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Molecular and Cellular Biology, October 2006, p. 7299-7317, Vol. 26, No. 19
0270-7306/06/$08.00+0     doi:10.1128/MCB.02405-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Bioinformatic and Comparative Localization of Rab Proteins Reveals Functional Insights into the Uncharacterized GTPases Ypt10p and Ypt11p{dagger}

Stéphanie Buvelot Frei,1 Peter B. Rahl,2 Maria Nussbaum,3 Benjamin J. Briggs,4 Monica Calero,2 Stephanie Janeczko,5 Andrew D. Regan,6 Catherine Z. Chen,2 Yves Barral,1 Gary R. Whittaker,8 and Ruth N. Collins7*

Institut für Biochemie, Swiss Federal Institute of Technology (ETH), CH-8093 Zürich, Switzerland,1 Graduate Program in Pharmacology, Cornell University, Ithaca, New York 14853,2 Research Apprenticeship in Biological Sciences, Cornell University Summer College, Ithaca, New York 14853,3 Honors Program in Undergraduate Biology, Cornell University, Ithaca, New York 14853,4 Leadership Program for Veterinary Students, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853,5 Graduate Program in Microbiology, Cornell University, Ithaca, New York 14853,6 Departments of Molecular Medicine,7 Microbiology and Immunology, Cornell University, Ithaca, New York 148538

Received 18 December 2005/ Returned for modification 21 February 2006/ Accepted 11 July 2006

A striking characteristic of a Rab protein is its steady-state localization to the cytosolic surface of a particular subcellular membrane. In this study, we have undertaken a combined bioinformatic and experimental approach to examine the evolutionary conservation of Rab protein localization. A comprehensive primary sequence classification shows that 10 out of the 11 Rab proteins identified in the yeast (Saccharomyces cerevisiae) genome can be grouped within a major subclass, each comprising multiple Rab orthologs from diverse species. We compared the locations of individual yeast Rab proteins with their localizations following ectopic expression in mammalian cells. Our results suggest that green fluorescent protein-tagged Rab proteins maintain localizations across large evolutionary distances and that the major known player in the Rab localization pathway, mammalian Rab-GDI, is able to function in yeast. These findings enable us to provide insight into novel gene functions and classify the uncharacterized Rab proteins Ypt10p (YBR264C) as being involved in endocytic function and Ypt11p (YNL304W) as being localized to the endoplasmic reticulum, where we demonstrate it is required for organelle inheritance.


* Corresponding author. Mailing address: Department of Molecular Medicine, Cornell University, Ithaca, NY 14853-6401. Phone: (607) 253-4123. Fax: (607) 253-3659. E-mail: rnc8{at}cornell.edu.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.


Molecular and Cellular Biology, October 2006, p. 7299-7317, Vol. 26, No. 19
0270-7306/06/$08.00+0     doi:10.1128/MCB.02405-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




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