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Vol. 16, Issue 4, 1640-1650, April 2005
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* Unité de Morphogenèse et Signalisation Cellulaires, Institut Curie, Centre National de la Recherche Scientifique Unité Mixte de Recherche 144, 75248 Paris Cedex 05, France;
Cell and Molecular Biology, Division of Biomedical Sciences, Faculty of Medicine, Imperial College London, London SW7 2AZ, United Kingdom; and
Department of Basic Medical Sciences, St. George's Hospital Medical School, London SW17 ORE, United Kingdom
Submitted September 3, 2004;
Revised January 11, 2005;
Accepted January 12, 2005
Monitoring Editor: Suzanne Pfeffer
Rab GTPases have been implicated in the regulation of specific microtubule- and actin-based motor proteins. We devised an in vitro motility assay reconstituting the movement of melanosomes on actin bundles in the presence of ATP to investigate the role of Rab proteins in the actin-dependent movement of melanosomes. Using this assay, we confirmed that Rab27 is required for the actin-dependent movement of melanosomes, and we showed that a second Rab protein, Rab8, also regulates this movement. Rab8 was partially associated with mature melanosomes. Expression of Rab8Q67L perturbed the cellular distribution and increased the frequency of microtubule-independent movement of melanosomes in vivo. Furthermore, anti-Rab8 antibodies decreased the number of melanosomes moving in vitro on actin bundles, whereas melanosomes isolated from cells expressing Rab8Q67L exhibited 70% more movements than wild-type melanosomes. Together, our observations suggest that Rab8 is involved in regulating the actin-dependent movement of melanosomes.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Address correspondence to: Evelyne Coudrier (coudrier{at}curie.fr).
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