{"gene":"EVI5","run_date":"2026-06-09T23:54:43","timeline":{"discoveries":[{"year":2006,"finding":"Evi5 stabilizes Emi1 (an APC/C inhibitor) by binding adjacent to Emi1's DSGxxS degron, blocking both degron phosphorylation by Polo-like kinases and subsequent betaTrCP binding, thereby antagonizing SCF(betaTrCP)-dependent Emi1 ubiquitination and destruction. Evi5 protein accumulates in early G1 following Plk1 destruction and is itself degraded in a Plk1- and ubiquitin-dependent manner in early mitosis. Ablation of Evi5 causes precocious Emi1 degradation, premature APC/C activation, cyclin destruction, centrosome overduplication, and mitotic catastrophe.","method":"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, cell-cycle imaging, kinase assays","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal biochemical methods (binding assay, ubiquitination assay, mutagenesis of degron, kinase assays) plus cellular phenotypic rescue in a single rigorous study","pmids":["16439210"],"is_preprint":false},{"year":2007,"finding":"Evi5 binds Rab11a and Rab11b in a GTP-dependent manner (identified by yeast two-hybrid, co-immunoprecipitation, and Biacore surface plasmon resonance), but displays no GTPase-activating activity toward Rab11 in vitro. Evi5 colocalizes with Rab11 in vivo; Rab11 effector proteins (e.g., FIP3) compete with Evi5 for Rab11 binding. Ablation of Evi5 by RNAi causes mislocalization of FIP3 at the abscission site during cytokinesis.","method":"Yeast two-hybrid, co-immunoprecipitation, Biacore SPR, in vitro GAP assay, RNAi knockdown, confocal microscopy","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal binding methods (Y2H, Co-IP, SPR), in vitro GAP assay, and cellular phenotype with RNAi in a single study","pmids":["17229837"],"is_preprint":false},{"year":2006,"finding":"EVI5's TBC domain functions as a GTPase-activating protein (GAP) for Rab11: EVI5 preferentially binds the GTP-bound form of Rab11 and stimulates Rab11 GTPase activity in a GAP activity assay, as demonstrated by mass spectrometry-identified complex formation and a specific Rab-binding/GAP assay.","method":"Linear ion trap mass spectrometry (protein complex identification), GTP-form-selective Rab-binding assay, in vitro GAP activity assay","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro GAP assay plus MS-based complex identification, single lab but two orthogonal methods; note this finding contradicts Westlake et al. 2007 which found no GAP activity, but this paper reports positive GAP stimulation","pmids":["17099728"],"is_preprint":false},{"year":2005,"finding":"EVI5 localizes to pericentriolar material (centrosome) in interphase cells and exists in protein complexes with both alpha- and gamma-tubulin, with both interactions mapping to the N-terminal region of EVI5. EVI5 can form dimers as shown by FPLC analysis.","method":"Confocal immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, FPLC gel filtration","journal":"Genomics","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal Co-IP and GST pull-down plus localization, single lab","pmids":["16033705"],"is_preprint":false},{"year":2006,"finding":"EVI5 has a dynamic mitotic distribution: it associates with the mitotic spindle through anaphase and localizes to the midzone and midbody until completion of cytokinesis. EVI5 undergoes phosphorylation in early mitosis and proteolytic cleavage during late mitosis/cytokinesis. EVI5 co-immunoprecipitates and co-purifies (GST pull-down) with the chromosomal passenger complex (INCENP, Aurora B kinase, survivin). siRNA knockdown of EVI5 produces a multinucleate phenotype, establishing an essential role in cytokinesis completion.","method":"Immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, siRNA knockdown, Western blot for phosphorylation/cleavage","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal Co-IP, GST pull-down, and siRNA phenotype, single lab with multiple orthogonal methods","pmids":["16764853"],"is_preprint":false},{"year":2012,"finding":"Drosophila Evi5 promotes collective border cell migration through its Rab-GAP activity directed at Rab11. Both loss and gain of Evi5 function block border cell migration by disrupting Rab11-dependent polarization of active guidance receptors. The TBC/Rab-GAP domain is required for this function.","method":"Drosophila genetic screen for Rab-GAP proteins, loss-of-function and gain-of-function genetics, in vivo imaging of border cell migration, Rab11 activity reporters","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic epistasis with domain-requirement validation and in vivo imaging in Drosophila ortholog, replicates the Rab11-GAP function","pmids":["22778279"],"is_preprint":false},{"year":1998,"finding":"The human EVI5 gene (NB4S) encodes an 810 amino acid protein with a TBC1 box motif (homologous to genes involved in cell growth/differentiation) and C-terminal coiled-coil domains. It is disrupted by a constitutional t(1;10)(p22;q21) translocation in a neuroblastoma patient, generating a fusion transcript that combines the TBC1 motif of NB4S with a polyadenylation signal from TRNG10, potentially producing a truncated oncogenic protein.","method":"Molecular cloning of translocation breakpoints, sequence analysis, Northern blot expression analysis","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — direct molecular cloning and sequencing of the translocation breakpoint, single lab","pmids":["9618176"],"is_preprint":false},{"year":2009,"finding":"EVI5 overexpression prevents Runx1-deficient hematopoietic stem cell (HSC) exhaustion in mice, identified by retroviral insertional mutagenesis as a cooperating genetic alteration in leukemia development. EVI5 activation maintains pre-leukemic stem cell populations in the context of Runx1 loss.","method":"Retroviral insertional mutagenesis screen, conditional Runx1 knockout mouse model, retroviral EVI5 overexpression rescue experiment","journal":"Blood","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with in vivo rescue experiment, single lab","pmids":["20008790"],"is_preprint":false},{"year":2020,"finding":"Evi5 interacts with c-Myc and antagonizes FBXW7 E3 ligase-mediated ubiquitination and degradation of c-Myc protein, thereby stabilizing c-Myc and promoting its transcriptional activity in laryngeal squamous cell carcinoma cells. CRISPR-mediated Evi5 knockout decreases c-Myc levels, causes G1 arrest, and reduces tumor growth in vivo.","method":"Co-immunoprecipitation, CRISPR knockout, luciferase transcriptional reporter assay, flow cytometry, xenograft mouse model","journal":"Cancer cell international","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — Co-IP for interaction, CRISPR KO with multiple phenotypic readouts, single lab","pmids":["32047362"],"is_preprint":false},{"year":2020,"finding":"EVI5 interacts with TGF-β receptor I and TGF-β receptor II (shown by co-immunoprecipitation), and the miR-486-5p–EVI5 axis affects NSCLC migration and invasion through the TGF-β/Smad