Faitar Silviu L, Dabbeekeh Jeremy T S, Ranalli Tamara A, Cowell John K
Department of Cancer Genetics, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Genomics. 2005 Nov;86(5):594-605. doi: 10.1016/j.ygeno.2005.06.002. Epub 2005 Jul 19.
The human EVI5 protein carries a TBC domain indicative of Rab GTPase activating protein (GAP) activity, and an extensive coiled-coil motif in the C-terminal region. EVI5 is ubiquitously expressed in adult, fetal, and cancer tissues and exists as two mRNA species resulting from differential use of polyadenylation signals. Western blot analysis suggests that different molecular weight protein species are probably generated by posttranslational modification. FPLC analysis demonstrates that EVI5 protein can form dimers and confocal microscopy indicates that EVI5, in addition to a diffuse localization in the nucleus, also preferentially localizes to the pericentriolar material in interphase cells. Immunoprecipitation and GST pull-down experiments demonstrate that EVI5 exists in complexes with both alpha- and gamma-tubulin. Both interactions are localized to the N-terminal part of the EVI5 protein. Thus, EVI5 is a novel centrosomal protein with a complex expression pattern and subcellular localization, possibly involved in centrosome stability and dynamics.
人类EVI5蛋白带有一个表明Rab GTP酶激活蛋白（GAP）活性的TBC结构域，以及在C端区域的一个广泛的卷曲螺旋基序。EVI5在成人、胎儿和癌组织中普遍表达，并以两种mRNA形式存在，这是由多聚腺苷酸化信号的差异使用导致的。蛋白质免疫印迹分析表明，不同分子量的蛋白质种类可能是由翻译后修饰产生的。快速蛋白质液相色谱分析表明，EVI5蛋白可以形成二聚体，共聚焦显微镜显示，EVI5除了在细胞核中呈弥散定位外，还优先定位于间期细胞的中心粒周围物质。免疫沉淀和谷胱甘肽-S-转移酶下拉实验表明，EVI5与α-微管蛋白和γ-微管蛋白均以复合物形式存在。这两种相互作用都定位于EVI5蛋白的N端部分。因此，EVI5是一种新型的中心体蛋白，具有复杂的表达模式和亚细胞定位，可能参与中心体的稳定性和动态变化。
