Late steps in cytoplasmic maturation of assembly-competent axonemal outer arm dynein in Chlamydomonas require interaction of ODA5 and ODA10 in a complex
Late steps in cytoplasmic maturation of assembly-competent axonemal outer arm dynein in Chlamydomonas require interaction of ODA5 and ODA10 in a complex
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DOI：
10.1091/mbc.e15-05-0317
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发表时间：
2015-10-15
影响因子：
3.3
通讯作者：
Mitchell, David R.
中科院分区：
文献类型：
--
作者：
Dean, Anudariya B.;Mitchell, David R.
轴突动力蛋白是一种多亚基酶，必须在细胞质中预先组装，通过鞭毛内运输运输到纤毛中，并结合到双重微管上的特定位点，在那里它们的活性促进微管滑动运动。外动力蛋白臂（oda）需要装配因子来协助其预装配、运输和附着于货物（特定的双a管位点）。在衣藻中，三个组装因子ODA5、ODA8和ODA10表现出遗传相互作用，并被认为在一个复合体中相互作用，但我们最近发现鞭毛ODA8不与ODA5或ODA10共同作用。在这里，我们发现ODA5和ODA10在细胞质和鞭毛提取物中相互依赖并共存于一个复合体中。免疫荧光和免疫电镜显示，鞭毛中ODA10严格定位于1号双偶体的近端区域，而衣藻中完全缺乏ODA10。对ODAs与轴突双体的体外结合的研究揭示了ODA5/ODA10组装复合物在ODAs细胞质成熟成一种可以结合双体微管的形式中的作用。
Axonemal dyneins are multisubunit enzymes that must be preassembled in the cytoplasm, transported into cilia by intraflagellar transport, and bound to specific sites on doublet microtubules, where their activity facilitates microtubule sliding-based motility. Outer dynein arms (ODAs) require assembly factors to assist their preassembly, transport, and attachment to cargo (specific doublet A-tubule sites). In Chlamydomonas, three assembly factors-ODA5, ODA8, and ODA10-show genetic interactions and have been proposed to interact in a complex, but we recently showed that flagellar ODA8 does not copurify with ODA5 or ODA10. Here we show that ODA5 and ODA10 depend on each other for stability and coexist in a complex in both cytoplasmic and flagellar extracts. Immunofluorescence and immuno-electron microscopy reveal that ODA10 in flagella localizes strictly to a proximal region of doublet number 1, which completely lacks ODAs in Chlamydomonas. Studies of the in vitro binding of ODAs to axonemal doublets reveal a role for the ODA5/ODA10 assembly complex in cytoplasmic maturation of ODAs into a form that can bind to doublet microtubules.