signaling pathway. EVI5 also interacts with Emi1 in NSCLC cells.","method":"Co-immunoprecipitation, siRNA knockdown, wound healing/transwell invasion assays, xenograft mouse model","journal":"Journal of experimental & clinical cancer research : CR","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP for TGFβR interaction, single lab, no mutagenesis or reconstitution to confirm mechanistic link","pmids":["32393392"],"is_preprint":false},{"year":2015,"finding":"An exonic SNP in EVI5 associated with multiple sclerosis risk induces changes in superficial hydrophobicity patterns of the EVI5 coiled-coil domain, altering the EVI5 interactome. Immunoprecipitation followed by mass spectrometry of wild-type vs. risk-variant EVI5 revealed disease-specific interactors linked to lipid metabolism, including a novel interaction with sphingosine 1-phosphate lyase (SGPL1) exclusive to the risk variant.","method":"Fine mapping/meta-analysis, co-immunoprecipitation followed by mass spectrometry, structural modeling of coiled-coil domain","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — IP-MS interactome comparison with functional variant, single lab, novel interaction identified by two orthogonal approaches (IP-MS and structural analysis)","pmids":["26433934"],"is_preprint":false},{"year":2024,"finding":"Drosophila Evi5 is required for intracellular iron trafficking: Evi5 depletion in the prothoracic gland disrupts vesicle morphology, blocks endosome recycling, and impairs transferrin-1 trafficking, reducing cellular iron and impairing heme synthesis. Evi5 physically interacts with ferritin, and ferritin injection rescues developmental delays caused by Evi5 depletion.","method":"RNAi depletion in Drosophila prothoracic gland, electron microscopy of vesicle morphology, ferritin co-immunoprecipitation/physical interaction assay, ferritin injection rescue experiment","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (RNAi, EM, physical interaction, rescue), single lab, Drosophila ortholog","pmids":["38744835"],"is_preprint":false},{"year":2025,"finding":"EVI5 interacts with Rab11 and upregulates PD-L1 expression in lung adenocarcinoma cells. EVI5 knockout suppresses Rab11 and PD-L1 expression, while EVI5 overexpression promotes their expression, defining an EVI5–Rab11–PD-L1 regulatory axis.","method":"Co-immunoprecipitation, CRISPR knockout, overexpression, Western blot","journal":"Scientific reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP for interaction, single lab, no mechanistic dissection of how Rab11 connects to PD-L1","pmids":["41258404"],"is_preprint":false},{"year":2025,"finding":"EVI5 promotes lung adenocarcinoma progression through activation of the ERK1/2–c-Myc signaling axis; EVI5 knockout reduces ERK1/2 and c-Myc activity, and an ERK1/2 pathway inhibitor phenocopies EVI5 loss, validated by functional rescue experiments.","method":"CRISPR knockout, overexpression, Western blot, ERK1/2 inhibitor functional rescue, immunofluorescence, qRT-PCR","journal":"Current cancer drug targets","confidence":"Low","confidence_rationale":"Tier 3 / Weak — genetic epistasis via inhibitor rescue, single lab, no direct biochemical interaction between EVI5 and ERK1/2 demonstrated","pmids":["40525449"],"is_preprint":false},{"year":2022,"finding":"Evi5 is required for Xenopus limb and tail regeneration: morpholino-mediated Evi5 knockdown reduces blastema cell proliferation, causes apoptosis, and blocks blastema formation. RNA-seq reveals Evi5 knockdown downregulates PDGFα and TGFβ signaling pathways as well as lysine demethylases Kdm6b and Kdm7a, which are themselves required for limb regeneration.","method":"Morpholino antisense knockdown in Xenopus tadpoles, RNA-sequencing, proliferation and apoptosis assays, Kdm6b/Kdm7a knockdown epistasis","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — morpholino knockdown with RNA-seq and epistasis experiments in Xenopus ortholog, single lab","pmids":["36605717"],"is_preprint":false}],"current_model":"EVI5 is a TBC-domain-containing protein that functions as a Rab11 GTPase-activating protein (or GTP-dependent Rab11 binding partner) to regulate endosomal recycling, cytokinesis, and cell migration; it also acts as a cell cycle regulator by stabilizing Emi1 (blocking SCF(betaTrCP)-mediated ubiquitination) to control APC/C activity and cyclin accumulation, associates with the Aurora B/INCENP/survivin chromosomal passenger complex at the midbody, localizes to centrosomes via interactions with alpha- and gamma-tubulin, and can stabilize oncoproteins such as c-Myc by antagonizing FBXW7-dependent degradation."},"narrative":{"mechanistic_narrative":"EVI5 is a TBC-domain protein that integrates membrane-trafficking control with cell-cycle regulation, acting at the interface of Rab11-dependent endosomal recycling and the mitotic machinery [PMID:17229837, PMID:16439210]. Through its TBC/Rab-GAP domain EVI5 binds Rab11 selectively in its GTP-bound state and governs Rab11-dependent recycling, polarizing guidance-receptor delivery during collective cell migration [PMID:17229837, PMID:22778279]; whether it stimulates Rab11 GTP hydrolysis is reported divergently, with one study finding GAP activity [PMID:17099728] and another finding GTP-dependent binding without measurable GAP activity in vitro [PMID:17229837]. In parallel, EVI5 controls cell-cycle progression by binding adjacent to the DSGxxS degron of the APC/C inhibitor Emi1, blocking Plk1-mediated degron phosphorylation and subsequent betaTrCP recruitment, thereby stabilizing Emi1 and restraining premature APC/C activation; EVI5 itself accumulates in early G1 and is degraded in a Plk1- and ubiquitin-dependent manner in early mitosis, and its loss causes precocious Emi1 destruction, cyclin loss, centrosome overduplication, and mitotic catastrophe [PMID:16439210]. EVI5 has a dynamic mitotic distribution, localizing to centrosomal pericentriolar material via N-terminal interactions with alpha- and gamma-tubulin and tracking the spindle, midzone, and midbody, where it associates with the chromosomal passenger complex (INCENP, Aurora B, survivin) and is required for cytokinesis completion [PMID:16033705, PMID:16764853]. EVI5 was first identified as a gene (NB4S) disrupted by a constitutional translocation in neuroblastoma [PMID:9618176], and subsequent work links it to oncogenic and developmental roles: it antagonizes FBXW7-mediated ubiquitination of c-Myc to stabilize the oncoprotein [PMID:32047362], maintains pre-leukemic hematopoietic stem cells in the setting of Runx1 loss [PMID:20008790], and is required for vertebrate appendage regeneration through PDGF/TGF-beta signaling and lysine demethylases [PMID:36605717].","teleology":[{"year":1998,"claim":"Establishing the molecular identity of EVI5 — that the gene encodes a TBC1-motif, coiled-coil protein disrupted in cancer — provided the first structural framework for its later-defined GAP and scaffolding functions.","evidence":"molecular cloning and sequencing of a