Alternative Splicing in Class V Myosins Determines Association with Rab10
Alternative Splicing in Class V Myosins Determines Association with Rab10
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DOI：
10.1074/jbc.m805957200
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发表时间：
2009-01-09
影响因子：
4.8
通讯作者：
Goldenring, James R.
中科院分区：
文献类型：
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作者：
Roland, Joseph T.;Lapierre, Lynne A.;Goldenring, James R.
Rab蛋白通过调节蛋白复合物的组装影响囊泡运输途径。先前的研究已经证明Rab11a和Rab8a可以与肌球蛋白Vb尾部区域相互作用并调节不同的转运途径。我们现在已经确定一种相关的Rab蛋白Rab10可以与肌凝蛋白Va、肌凝蛋白Vb和肌凝蛋白Vc相互作用。Rab10定位于与Rab8a部分重叠的小管和囊泡系统。Rab8a和Rab10均因myosin V尾部的显性阴性表达而错定位。与Rab10的相互作用依赖于在肌凝蛋白Va和肌凝蛋白Vb中存在选择性剪接的外显子D，以及在肌凝蛋白Vc中存在同源区域。酵母双杂交实验和荧光共振能量转移研究证实，Rab10在体内与肌凝蛋白V尾部结合需要选择性剪接的外显子d。与我们之前的工作相反，我们发现Rab11a可以与肌凝蛋白Va和肌凝蛋白Vb尾部相互作用，而不依赖于它们的剪接异构体。这些结果表明，Rab gtpase通过募集多种肌球蛋白V亚型来调节多种内吞运输途径。
Rab proteins influence vesicle trafficking pathways through the assembly of regulatory protein complexes. Previous investigations have documented that Rab11a and Rab8a can interact with the tail region of myosin Vb and regulate distinct trafficking pathways. We have now determined that a related Rab protein, Rab10, can interact with myosin Va, myosin Vb, and myosin Vc. Rab10 localized to a system of tubules and vesicles that have partially overlapping localization with Rab8a. Both Rab8a and Rab10 were mislocalized by the expression of dominant-negative myosin V tails. Interaction with Rab10 was dependent on the presence of the alternatively spliced exon D in myosin Va and myosin Vb and the homologous region in myosin Vc. Yeast two-hybrid assays and fluorescence resonance energy transfer studies confirmed that Rab10 binding to myosin V tails in vivo required the alternatively spliced exon D. In contrast to our previous work, we found that Rab11a can interact with both myosin Va and myosin Vb tails independent of their splice isoform. These results indicate that Rab GTPases regulate diverse endocytic trafficking pathways through recruitment of multiple myosin V isoforms.