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MBC in Press, published online ahead of print May 16, 2007
Mol. Biol. Cell 10.1091/mbc.E07-02-0169

A more recent version of this article appeared on August 1, 2007
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Submitted on February 23, 2007
Revised on April 16, 2007
Accepted on May 7, 2007

Myosin Vb Interacts with Rab8a on a Tubular Network Containing EHD1 and EHD3

Joseph T. Roland,* Anne K. Kenworthy,{dagger}{ddagger} Johan Peranen,{sect} Steve Caplan,|| and James R. Goldenring*{dagger}

Departments of *Surgery, {dagger}Cell and Developmental Biology, and {ddagger}Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232-2733; Nashville Veterans Affairs Medical Center, Nashville, TN 37212-2637; {sect}Institute of Biotechnology, University of Helsinki, FIN-00014 Helsinki, Finland; ||Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE 68198

Monitoring Editor: Patrick Brennwald

Cells utilize multiple pathways to internalize and recycle cell surface components. While Rab11a and Myosin Vb are involved in the recycling of proteins internalized by clathrin-mediated endocytosis, Rab8a has been implicated in nonclathrin-dependent endocytosis and recycling. By yeast two-hybrid assays, we have now demonstrated that Myosin Vb can interact with Rab8a, but not Rab8b. We have confirmed the interaction of Myosin Vb with Rab11a and Rab8a in vivo using fluorescent resonant energy transfer (FRET) techniques. Rab8a and Myosin Vb colocalize to a tubular network containing EHD1 and EHD3, which does not contain Rab11a. Myosin Vb tail can cause the accumulation of both Rab11a and Rab8a in collapsed membrane cisternae, while dominant-negative Rab11-FIP2(129-512) selectively accumulates Rab11a, but not Rab8a. Additionally, dynamic live cell imaging demonstrates distinct pathways for Rab11a and Rab8a vesicle trafficking. These findings indicate that Rab8a and Rab11a define different recycling pathways that both employ Myosin Vb.

Address correspondence to: James R. Goldenring (jim.goldenring{at}vanderbilt.edu)




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