EVI5 is a novel centrosomal protein that binds to α- and γ-tubulin

Faitar, Silviu L.; Dabbeekeh, Jeremy; Ranalli, Tamara A.; Cowell, John K.

doi:10.1016/j.ygeno.2005.06.002

Cited by 23 publications

(25 citation statements)

References 43 publications

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“…The identification of GTPase activator proteins (GAPs) and GTPase exchange factors for each Rab protein, therefore, is critical for understanding the mechanisms that regulate their enzymatic activity and their biochemical function in the cell. To identify proteins that interact with the EVI5 TBC domain, a construct containing the N-terminal 445 amino acids of EVI5 (Faitar et al, 2005) was cloned into the pGEX-5X-1 vector and expressed in Escherichia coli (BL21) as a gluthathione-S-transferase (GST) fusion protein. The bacterially expressed protein was immobilized on glutathione-agarose beads and used in pulldown experiments with total protein extracts from IMR5 neuroblastoma cells.…”

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“…The human EVI5 protein, which was identified at the breakpoint in a constitutional chromosome translocation in a patient with stage 4S neuroblastoma (Roberts et al, 1998), contains a TBC domain near its N terminus. EVI5 has been identified in the centrosome in interphase cells (Faitar et al, 2005) and in the midbody during the terminal stages of cytokinesis. Small interfering RNA knockdown of EVI5 results in multinucleate cells because of an inability of daughter cell abscission (Faitar et al, 2006).…”

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confidence: 99%

“…In addition to the GST bait and the EVI5 protein itself, this analysis identified Rab11 (represented by two different peptides coving B9% of the protein) as a potentially interacting protein (Figure 1). To confirm this interaction, the endogenous EVI5 protein was immunoprecipitated (IP) from IMR5 and HeLa cell lysates, using the EVI5 antibody described previously (Faitar et al, 2005). The presence of Rab11 among the proteins IP with this antibody was analysed using Western blotting with an anti-Rab11 antibody.…”

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The EVI5 TBC domain provides the GTPase-activating protein motif for RAB11

et al. 2006

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The human EVI5 gene was originally isolated through its involvement with a constitutional chromosome translocation in a patient with stage 4S neuroblastoma. Recently, it has been shown that EVI5 is a centrosomal protein in interphase cells, which relocalizes to the midbody during late phases of mitosis. Disruption of its function leads to incomplete cell division and the formation of multinucleate cells. The EVI5 protein contains a TBC (TRE2/BUB/ CDC16 homology) motif located in the N-terminal region. Proteins containing a TBC domain have been shown in some cases to act as GTPase-activating proteins (GAPs) and function through the interaction with Rab-like small G proteins. Despite the identification of over 50 TBCcontaining proteins, and over 70 Rab-like proteins, only three combinations have been shown to have Rab/GAP activity to date. In this study, using linear ion trap mass spectroscopy, we have demonstrated that EVI5 exists in a protein complex with Rab11. Further, using a specific Rab-binding assay, we have shown that EVI5 preferentially interacts with the guanosine triphosphate-bound form of Rab11, and in a GAP activity assay, we have confirmed that EVI5 functions as a GAP for the Rab11 GTPase.Oncogene ( Proteins with homology to the so-called TBC domain, consisting of an B200-amino-acid motif initially identified in the TRE2/BUB2/CDC16 genes (Richardson and Zon, 1995), are considered to function as GTPaseactivating proteins (GAPs), partnering with Rab-like small G proteins. Rab GTPases are members of the Ras superfamily of guanosine triphosphate (GTP)-binding proteins that play critical roles in the regulation of important membrane and protein trafficking events in the cell. Many of the Rab proteins are associated with fundamental biological processes such as vesicle fusion, receptor recycling, membrane transport and cytokinesis (Zerial and McBride, 2001). There are, however, over 50 human proteins with predicted TBC domains, and over 70 human Rab proteins, of which only two TBC domain-containing proteins have so far been shown to demonstrably have Rab/GAP activity. The human EVI5 protein, which was identified at the breakpoint in a constitutional chromosome translocation in a patient with stage 4S neuroblastoma (Roberts et al., 1998), contains a TBC domain near its N terminus. EVI5 has been identified in the centrosome in interphase cells (Faitar et al., 2005) and in the midbody during the terminal stages of cytokinesis. Small interfering RNA knockdown of EVI5 results in multinucleate cells because of an inability of daughter cell abscission (Faitar et al., 2006). Because of the essential role of EVI5 in cytokinesis, as well as its implicated involvement in cancer development, we used a proteomics approach and identified the Rab protein that is activated by the EVI5 TBC domain.Rab proteins function by cycling between the biologically active GTP-bound form and the guanosine diphosphate (GDP)-bound inactive form. In the active GTP-bound conformation, these proteins can directly interact with specific eff...

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The EVI5 TBC domain provides the GTPase-activating protein motif for RAB11

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“…21 EVI-5 is also a nuclear zinc-finger protein that functions as a transcriptional repressor that is possibly involved in centrosome stability and dynamics. 22 Interestingly, SNP4 located in exon 15 of the EVI5 gene is a nonsynonymous polymorphism (612 His/Gln), but the potential effect on the EVI5N domain is unknown. Statistically, SNP4 did not survive Benjamini-Hochberg correction.…”

Section: Discussionmentioning

confidence: 99%

Tag-SNP analysis of the GFI1-EVI5-RPL5-FAM69 risk locus for multiple sclerosis

Alcina

Fernández²,

González

et al. 2010

Eur J Hum Genet

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A recent genome-wide association study conducted by the International Multiple Sclerosis Genetic Consortium (IMSGC) identified, among others, a number of putative multiple sclerosis (MS) susceptibility variants at position 1p22. Twenty-one SNPs positively associated with MS were located at the GFI-EVI5-RPL5-FAM69A locus. In this study, we performed an analysis and fine mapping of this locus, genotyping eight Tag-SNPs in 732 MS patients and 974 controls from Spain. We observed an association with MS in three of eight Tag-SNPs: rs11804321 (P¼0.008, OR¼1.29; 95% CI¼1.08-1.54), rs11808092 (P¼0.048, OR¼1.19; 95% CI¼1.03-1.39) and rs6680578 (P¼0.0082, OR¼1.23; 95% CI¼1.07-1.41). After correcting for multiple comparisons and using logistic regression analysis to test the addition of each SNP to the most associated SNPs, we observed that rs11804321 alone was sufficient to model the association. This Tag-SNP captures two SNPs in complete linkage disequilibrium (r 2 ¼1), both located within the 17th intron of the EVI5 gene. Our findings agree with the corresponding data of the recent IMSGC study and present new genetic evidence that points to EVI5 as a factor of susceptibility to MS.

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“…EVI5 is a centrosomal protein during interphase, but it relocalises to the midbody during late phases of mitosis. Its disruption leads to incomplete cell division and formation of multinucleate cells (Faitar et al, 2005). EVI5 also exists in a protein complex with GTPbound Rab11 and functions as a GTPase-activating protein for this small GTPase (Dabbeekeh et al, 2007).…”

Section: Nbpf Promoter Analysismentioning

confidence: 99%