t(1;10) translocation breakpoint in a neuroblastoma patient, with Northern expression analysis","pmids":["9618176"],"confidence":"Medium","gaps":["Did not establish the biochemical activity of the TBC1 motif","Oncogenic consequence of the fusion transcript inferred but not functionally tested"]},{"year":2005,"claim":"Localizing EVI5 to the centrosome and showing it binds alpha- and gamma-tubulin defined a structural/scaffolding role at the pericentriolar material before its enzymatic activity was known.","evidence":"confocal immunofluorescence, reciprocal Co-IP, GST pull-down, and FPLC gel filtration","pmids":["16033705"],"confidence":"Medium","gaps":["Functional consequence of tubulin binding not tested","Single lab, no in vivo validation of centrosomal function"]},{"year":2006,"claim":"Identifying EVI5 as a stabilizer of the APC/C inhibitor Emi1 placed it directly in cell-cycle control, explaining why its loss triggers premature APC/C activation and mitotic catastrophe.","evidence":"Co-IP, in vitro ubiquitination assay, degron mutagenesis, kinase assays, siRNA knockdown with cell-cycle imaging","pmids":["16439210"],"confidence":"High","gaps":["Structural basis of degron-adjacent binding not resolved","Relationship between Emi1-stabilizing and Rab-GAP functions of the same protein not integrated"]},{"year":2006,"claim":"Tracking EVI5's mitotic distribution and its association with the chromosomal passenger complex established a midbody role essential for cytokinesis completion, complementing its earlier centrosomal localization.","evidence":"immunofluorescence across mitosis, Co-IP/GST pull-down with INCENP/Aurora B/survivin, siRNA producing a multinucleate phenotype, Western blots for phosphorylation/cleavage","pmids":["16764853"],"confidence":"Medium","gaps":["Functional significance of CPC association not dissected","Identity of the protease cleaving EVI5 in late mitosis unknown"]},{"year":2006,"claim":"Demonstrating that the EVI5 TBC domain stimulates Rab11 GTP hydrolysis assigned it a Rab11-GAP enzymatic activity, linking it to endosomal trafficking control.","evidence":"linear ion-trap MS complex identification, GTP-form-selective Rab-binding assay, and in vitro GAP activity assay","pmids":["17099728"],"confidence":"High","gaps":["Directly contradicts the 2007 finding of no GAP activity in vitro","Cellular substrate specificity not confirmed"]},{"year":2007,"claim":"Characterizing GTP-dependent Rab11 binding and competition with the effector FIP3 defined EVI5 as a Rab11 partner regulating cytokinetic recycling, while questioning whether it acts catalytically.","evidence":"Y2H, Co-IP, Biacore SPR, in vitro GAP assay (negative), RNAi with confocal imaging of FIP3 mislocalization","pmids":["17229837"],"confidence":"High","gaps":["No GAP activity detected, unreconciled with the 2006 Oncogene result","Mechanism of FIP3 competition not structurally defined"]},{"year":2009,"claim":"Showing EVI5 overexpression maintains Runx1-deficient hematopoietic stem cells linked it to leukemogenesis as a cooperating oncogenic event in vivo.","evidence":"retroviral insertional mutagenesis screen with conditional Runx1 knockout mice and retroviral EVI5 rescue","pmids":["20008790"],"confidence":"Medium","gaps":["Molecular mechanism connecting EVI5 to stem-cell maintenance not defined","Whether GAP or Emi1-stabilizing activity underlies the effect unknown"]},{"year":2012,"claim":"Genetic dissection in Drosophila border cells confirmed that EVI5 Rab-GAP activity directs Rab11-dependent polarization of guidance receptors, establishing an in vivo role in directed cell migration.","evidence":"Rab-GAP genetic screen, loss- and gain-of-function genetics with TBC-domain requirement, in vivo imaging and Rab11 activity reporters","pmids":["22778279"],"confidence":"High","gaps":["Conservation of the migration role in mammalian cells not shown","Identity of the specific guidance receptors trafficked not fully resolved"]},{"year":2015,"claim":"Mapping how a multiple sclerosis risk SNP remodels the EVI5 coiled-coil interactome implicated its scaffolding domain in disease and revealed a risk-variant-specific interaction with the lipid enzyme SGPL1.","evidence":"fine-mapping/meta-analysis, IP-MS comparison of wild-type vs risk variant, and coiled-coil structural modeling","pmids":["26433934"],"confidence":"Medium","gaps":["Functional consequence of the SGPL1 interaction for MS pathogenesis not established","Link between altered interactome and EVI5's GAP/cell-cycle roles unclear"]},{"year":2020,"claim":"Identifying EVI5 as an antagonist of FBXW7-mediated c-Myc degradation extended its proteostatic role from Emi1 to a major oncoprotein, providing a mechanism for its tumor-promoting activity.","evidence":"Co-IP, CRISPR knockout, luciferase reporter, flow cytometry, and xenograft in laryngeal squamous cell carcinoma","pmids":["32047362"],"confidence":"Medium","gaps":["Whether c-Myc stabilization requires the TBC domain or coiled-coil region not determined","Direct vs indirect antagonism of FBXW7 not biochemically resolved"]},{"year":2020,"claim":"Co-IP of EVI5 with TGF-beta receptors and Emi1 in NSCLC and its placement downstream of miR-486-5p extended EVI5 into TGF-beta/Smad-driven migration, though the interactions rest on single binding assays.","evidence":"Co-IP, siRNA knockdown, wound-healing/transwell invasion assays, and xenograft","pmids":["32393392"],"confidence":"Low","gaps":["Single Co-IP without reciprocal or mutagenesis validation of the TGF-betaR interaction","Mechanistic link between EVI5 and Smad signaling not dissected"]},{"year":2022,"claim":"Demonstrating a requirement for Evi5 in Xenopus limb and tail regeneration tied it to blastema proliferation through PDGF/TGF-beta signaling and lysine demethylases, broadening its role into developmental regeneration.","evidence":"morpholino knockdown in tadpoles, RNA-seq, proliferation/apoptosis assays, and Kdm6b/Kdm7a knockdown epistasis","pmids":["36605717"],"confidence":"Medium","gaps":["Whether regeneration role depends on GAP activity not tested","Direct targets connecting EVI5 to demethylase expression unknown"]},{"year":2024,"claim":"Linking Evi5-dependent endosome recycling to transferrin trafficking and iron/heme homeostasis, including a physical interaction with ferritin, connected its trafficking function to organismal metabolism.","evidence":"RNAi depletion in Drosophila prothoracic gland, EM of vesicle morphology, ferritin Co-IP, and ferritin injection rescue","pmids":["38744835"],"confidence":"Medium","gaps":["Whether iron trafficking depends on Rab11-GAP activity not established","Mammalian conservation of the ferritin interaction not shown"]},{"year":2025,"claim":"Recent oncology studies place EVI5 in EVI5–Rab11–PD-L1 and ERK1/2–c-Myc axes in lung adenocarcinoma, but rely on single Co-IPs and inhibitor epistasis without direct mechanistic dissection.","evidence":"Co-IP, CRISPR knockout, overexpression, ERK1/2 inhibitor rescue, Western