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通过绿色荧光蛋白筛选策略鉴定一种调节微管组织和胞质分裂的新型微管相关蛋白。

Identification of a novel microtubule-associated protein that regulates microtubule organization and cytokinesis by using a GFP-screening strategy.

作者信息

Manabe Ri ichiroh, Whitmore Leanna, Weiss Jonathan M, Horwitz Alan Rick

机构信息

Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.

出版信息

Curr Biol. 2002 Nov 19;12(22):1946-51. doi: 10.1016/s0960-9822(02)01299-x.

Abstract

Microtubules play critical roles in a variety of cell processes, including mitosis, organelle transport, adhesion and migration, and the maintenance of cell polarity. Microtubule-associated proteins (MAPs) regulate the dynamic organization and stability of microtubules, often through either cell-specific or cell division stage-specific interactions. To identify novel cytoskeletal-associated proteins and peptides that regulate microtubules and other cytoskeletal and adhesive structures, we have developed a GFP cDNA screening strategy based on identifying gene products that localize to these structures. Using this approach, we have identified a novel MAP, GLFND, that shows homology to the Opitz syndrome gene product [6], localizes to a subpopulation of microtubules that are acetylated, and protects microtubules from depolymerization with nocodazole. Expression of an N-terminal deletion binds microtubules but alters their organization. During the cell cycle, GLFND dissociates from microtubules at the beginning of mitosis and then reassociates at cytokinesis. Furthermore, ectopic expression of GLFND inhibits cell division and cytokinesis in CHO cells. These observations make GLFND unique among MAPs characterized thus far.

摘要

微管在多种细胞过程中发挥关键作用,包括有丝分裂、细胞器运输、黏附与迁移以及细胞极性的维持。微管相关蛋白(MAPs)通常通过细胞特异性或细胞分裂阶段特异性相互作用来调节微管的动态组织和稳定性。为了鉴定调节微管以及其他细胞骨架和黏附结构的新型细胞骨架相关蛋白和肽段,我们基于鉴定定位于这些结构的基因产物,开发了一种绿色荧光蛋白(GFP)cDNA筛选策略。利用这种方法,我们鉴定出一种新型微管相关蛋白GLFND,它与奥匹茨综合征基因产物具有同源性[6],定位于乙酰化的微管子群体,并保护微管不被诺考达唑解聚。N端缺失的GLFND表达产物能结合微管,但会改变其组织结构。在细胞周期中,GLFND在有丝分裂开始时从微管上解离,然后在胞质分裂时重新结合。此外,GLFND的异位表达会抑制CHO细胞的细胞分裂和胞质分裂。这些观察结果使得GLFND在迄今为止所鉴定的微管相关蛋白中独具特色。

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