1L1-PD-L1 signaling axis that controls PD-L1 surface levels.\",\n      \"method\": \"Single-cell sequencing, co-immunoprecipitation, functional rescue/inhibition in tumor cell lines and mouse models\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — EHBP1L1's role in this axis is inferred from the abstract with limited mechanistic detail specific to EHBP1L1; single study, single lab\",\n      \"pmids\": [\"41975069\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"EHBP1L1 (also known as Tangerin) contains a singlet calponin homology (CH) domain capable of actin binding, placing it in the class of multidomain proteins that link actin cytoskeletal organization to vesicular trafficking.\",\n      \"method\": \"Bioinformatic domain analysis (SMART program) and comparative sequence analysis of human singlet CH-domain proteins\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational/sequence-based domain assignment only, no direct experimental actin-binding assay described\",\n      \"pmids\": [\"19459066\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EHBP1L1 was identified as a TBC1D1-interacting protein in C2C12 myotubes via quantitative proteomics. The interaction occurs through the phosphotyrosine binding (PTB) domains of TBC1D1 and is not regulated by AMPK activation.\",\n      \"method\": \"Unbiased quantitative proteomics Co-IP in C2C12 myotubes, AMPK activation experiments\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — unbiased MS-based interactomics with AMPK-regulated controls; single lab but with quantitative orthogonal validation\",\n      \"pmids\": [\"33087848\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"S1443 phosphorylation on EHBP1L1 is insulin-responsive in skeletal muscle of aged rats of both sexes, and this phosphosite is associated with insulin-stimulated glucose uptake specifically in females, suggesting EHBP1L1 participates in insulin/GLUT4 trafficking regulation via phosphorylation.\",\n      \"method\": \"Deep phosphoproteomic profiling of rat skeletal muscle with insulin stimulation and calorie restriction interventions\",\n      \"journal\": \"The journals of gerontology. Series A, Biological sciences and medical sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single phosphoproteomic study without direct mechanistic follow-up on EHBP1L1 function; correlative association between phosphosite and glucose uptake\",\n      \"pmids\": [\"41105193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"EHBP1L1 is expressed in rat primary and secondary spermatocytes and localizes with actin in the perinuclear cytoplasm adjacent to the acrosomal and Golgi regions of spermatids during acrosome biogenesis, consistent with a role in vesicular trafficking during this process.\",\n      \"method\": \"Western blot, immunofluorescence, in situ hybridization, RT-PCR on enriched testicular cell fractions\",\n      \"journal\": \"Biomedicines\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — localization data only, no functional loss-of-function experiment; single lab, no mechanistic causal demonstration\",\n      \"pmids\": [\"35052860\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EHBP1L1 is an adapter protein that directly binds active (GTP-loaded) Rab8 and Rab10 through its bMERB domain and actin via its CH domain; it bridges these Rab GTPases to the Bin1–dynamin membrane-tubulation machinery at the endocytic recycling compartment (ERC) to drive apical-directed vesicle transport in polarized epithelial cells, lysosomal homeostasis downstream of LRRK2-phosphorylated Rab8/10, fast recycling of macropinosomal cargo in immune cells, nuclear polarization during erythroblast enucleation, and ciliary length control via recruitment of CD2AP/CIN85 and actin remodeling at the basal body; additionally, EHBP1L1 stabilizes JAK1 by competing with SOCS1 to regulate JAK1/STAT1/PD-L1 signaling in renal cell carcinoma.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EHBP1L1 is a multidomain adapter that couples Rab GTPase signaling to actin and the membrane-tubulation machinery to direct polarized vesicular transport [#0]. It directly binds GTP-loaded Rab8 and Bin1/amphiphysin II to form a complex with dynamin at the endocytic recycling compartment, and this Rab8/10–EHBP1L1–Bin1–dynamin axis is required for apical-directed transport in polarized epithelial cells; its loss in mice produces truncated, sparse microvilli and missorting of apical membrane proteins to lysosomes [#0]. The same axis operates downstream of LRRK2: phosphorylation of Rab8 and Rab10 recruits EHBP1L1 to stressed lysosomes to suppress lysosomal enlargement and promote secretion [#1], whereas at macropinosomes EHBP1L1 acts as an effector of non-phosphorylated GTP-bound Rab10 to drive fast recycling tubule formation and cargo turnover, a function that LRRK2 phosphorylation of Rab10 antagonizes [#2]. Beyond trafficking, EHBP1L1 controls organelle and nuclear positioning: it is required for nuclear polarization during erythroblast enucleation, with its loss causing stomatocytic erythrocytes and anemic lethality, and for correct nuclear/mitochondrial positioning in skeletal muscle [#4], and it negatively regulates primary cilium length by localizing CD2AP/CIN85 to the ciliary sheath and remodeling basal-body actin [#5]. In renal cell carcinoma, EHBP1L1 stabilizes JAK1 by competing with SOCS1, thereby sustaining JAK1/STAT1/PD-L1 signaling and an immunosuppressive microenvironment [#3].