blot, qRT-PCR","pmids":["41258404","40525449"],"confidence":"Low","gaps":["No mechanism connecting Rab11 to PD-L1 expression","No direct biochemical interaction between EVI5 and ERK1/2 demonstrated"]},{"year":null,"claim":"It remains unresolved whether EVI5 acts as a catalytic Rab11-GAP or solely as a GTP-dependent Rab11-binding/effector-competing scaffold, and how its trafficking, Emi1/c-Myc stabilizing, and centrosomal functions are mechanistically unified within one protein.","evidence":"no single study reconciles the contradictory in vitro GAP results or integrates the trafficking and proteostatic activities","pmids":[],"confidence":"Low","gaps":["Contradiction between positive and negative in vitro GAP activity unresolved","No structural model of full-length EVI5 with its partners","Domain requirements for each distinct function not systematically mapped"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[2,5]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,2,5]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[3]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[4]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[1,11]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,4]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[1,5]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,8]}],"complexes":["chromosomal passenger complex"],"partners":["RAB11A","RAB11B","FBXO5","MYC","INCENP","AURKB","BIRC5","FBXW7"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O60447","full_name":"Ecotropic viral integration site 5 protein homolog","aliases":["Neuroblastoma stage 4S gene protein"],"length_aa":810,"mass_kda":92.9,"function":"Functions as a regulator of cell cycle progression by stabilizing the FBXO5 protein and promoting cyclin-A accumulation during interphase. May play a role in cytokinesis","subcellular_location":"Nucleus; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Cytoplasm, cytoskeleton, spindle","url":"https://www.uniprot.org/uniprotkb/O60447/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/EVI5","classification":"Not Classified","n_dependent_lines":9,"n_total_lines":1208,"dependency_fraction":0.0074503311258278145},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"RAB11A","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/EVI5","total_profiled":1310},"omim":[{"mim_id":"606013","title":"F-BOX ONLY PROTEIN 5; FBXO5","url":"https://www.omim.org/entry/606013"},{"mim_id":"602942","title":"ECOTROPIC VIRAL INTEGRATION SITE 5; EVI5","url":"https://www.omim.org/entry/602942"},{"mim_id":"256700","title":"NEUROBLASTOMA, SUSCEPTIBILITY TO, 1; NBLST1","url":"https://www.omim.org/entry/256700"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Golgi apparatus","reliability":"Approved"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/EVI5"},"hgnc":{"alias_symbol":["NB4S"],"prev_symbol":[]},"alphafold":{"accession":"O60447","domains":[{"cath_id":"1.10.8.270","chopping":"183-283","consensus_level":"medium","plddt":94.4435,"start":183,"end":283},{"cath_id":"-","chopping":"424-451_458-498","consensus_level":"medium","plddt":84.0884,"start":424,"end":498},{"cath_id":"-","chopping":"504-545_560-687","consensus_level":"medium","plddt":87.8709,"start":504,"end":687}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O60447","model_url":"https://alphafold.ebi.ac.uk/files/AF-O60447-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O60447-F1-predicted_aligned_error_v6.png","plddt_mean":74.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=EVI5","jax_strain_url":"https://www.jax.org/strain/search?query=EVI5"},"sequence":{"accession":"O60447","fasta_url":"https://rest.uniprot.org/uniprotkb/O60447.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O60447/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O60447"}},"corpus_meta":[{"pmid":"20008790","id":"PMC_20008790","title":"Stem cell exhaustion due to Runx1 deficiency is prevented by Evi5 activation in leukemogenesis.","date":"2009","source":"Blood","url":"https://pubmed.ncbi.nlm.nih.gov/20008790","citation_count":84,"is_preprint":false},{"pmid":"16439210","id":"PMC_16439210","title":"The evi5 oncogene regulates cyclin accumulation by stabilizing the anaphase-promoting complex inhibitor emi1.","date":"2006","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/16439210","citation_count":83,"is_preprint":false},{"pmid":"25537516","id":"PMC_25537516","title":"MicroRNA-135b, a HSF1 target, promotes tumor invasion and metastasis by regulating RECK and EVI5 in hepatocellular carcinoma.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/25537516","citation_count":67,"is_preprint":false},{"pmid":"18401352","id":"PMC_18401352","title":"EVI5 is a risk gene for multiple sclerosis.","date":"2008","source":"Genes and immunity","url":"https://pubmed.ncbi.nlm.nih.gov/18401352","citation_count":62,"is_preprint":false},{"pmid":"17229837","id":"PMC_17229837","title":"Identification of Rab11 as a small GTPase binding protein for the Evi5 oncogene.","date":"2007","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/17229837","citation_count":61,"is_preprint":false},{"pmid":"22778279","id":"PMC_22778279","title":"Evi5 promotes collective cell migration through its Rab-GAP activity.","date":"2012","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/22778279","citation_count":54,"is_preprint":false},{"pmid":"17099728","id":"PMC_17099728","title":"The EVI5 TBC domain provides the GTPase-activating protein motif for RAB11.","date":"2006","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/17099728","citation_count":50,"is_preprint":false},{"pmid":"9618176","id":"PMC_9618176","title":"NB4S, a member of the TBC1 domain family of genes, is truncated as a result of a constitutional t(1;10)(p22;q21) chromosome translocation in a patient with stage 4S neuroblastoma.","date":"1998","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/9618176","citation_count":48,"is_preprint":false},{"pmid":"7474133","id":"PMC_7474133","title":"Evi-5, a common site of retroviral integration in AKXD T-cell lymphomas, maps near Gfi-1 on mouse chromosome 5.","date":"1995","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/7474133","citation_count":44,"is_preprint":false},{"pmid":"23669355","id":"PMC_23669355","title":"The Evi5 family in cellular physiology and pathology.","date":"2013","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/23669355","citation_count":36,"is_preprint":false},{"pmid":"20087403","id":"PMC_20087403","title":"Tag-SNP analysis of the GFI1-EVI5-RPL5-FAM69 risk locus for multiple sclerosis.","date":"2010","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/20087403","citation_count":24,"is_preprint":false},{"pmid":"16033705","id":"PMC_16033705","title":"EVI5 is a novel centrosomal protein that binds to alpha- and gamma-tubulin.","date":"2005","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/16033705","citation_count":23,"is_preprint":false},{"pmid":"9070650","id":"PMC_9070650","title":"Proviral integrations at the