Alternative Splicing in Class V Myosins Determines Association with Rab10*
- Departments of Surgery and Cell and Developmental Biology, Vanderbilt University School of Medicine, and the Nashville Veterans Affairs Medical Center, Nashville, Tennessee 37232-2733
- 1 To whom correspondence should be addressed: Vanderbilt University School of Medicine, Dept. of Surgery, Epithelial Biology Program, 4160A MRB III, 465 21st St. S., Nashville, TN 37232-2733. Tel.: 615-936-3726; Fax: 615-343-1591; E-mail: jim.goldenring{at}vanderbilt.edu.
Abstract
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.
Footnotes
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↵2 The abbreviations used are: EGFP, enhanced green fluorescent protein; FRET, fluorescence resonance energy transfer.
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↵* This work was supported, in whole or in part, by National Institutes of Health Grants R01 DK070856 (NIDDK) and R01 DK48370 (to J. R. G.) and F32 DK072789 (to J. T. R.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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The on-line version of this article (available at http://www.jbc.org) contains supplemental Fig. 1 and 2 and Movies 1-3.
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- Received August 1, 2008.
- Revision received October 22, 2008.
- The American Society for Biochemistry and Molecular Biology, Inc.
