Affinage

EVI5

Ecotropic viral integration site 5 protein homolog · UniProt O60447

Length
810 aa
Mass
92.9 kDa
Annotated
2026-06-09
27 papers in source corpus 15 papers cited in narrative 15 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 5/5 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

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).

Mechanistic history

Synthesis pass · year-by-year structured walk · 14 steps
  1. 1998 Medium

    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

    PMID:9618176

    Open questions at the time
    • Did not establish the biochemical activity of the TBC1 motif
    • Oncogenic consequence of the fusion transcript inferred but not functionally tested
  2. 2005 Medium

    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

    PMID:16033705

    Open questions at the time
    • Functional consequence of tubulin binding not tested
    • Single lab, no in vivo validation of centrosomal function
  3. 2006 High

    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

    PMID:16439210

    Open questions at the time
    • Structural basis of degron-adjacent binding not resolved
    • Relationship between Emi1-stabilizing and Rab-GAP functions of the same protein not integrated
  4. 2006 Medium

    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

    PMID:16764853

    Open questions at the time
    • Functional significance of CPC association not dissected
    • Identity of the protease cleaving EVI5 in late mitosis unknown
  5. 2006 High

    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

    PMID:17099728

    Open questions at the time
    • Directly contradicts the 2007 finding of no GAP activity in vitro
    • Cellular substrate specificity not confirmed
  6. 2007 High

    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

    PMID:17229837

    Open questions at the time
    • No GAP activity detected, unreconciled with the 2006 Oncogene result
    • Mechanism of FIP3 competition not structurally defined
  7. 2009 Medium

    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

    PMID:20008790

    Open questions at the time
    • Molecular mechanism connecting EVI5 to stem-cell maintenance not defined
    • Whether GAP or Emi1-stabilizing activity underlies the effect unknown
  8. 2012 High

    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

    PMID:22778279

    Open questions at the time
    • Conservation of the migration role in mammalian cells not shown
    • Identity of the specific guidance receptors trafficked not fully resolved
  9. 2015 Medium

    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

    PMID:26433934

    Open questions at the time
    • Functional consequence of the SGPL1 interaction for MS pathogenesis not established
    • Link between altered interactome and EVI5's GAP/cell-cycle roles unclear
  10. 2020 Medium

    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

    PMID:32047362

    Open questions at the time
    • Whether c-Myc stabilization requires the TBC domain or coiled-coil region not determined
    • Direct vs indirect antagonism of FBXW7 not biochemically resolved
  11. 2020 Low

    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

    PMID:32393392

    Open questions at the time
    • Single Co-IP without reciprocal or mutagenesis validation of the TGF-betaR interaction
    • Mechanistic link between EVI5 and Smad signaling not dissected
  12. 2022 Medium

    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

    PMID:36605717

    Open questions at the time
    • Whether regeneration role depends on GAP activity not tested
    • Direct targets connecting EVI5 to demethylase expression unknown
  13. 2024 Medium

    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

    PMID:38744835

    Open questions at the time
    • Whether iron trafficking depends on Rab11-GAP activity not established
    • Mammalian conservation of the ferritin interaction not shown
  14. 2025 Low

    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

    PMID:40525449 PMID:41258404

    Open questions at the time
    • No mechanism connecting Rab11 to PD-L1 expression
    • No direct biochemical interaction between EVI5 and ERK1/2 demonstrated

Open questions

Synthesis pass · forward-looking unresolved questions
  • 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.
  • 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

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0098772 molecular function regulator activity 3 GO:0003924 GTPase activity 2 GO:0060090 molecular adaptor activity 2 GO:0008092 cytoskeletal protein binding 1
Localization
GO:0005768 endosome 2 GO:0005815 microtubule organizing center 1 GO:0005856 cytoskeleton 1
Pathway
R-HSA-1640170 Cell Cycle 2 R-HSA-392499 Metabolism of proteins 2 R-HSA-5653656 Vesicle-mediated transport 2
Complex memberships
chromosomal passenger complex