Evi5 locus disrupt a novel 90 kDa protein with homology to the Tre2 oncogene and cell-cycle regulatory proteins.","date":"1997","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/9070650","citation_count":22,"is_preprint":false},{"pmid":"16764853","id":"PMC_16764853","title":"EVI5 protein associates with the INCENP-aurora B kinase-survivin chromosomal passenger complex and is involved in the completion of cytokinesis.","date":"2006","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/16764853","citation_count":19,"is_preprint":false},{"pmid":"32393392","id":"PMC_32393392","title":"EVI5 is an oncogene that regulates the proliferation and metastasis of NSCLC cells.","date":"2020","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/32393392","citation_count":16,"is_preprint":false},{"pmid":"19282512","id":"PMC_19282512","title":"The NBPF1 promoter has been recruited from the unrelated EVI5 gene before simian radiation.","date":"2009","source":"Molecular biology and evolution","url":"https://pubmed.ncbi.nlm.nih.gov/19282512","citation_count":14,"is_preprint":false},{"pmid":"26433934","id":"PMC_26433934","title":"A non-synonymous single-nucleotide polymorphism associated with multiple sclerosis risk affects the EVI5 interactome.","date":"2015","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26433934","citation_count":14,"is_preprint":false},{"pmid":"23910958","id":"PMC_23910958","title":"GFI1B, EVI5, MYB--additional genes that cooperate with the human BCL6 gene to promote the development of lymphomas.","date":"2013","source":"Blood cells, molecules & diseases","url":"https://pubmed.ncbi.nlm.nih.gov/23910958","citation_count":9,"is_preprint":false},{"pmid":"38744835","id":"PMC_38744835","title":"Drosophila Evi5 is a critical regulator of intracellular iron transport via transferrin and ferritin interactions.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/38744835","citation_count":8,"is_preprint":false},{"pmid":"32047362","id":"PMC_32047362","title":"The Evi5 oncogene promotes laryngeal cancer cells proliferation by stabilizing c-Myc protein.","date":"2020","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/32047362","citation_count":8,"is_preprint":false},{"pmid":"29141798","id":"PMC_29141798","title":"Ecotropic Viral Integration Site 5 (EVI5) variants are associated with multiple sclerosis in Iranian population.","date":"2017","source":"Multiple sclerosis and related disorders","url":"https://pubmed.ncbi.nlm.nih.gov/29141798","citation_count":6,"is_preprint":false},{"pmid":"29036808","id":"PMC_29036808","title":"Ecotropic Viral Integration Site 5 (EVI5) expression analysis in multiple sclerosis patients.","date":"2017","source":"Human antibodies","url":"https://pubmed.ncbi.nlm.nih.gov/29036808","citation_count":4,"is_preprint":false},{"pmid":"25800843","id":"PMC_25800843","title":"In vitro and in vivo characterization of the Rab11-GAP activity of Drosophila Evi5.","date":"2015","source":"Methods in molecular biology (Clifton, N.J.)","url":"https://pubmed.ncbi.nlm.nih.gov/25800843","citation_count":4,"is_preprint":false},{"pmid":"39940949","id":"PMC_39940949","title":"Novel EVI5::BRAF Gene Fusion in Infantile Fibrosarcoma: A Case Report and Review of Literature.","date":"2025","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/39940949","citation_count":2,"is_preprint":false},{"pmid":"36605717","id":"PMC_36605717","title":"Evi5 is required for Xenopus limb and tail regeneration.","date":"2022","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/36605717","citation_count":2,"is_preprint":false},{"pmid":"41258404","id":"PMC_41258404","title":"EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/41258404","citation_count":0,"is_preprint":false},{"pmid":"40525449","id":"PMC_40525449","title":"EVI5 Drives Lung Adenocarcinoma Progression through Modulation of the ERK1/2-c-Myc Signaling Axis.","date":"2025","source":"Current cancer drug targets","url":"https://pubmed.ncbi.nlm.nih.gov/40525449","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13797,"output_tokens":4101,"usd":0.051453,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11933,"output_tokens":4955,"usd":0.09177,"stage2_stop_reason":"end_turn"},"total_usd":0.143223,"stage1_batch_id":"msgbatch_01AsR7g8dGTEiEw9U83bJeWn","stage2_batch_id":"msgbatch_01YV5ZyxtFNUZEjxRcpd9jzb","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2006,\n      \"finding\": \"Evi5 stabilizes Emi1 (an APC/C inhibitor) by binding adjacent to Emi1's DSGxxS degron, blocking both degron phosphorylation by Polo-like kinases and subsequent betaTrCP binding, thereby antagonizing SCF(betaTrCP)-dependent Emi1 ubiquitination and destruction. Evi5 protein accumulates in early G1 following Plk1 destruction and is itself degraded in a Plk1- and ubiquitin-dependent manner in early mitosis. Ablation of Evi5 causes precocious Emi1 degradation, premature APC/C activation, cyclin destruction, centrosome overduplication, and mitotic catastrophe.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, cell-cycle imaging, kinase assays\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal biochemical methods (binding assay, ubiquitination assay, mutagenesis of degron, kinase assays) plus cellular phenotypic rescue in a single rigorous study\",\n      \"pmids\": [\"16439210\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Evi5 binds Rab11a and Rab11b in a GTP-dependent manner (identified by yeast two-hybrid, co-immunoprecipitation, and Biacore surface plasmon resonance), but displays no GTPase-activating activity toward Rab11 in vitro. Evi5 colocalizes with Rab11 in vivo; Rab11 effector proteins (e.g., FIP3) compete with Evi5 for Rab11 binding. Ablation of Evi5 by RNAi causes mislocalization of FIP3 at the abscission site during cytokinesis.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, Biacore SPR, in vitro GAP assay, RNAi knockdown, confocal microscopy\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal binding methods (Y2H, Co-IP, SPR), in vitro GAP assay, and cellular phenotype with RNAi in a single study\",\n      \"pmids\": [\"17229837\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EVI5's TBC domain functions as a GTPase-activating protein (GAP) for Rab11: EVI5 preferentially binds the GTP-bound form of Rab11 and stimulates Rab11 GTPase activity in a GAP activity assay, as demonstrated by mass spectrometry-identified complex formation and a specific Rab-binding/GAP assay.\",\n      \"method\": \"Linear ion trap mass spectrometry (protein complex identification), GTP-form-selective Rab-binding assay, in vitro GAP activity assay\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro GAP assay plus MS-based complex identification, single lab but two orthogonal methods; note this finding contradicts Westlake et al. 2007 which found no GAP activity, but this paper reports positive GAP stimulation\",\n      \"pmids\": [\"17099728\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"EVI5 localizes to pericentriolar material (centrosome) in interphase cells and exists in protein complexes with both alpha- and gamma-tubulin, with both interactions mapping to the N-terminal region of EVI5. EVI5 can form dimers as shown by FPLC analysis.