Analysis of the dynein-dynactin interaction in vitro and in vivo
Analysis of the dynein-dynactin interaction in vitro and in vivo
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DOI：
10.1091/mbc.e03-01-0025
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发表时间：
2003-12-01
影响因子：
3.3
通讯作者：
Schroer, TA
中科院分区：
文献类型：
--
作者：
King, SJ;Brown, CL;Schroer, TA
细胞质动力蛋白和动力肌动蛋白是巨道尔顿大小的多亚基分子，它们共同起细胞骨架马达的作用。在本研究中，我们探讨了动力蛋白-动力肌动蛋白在体外结合的机制，然后将我们的研究结果扩展到体内环境。溶液结合试验用于确定动力蛋白中间链（IC）和动力蛋白p150（Glued）亚基中的结合结构域。动力蛋白IC的一系列片段的瞬时过表达用于确定该亚基在哺乳动物组织培养细胞中对动力蛋白功能的重要性。我们的研究结果表明，一个功能性的动力蛋白-dynactin相互作用是必要的适当的微管组织和运输和定位的中心体组件和内膜室。动力蛋白IC片段对内膜定位有不同的影响，表明不同的内膜可能通过不同的机制结合动力蛋白。
Cytoplasmic dynein and dynactin are megadalton-sized multisubunit molecules that function together as a cytoskeletal motor. In the present study, we explore the mechanism of dynein-dynactin binding in vitro and then extend our findings to an in vivo context. Solution binding assays were used to define binding domains in the dynein intermediate chain (IC) and dynactin p150(Glued) subunit. Transient overexpression of a series of fragments of the dynein IC was used to determine the importance of this subunit for dynein function in mammalian tissue culture cells. Our results suggest that a functional dynein-dynactin interaction is required for proper microtubule organization and for the transport and localization of centrosomal components and endomembrane compartments. The dynein IC fragments have different effects on endomembrane localization, suggesting that different endomembranes may bind dynein via distinct mechanisms.