Evidence

Reading pass · 15 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2006 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. Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, cell-cycle imaging, kinase assays Cell High 16439210
2007 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. Yeast two-hybrid, co-immunoprecipitation, Biacore SPR, in vitro GAP assay, RNAi knockdown, confocal microscopy Proceedings of the National Academy of Sciences of the United States of America High 17229837
2006 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. Linear ion trap mass spectrometry (protein complex identification), GTP-form-selective Rab-binding assay, in vitro GAP activity assay Oncogene High 17099728
2005 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. Confocal immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, FPLC gel filtration Genomics Medium 16033705
2006 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. Immunofluorescence microscopy, co-immunoprecipitation, GST pull-down, siRNA knockdown, Western blot for phosphorylation/cleavage Experimental cell research Medium 16764853
2012 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. 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 The Journal of cell biology High 22778279
1998 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. Molecular cloning of translocation breakpoints, sequence analysis, Northern blot expression analysis Human molecular genetics Medium 9618176
2009 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. Retroviral insertional mutagenesis screen, conditional Runx1 knockout mouse model, retroviral EVI5 overexpression rescue experiment Blood Medium 20008790
2020 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. Co-immunoprecipitation, CRISPR knockout, luciferase transcriptional reporter assay, flow cytometry, xenograft mouse model Cancer cell international Medium 32047362
2020 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. Co-immunoprecipitation, siRNA knockdown, wound healing/transwell invasion assays, xenograft mouse model Journal of experimental & clinical cancer research : CR Low 32393392
2015 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. Fine mapping/meta-analysis, co-immunoprecipitation followed by mass spectrometry, structural modeling of coiled-coil domain Human molecular genetics Medium 26433934
2024 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. RNAi depletion in Drosophila prothoracic gland, electron microscopy of vesicle morphology, ferritin co-immunoprecipitation/physical interaction assay, ferritin injection rescue experiment Nature communications Medium 38744835
2025 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. Co-immunoprecipitation, CRISPR knockout, overexpression, Western blot Scientific reports Low 41258404
2025 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. CRISPR knockout, overexpression, Western blot, ERK1/2 inhibitor functional rescue, immunofluorescence, qRT-PCR Current cancer drug targets Low 40525449
2022 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. Morpholino antisense knockdown in Xenopus tadpoles, RNA-sequencing, proliferation and apoptosis assays, Kdm6b/Kdm7a knockdown epistasis Frontiers in cell and developmental biology Medium 36605717

Source papers

Stage 0 corpus · 27 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2009 Stem cell exhaustion due to Runx1 deficiency is prevented by Evi5 activation in leukemogenesis. Blood 84 20008790
2006 The evi5 oncogene regulates cyclin accumulation by stabilizing the anaphase-promoting complex inhibitor emi1. Cell 83 16439210
2015 MicroRNA-135b, a HSF1 target, promotes tumor invasion and metastasis by regulating RECK and EVI5 in hepatocellular carcinoma. Oncotarget 67 25537516
2008 EVI5 is a risk gene for multiple sclerosis. Genes and immunity 62 18401352
2007 Identification of Rab11 as a small GTPase binding protein for the Evi5 oncogene. Proceedings of the National Academy of Sciences of the United States of America 61 17229837
2012 Evi5 promotes collective cell migration through its Rab-GAP activity. The Journal of cell biology 54 22778279
2006 The EVI5 TBC domain provides the GTPase-activating protein motif for RAB11. Oncogene 50 17099728
1998 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. Human molecular genetics 48 9618176
1995 Evi-5, a common site of retroviral integration in AKXD T-cell lymphomas, maps near Gfi-1 on mouse chromosome 5. Journal of virology 44 7474133
2013 The Evi5 family in cellular physiology and pathology. FEBS letters 36 23669355
2010 Tag-SNP analysis of the GFI1-EVI5-RPL5-FAM69 risk locus for multiple sclerosis. European journal of human genetics : EJHG 24 20087403
2005 EVI5 is a novel centrosomal protein that binds to alpha- and gamma-tubulin. Genomics 23 16033705
1997 Proviral integrations at the Evi5 locus disrupt a novel 90 kDa protein with homology to the Tre2 oncogene and cell-cycle regulatory proteins. Oncogene 22 9070650
2006 EVI5 protein associates with the INCENP-aurora B kinase-survivin chromosomal passenger complex and is involved in the completion of cytokinesis. Experimental cell research 19 16764853
2020 EVI5 is an oncogene that regulates the proliferation and metastasis of NSCLC cells. Journal of experimental & clinical cancer research : CR 16 32393392
2015 A non-synonymous single-nucleotide polymorphism associated with multiple sclerosis risk affects the EVI5 interactome. Human molecular genetics 14 26433934
2009 The NBPF1 promoter has been recruited from the unrelated EVI5 gene before simian radiation. Molecular biology and evolution 14 19282512
2013 GFI1B, EVI5, MYB--additional genes that cooperate with the human BCL6 gene to promote the development of lymphomas. Blood cells, molecules & diseases 9 23910958
2024 Drosophila Evi5 is a critical regulator of intracellular iron transport via transferrin and ferritin interactions. Nature communications 8 38744835
2020 The Evi5 oncogene promotes laryngeal cancer cells proliferation by stabilizing c-Myc protein. Cancer cell international 8 32047362
2017 Ecotropic Viral Integration Site 5 (EVI5) variants are associated with multiple sclerosis in Iranian population. Multiple sclerosis and related disorders 6 29141798
2017 Ecotropic Viral Integration Site 5 (EVI5) expression analysis in multiple sclerosis patients. Human antibodies 4 29036808
2015 In vitro and in vivo characterization of the Rab11-GAP activity of Drosophila Evi5. Methods in molecular biology (Clifton, N.J.) 4 25800843
2025 Novel EVI5::BRAF Gene Fusion in Infantile Fibrosarcoma: A Case Report and Review of Literature. International journal of molecular sciences 2 39940949
2022 Evi5 is required for Xenopus limb and tail regeneration. Frontiers in cell and developmental biology 2 36605717
2025 EVI5 Drives Lung Adenocarcinoma Progression through Modulation of the ERK1/2-c-Myc Signaling Axis. Current cancer drug targets 0 40525449
2025 EVI5 unveils a role of the oncogene in modulating the Rab11/PD-L1 pathway in lung adenocarcinoma. Scientific reports 0 41258404

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