\",\n      \"method\": \"Confocal immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, FPLC gel filtration\",\n      \"journal\": \"Genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal Co-IP and GST pull-down plus localization, single lab\",\n      \"pmids\": [\"16033705\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"EVI5 has a dynamic mitotic distribution: it associates with the mitotic spindle through anaphase and localizes to the midzone and midbody until completion of cytokinesis. EVI5 undergoes phosphorylation in early mitosis and proteolytic cleavage during late mitosis/cytokinesis. EVI5 co-immunoprecipitates and co-purifies (GST pull-down) with the chromosomal passenger complex (INCENP, Aurora B kinase, survivin). siRNA knockdown of EVI5 produces a multinucleate phenotype, establishing an essential role in cytokinesis completion.\",\n      \"method\": \"Immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, siRNA knockdown, Western blot for phosphorylation/cleavage\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal Co-IP, GST pull-down, and siRNA phenotype, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"16764853\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Drosophila Evi5 promotes collective border cell migration through its Rab-GAP activity directed at Rab11. Both loss and gain of Evi5 function block border cell migration by disrupting Rab11-dependent polarization of active guidance receptors. The TBC/Rab-GAP domain is required for this function.\",\n      \"method\": \"Drosophila genetic screen for Rab-GAP proteins, loss-of-function and gain-of-function genetics, in vivo imaging of border cell migration, Rab11 activity reporters\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic epistasis with domain-requirement validation and in vivo imaging in Drosophila ortholog, replicates the Rab11-GAP function\",\n      \"pmids\": [\"22778279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"The human EVI5 gene (NB4S) encodes an 810 amino acid protein with a TBC1 box motif (homologous to genes involved in cell growth/differentiation) and C-terminal coiled-coil domains. It is disrupted by a constitutional t(1;10)(p22;q21) translocation in a neuroblastoma patient, generating a fusion transcript that combines the TBC1 motif of NB4S with a polyadenylation signal from TRNG10, potentially producing a truncated oncogenic protein.\",\n      \"method\": \"Molecular cloning of translocation breakpoints, sequence analysis, Northern blot expression analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — direct molecular cloning and sequencing of the translocation breakpoint, single lab\",\n      \"pmids\": [\"9618176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"EVI5 overexpression prevents Runx1-deficient hematopoietic stem cell (HSC) exhaustion in mice, identified by retroviral insertional mutagenesis as a cooperating genetic alteration in leukemia development. EVI5 activation maintains pre-leukemic stem cell populations in the context of Runx1 loss.\",\n      \"method\": \"Retroviral insertional mutagenesis screen, conditional Runx1 knockout mouse model, retroviral EVI5 overexpression rescue experiment\",\n      \"journal\": \"Blood\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with in vivo rescue experiment, single lab\",\n      \"pmids\": [\"20008790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Evi5 interacts with c-Myc and antagonizes FBXW7 E3 ligase-mediated ubiquitination and degradation of c-Myc protein, thereby stabilizing c-Myc and promoting its transcriptional activity in laryngeal squamous cell carcinoma cells. CRISPR-mediated Evi5 knockout decreases c-Myc levels, causes G1 arrest, and reduces tumor growth in vivo.\",\n      \"method\": \"Co-immunoprecipitation, CRISPR knockout, luciferase transcriptional reporter assay, flow cytometry, xenograft mouse model\",\n      \"journal\": \"Cancer cell international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — Co-IP for interaction, CRISPR KO with multiple phenotypic readouts, single lab\",\n      \"pmids\": [\"32047362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"EVI5 interacts with TGF-β receptor I and TGF-β receptor II (shown by co-immunoprecipitation), and the miR-486-5p–EVI5 axis affects NSCLC migration and invasion through the TGF-β/Smad signaling pathway. EVI5 also interacts with Emi1 in NSCLC cells.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, wound healing/transwell invasion assays, xenograft mouse model\",\n      \"journal\": \"Journal of experimental & clinical cancer research : CR\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP for TGFβR interaction, single lab, no mutagenesis or reconstitution to confirm mechanistic link\",\n      \"pmids\": [\"32393392\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"An exonic SNP in EVI5 associated with multiple sclerosis risk induces changes in superficial hydrophobicity patterns of the EVI5 coiled-coil domain, altering the EVI5 interactome. Immunoprecipitation followed by mass spectrometry of wild-type vs. risk-variant EVI5 revealed disease-specific interactors linked to lipid metabolism, including a novel interaction with sphingosine 1-phosphate lyase (SGPL1) exclusive to the risk variant.\",\n      \"method\": \"Fine mapping/meta-analysis, co-immunoprecipitation followed by mass spectrometry, structural modeling of coiled-coil domain\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — IP-MS interactome comparison with functional variant, single lab, novel interaction identified by two orthogonal approaches (IP-MS and structural analysis)\",\n      \"pmids\": [\"26433934\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Drosophila Evi5 is required for intracellular iron trafficking: Evi5 depletion in the prothoracic gland disrupts vesicle morphology, blocks endosome recycling, and impairs transferrin-1 trafficking, reducing cellular iron and impairing heme synthesis. Evi5 physically interacts with ferritin, and ferritin injection rescues developmental delays caused by Evi5 depletion.\",\n      \"method\": \"RNAi depletion in Drosophila prothoracic gland, electron microscopy of vesicle morphology, ferritin co-immunoprecipitation/physical interaction assay, ferritin injection rescue experiment\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (RNAi, EM, physical interaction, rescue), single lab, Drosophila ortholog\",\n      \"pmids\": [\"38744835\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVI5 interacts with Rab11 and upregulates PD-L1 expression in lung adenocarcinoma cells. EVI5 knockout suppresses Rab11 and PD-L1 expression, while EVI5 overexpression promotes their expression, defining an EVI5–Rab11–PD-L1 regulatory axis.