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Established EHBP1L1 as the molecular bridge linking active Rab8 to the Bin1–dynamin tubulation machinery, defining its core role in apical-directed membrane transport.\",\n      \"evidence\": \"Biochemical pulldown/Co-IP, knockdown in intestinal organoids, and EHBP1L1-knockout mouse morphology\",\n      \"pmids\": [\"26833786\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of the Rab8/Bin1/dynamin assembly not resolved\", \"Whether Rab10 substitutes for Rab8 in this complex not tested here\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed EHBP1L1 downstream of LRRK2 signaling, showing phosphorylated Rab8/Rab10 recruit it to stressed lysosomes to control lysosomal size and secretion.\",\n      \"evidence\": \"Family-wide Rab screen, phosphorylation-dependent recruitment assays, chloroquine lysosomal stress, LRRK2-KO mouse renal tubules\",\n      \"pmids\": [\"30209220\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct effector mechanism at the lysosome membrane not defined\", \"Relationship to the ERC trafficking role unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Resolved how Rab10 phosphorylation state gates EHBP1L1 activity, showing it is an effector of non-phosphorylated GTP-Rab10 that drives fast macropinosomal recycling antagonized by LRRK2.\",\n      \"evidence\": \"Overexpression competition assay, LRRK2 kinase inhibition, Rab10 knockdown, live-cell imaging in primary macrophages/dendritic cells/microglia\",\n      \"pmids\": [\"32853409\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative competition between EHBP1L1 binding and LRRK2 phosphorylation not measured\", \"Cargo specificity of recycling tubules undefined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified EHBP1L1 as a TBC1D1 interactor via its PTB domains, linking it to a Rab-GAP in myotubes independent of AMPK.\",\n      \"evidence\": \"Unbiased quantitative proteomics Co-IP in C2C12 myotubes with AMPK activation controls\",\n      \"pmids\": [\"33087848\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the TBC1D1 interaction not tested\", \"No reciprocal validation or in vivo confirmation\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended the Rab/Bin1/dynamin axis to nuclear and organelle positioning, showing EHBP1L1 is required for erythroblast nuclear polarization during enucleation and for nuclear/mitochondrial positioning in muscle.\",\n      \"evidence\": \"Ehbp1l1-knockout mouse analysis, immunofluorescence of organelle positioning, erythrocyte morphology assays\",\n      \"pmids\": [\"37042948\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism connecting vesicular transport to nuclear polarization unresolved\", \"Whether the muscle phenotype reflects the same molecular pathway untested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined a ciliary function via newly identified CD2AP/CIN85 partners, showing EHBP1L1 restrains cilium length through basal-body actin remodeling.\",\n      \"evidence\": \"siRNA knockdown, rescue with WT vs CD2AP/CIN85-binding-deficient mutant, cilia length and actin organization assays\",\n      \"pmids\": [\"36754282\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CD2AP/CIN85 recruitment couples to actin nucleation not detailed\", \"Relationship between ciliary and trafficking roles unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Revealed a non-trafficking role in tumor immune evasion, showing EHBP1L1 stabilizes JAK1 by competing with SOCS1 to sustain STAT1/PD-L1 signaling.\",\n      \"evidence\": \"Co-IP, SOCS1 competitive binding assay, proteasomal degradation assay, knockdown/knockout with immune readouts and PDX models\",\n      \"pmids\": [\"36775874\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Single lab without independent replication\", \"Domain of EHBP1L1 mediating JAK1 binding not mapped\", \"Connection to its Rab-effector function unknown\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How EHBP1L1's distinct activities — Rab/Bin1/dynamin trafficking, basal-body actin remodeling, and JAK1 stabilization — are partitioned across cell types and integrated by phosphorylation remains unresolved.\",\n      \"evidence\": \"No single study reconciles the trafficking, ciliary, and signaling roles\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of full-length EHBP1L1 with its partners\", \"Regulatory phosphosites (e.g. S1443) lack functional validation\", \"Tissue-specific partner usage undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 2]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 5, 7]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [5, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 3]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RAB8A\", \"RAB10\", \"BIN1\", \"DNM2\", \"CD2AP\", \"SH3KBP1\", \"JAK1\", \"TBC1D1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}