\",\n      \"method\": \"Co-immunoprecipitation, CRISPR knockout, overexpression, Western blot\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP for interaction, single lab, no mechanistic dissection of how Rab11 connects to PD-L1\",\n      \"pmids\": [\"41258404\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"EVI5 promotes lung adenocarcinoma progression through activation of the ERK1/2–c-Myc signaling axis; EVI5 knockout reduces ERK1/2 and c-Myc activity, and an ERK1/2 pathway inhibitor phenocopies EVI5 loss, validated by functional rescue experiments.\",\n      \"method\": \"CRISPR knockout, overexpression, Western blot, ERK1/2 inhibitor functional rescue, immunofluorescence, qRT-PCR\",\n      \"journal\": \"Current cancer drug targets\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — genetic epistasis via inhibitor rescue, single lab, no direct biochemical interaction between EVI5 and ERK1/2 demonstrated\",\n      \"pmids\": [\"40525449\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Evi5 is required for Xenopus limb and tail regeneration: morpholino-mediated Evi5 knockdown reduces blastema cell proliferation, causes apoptosis, and blocks blastema formation. RNA-seq reveals Evi5 knockdown downregulates PDGFα and TGFβ signaling pathways as well as lysine demethylases Kdm6b and Kdm7a, which are themselves required for limb regeneration.\",\n      \"method\": \"Morpholino antisense knockdown in Xenopus tadpoles, RNA-sequencing, proliferation and apoptosis assays, Kdm6b/Kdm7a knockdown epistasis\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — morpholino knockdown with RNA-seq and epistasis experiments in Xenopus ortholog, single lab\",\n      \"pmids\": [\"36605717\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"EVI5 is a TBC-domain-containing protein that functions as a Rab11 GTPase-activating protein (or GTP-dependent Rab11 binding partner) to regulate endosomal recycling, cytokinesis, and cell migration; it also acts as a cell cycle regulator by stabilizing Emi1 (blocking SCF(betaTrCP)-mediated ubiquitination) to control APC/C activity and cyclin accumulation, associates with the Aurora B/INCENP/survivin chromosomal passenger complex at the midbody, localizes to centrosomes via interactions with alpha- and gamma-tubulin, and can stabilize oncoproteins such as c-Myc by antagonizing FBXW7-dependent degradation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"EVI5 is a TBC-domain protein that integrates membrane-trafficking control with cell-cycle regulation, acting at the interface of Rab11-dependent endosomal recycling and the mitotic machinery [#1, #0]. Through its TBC/Rab-GAP domain EVI5 binds Rab11 selectively in its GTP-bound state and governs Rab11-dependent recycling, polarizing guidance-receptor delivery during collective cell migration [#1, #5]; whether it stimulates Rab11 GTP hydrolysis is reported divergently, with one study finding GAP activity [#2] and another finding GTP-dependent binding without measurable GAP activity in vitro [#1]. In parallel, EVI5 controls cell-cycle progression by binding adjacent to the DSGxxS degron of the APC/C inhibitor Emi1, blocking Plk1-mediated degron phosphorylation and subsequent betaTrCP recruitment, thereby stabilizing Emi1 and restraining premature APC/C activation; EVI5 itself accumulates in early G1 and is degraded in a Plk1- and ubiquitin-dependent manner in early mitosis, and its loss causes precocious Emi1 destruction, cyclin loss, centrosome overduplication, and mitotic catastrophe [#0]. EVI5 has a dynamic mitotic distribution, localizing to centrosomal pericentriolar material via N-terminal interactions with alpha- and gamma-tubulin and tracking the spindle, midzone, and midbody, where it associates with the chromosomal passenger complex (INCENP, Aurora B, survivin) and is required for cytokinesis completion [#3, #4]. EVI5 was first identified as a gene (NB4S) disrupted by a constitutional translocation in neuroblastoma [#6], and subsequent work links it to oncogenic and developmental roles: it antagonizes FBXW7-mediated ubiquitination of c-Myc to stabilize the oncoprotein [#8], maintains pre-leukemic hematopoietic stem cells in the setting of Runx1 loss [#7], and is required for vertebrate appendage regeneration through PDGF/TGF-beta signaling and lysine demethylases [#14].\",\n  \"teleology\": [\n    {\n      \"year\": 1998,\n      \"claim\": \"Establishing the molecular identity of EVI5 — that the gene encodes a TBC1-motif, coiled-coil protein disrupted in cancer — provided the first structural framework for its later-defined GAP and scaffolding functions.\",\n      \"evidence\": \"molecular cloning and sequencing of a t(1;10) translocation breakpoint in a neuroblastoma patient, with Northern expression analysis\",\n      \"pmids\": [\"9618176\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Did not establish the biochemical activity of the TBC1 motif\", \"Oncogenic consequence of the fusion transcript inferred but not functionally tested\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Localizing EVI5 to the centrosome and showing it binds alpha- and gamma-tubulin defined a structural/scaffolding role at the pericentriolar material before its enzymatic activity was known.\",\n      \"evidence\": \"confocal immunofluorescence, reciprocal Co-IP, GST pull-down, and FPLC gel filtration\",\n      \"pmids\": [\"16033705\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of tubulin binding not tested\", \"Single lab, no in vivo validation of centrosomal function\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Identifying EVI5 as a stabilizer of the APC/C inhibitor Emi1 placed it directly in cell-cycle control, explaining why its loss triggers premature APC/C activation and mitotic catastrophe.\",\n      \"evidence\": \"Co-IP, in vitro ubiquitination assay, degron mutagenesis, kinase assays, siRNA knockdown with cell-cycle imaging\",\n      \"pmids\": [\"16439210\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of degron-adjacent binding not resolved\", \"Relationship between Emi1-stabilizing and Rab-GAP functions of the same protein not integrated\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Tracking EVI5's mitotic distribution and its association with the chromosomal passenger complex established a midbody role essential for cytokinesis completion, complementing its earlier centrosomal localization.\",\n      \"evidence\": \"immunofluorescence across mitosis, Co-IP/GST pull-down with INCENP/Aurora B/survivin, siRNA producing a multinucleate phenotype, Western blots for phosphorylation/cleavage\",\n      \"pmids\": [\"16764853\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional significance of CPC association not dissected\", \"Identity of the protease cleaving EVI5 in late mitosis unknown\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Demonstrating that the EVI5 TBC domain stimulates Rab11 GTP hydrolysis assigned it a Rab11-GAP enzymatic activity, linking it to endosomal trafficking control.\",\n      \"evidence\": \"linear ion-trap MS complex identification, GTP-form-selective Rab-binding assay, and in vitro GAP activity assay\",\n      \"pmids\": [\"17099728\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Directly contradicts the 2007 finding of no GAP activity in vitro\", \"Cellular substrate specificity not confirmed\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Characterizing GTP-dependent Rab11 binding and competition with the effector FIP3 defined EVI5 as a Rab11 partner regulating cytokinetic recycling, while questioning whether it acts catalytically.\",\n      \"evidence\": \"Y2H, Co-IP, Biacore SPR, in vitro GAP assay (negative), RNAi with confocal imaging of FIP3 mislocalization\",\n      \"pmids\": [\"17229837\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No GAP activity detected, unreconciled with the 2006 Oncogene result\", \"Mechanism of FIP3 competition not structurally defined\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Showing EVI5 overexpression maintains Runx1-deficient hematopoietic stem cells linked it to leukemogenesis as a cooperating oncogenic event in vivo.\",\n      \"evidence\": \"retroviral insertional mutagenesis screen with conditional Runx1 knockout mice and retroviral EVI5 rescue\",\n      \"pmids\": [\"20008790\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism connecting EVI5 to stem-cell maintenance not defined\", \"Whether GAP or Emi1-stabilizing activity underlies the effect unknown\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Genetic dissection in Drosophila border cells confirmed that EVI5 Rab-GAP activity directs Rab11-dependent polarization of guidance receptors, establishing an in vivo role in directed cell migration.\",\n      \"evidence\": \"Rab-GAP genetic screen, loss- and gain-of-function genetics with TBC-domain requirement, in vivo imaging and Rab11 activity reporters\",\n      \"pmids\": [\"22778279\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Conservation of the migration role in mammalian cells not shown\", \"Identity of the specific guidance receptors trafficked not fully resolved\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Mapping how a multiple sclerosis risk SNP remodels the EVI5 coiled-coil interactome implicated its scaffolding domain in disease and revealed a risk-variant-specific interaction with the lipid enzyme SGPL1.\",\n      \"evidence\": \"fine-mapping/meta-analysis, IP-MS comparison of wild-type vs risk variant, and coiled-coil structural modeling\",\n      \"pmids\": [\"26433934\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the SGPL1 interaction for MS pathogenesis not established\", \"Link between altered interactome and EVI5's GAP/cell-cycle roles unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identifying EVI5 as an antagonist of FBXW7-mediated c-Myc degradation extended its proteostatic role from Emi1 to a major oncoprotein, providing a mechanism for its tumor-promoting activity.\",\n      \"evidence\": \"Co-IP, CRISPR knockout, luciferase reporter, flow cytometry, and xenograft in laryngeal squamous cell carcinoma\",\n      \"pmids\": [\"32047362\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether c-Myc stabilization requires the TBC domain or coiled-coil region not determined\", \"Direct vs indirect antagonism of FBXW7 not biochemically resolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Co-IP of EVI5 with TGF-beta receptors and Emi1 in NSCLC and its placement downstream of miR-486-5p extended EVI5 into TGF-beta/Smad-driven migration, though the interactions rest on single binding assays.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, wound-healing/transwell invasion assays, and xenograft\",\n      \"pmids\": [\"32393392\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single Co-IP without reciprocal or mutagenesis validation of the TGF-betaR interaction\", \"Mechanistic link between EVI5 and Smad signaling not dissected\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating a requirement for Evi5 in Xenopus limb and tail regeneration tied it to blastema proliferation through PDGF/TGF-beta signaling and lysine demethylases, broadening its role into developmental regeneration.\",\n      \"evidence\": \"morpholino knockdown in tadpoles, RNA-seq, proliferation/apoptosis assays, and Kdm6b/Kdm7a knockdown epistasis\",\n      \"pmids\": [\"36605717\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether regeneration role depends on GAP activity not tested\", \"Direct targets connecting EVI5 to demethylase expression unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linking Evi5-dependent endosome recycling to transferrin trafficking and iron/heme homeostasis, including a physical interaction with ferritin, connected its trafficking function to organismal metabolism.\",\n      \"evidence\": \"RNAi depletion in Drosophila prothoracic gland, EM of vesicle morphology, ferritin Co-IP, and ferritin injection rescue\",\n      \"pmids\": [\"38744835\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether iron trafficking depends on Rab11-GAP activity not established\", \"Mammalian conservation of the ferritin interaction not shown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Recent oncology studies place EVI5 in EVI5–Rab11–PD-L1 and ERK1/2–c-Myc axes in lung adenocarcinoma, but rely on single Co-IPs and inhibitor epistasis without direct mechanistic dissection.\",\n      \"evidence\": \"Co-IP, CRISPR knockout, overexpression, ERK1/2 inhibitor rescue, Western blot, qRT-PCR\",\n      \"pmids\": [\"41258404\", \"40525449\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No mechanism connecting Rab11 to PD-L1 expression\", \"No direct biochemical interaction between EVI5 and ERK1/2 demonstrated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved whether EVI5 acts as a catalytic Rab11-GAP or solely as a GTP-dependent Rab11-binding/effector-competing scaffold, and how its trafficking, Emi1/c-Myc stabilizing, and centrosomal functions are mechanistically unified within one protein.\",\n      \"evidence\": \"no single study reconciles the contradictory in vitro GAP results or integrates the trafficking and proteostatic activities\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Contradiction between positive and negative in vitro GAP activity unresolved\", \"No structural model of full-length EVI5 with its partners\", \"Domain requirements for each distinct function not systematically mapped\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [2, 5]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 2, 5]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [1, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [1, 5]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 8]}\n    ],\n    \"complexes\": [\"chromosomal passenger complex\"],\n    \"partners\": [\"RAB11A\", \"RAB11B\", \"FBXO5\", \"MYC\", \"INCENP\", \"AURKB\", \"BIRC5\", \"FBXW7\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}