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Originally published In Press as doi:10.1074/jbc.M500358200 on May 10, 2005
J. Biol. Chem., Vol. 280, Issue 26, 24634-24641, July 1, 2005
Interaction of the Salmonella-containing Vacuole with the Endocytic Recycling System*
Adam C. Smith ¶,
Judith T. Cirulis||** ,
James E. Casanova ,
Marci A. Scidmore¶¶, and
John H. Brumell ||||
From the
Infection, Immunity, Injury, and Repair Program and the ||Cardiovascular Research Program, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada, the Departments of Medical Genetics and Microbiology and **Biochemistry, University of Toronto, Toronto, Ontario M5S 1A1, Canada, the  Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, Virginia 22908, and the ¶¶Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853
Upon entry of the pathogen Salmonella enterica serovar Typhimurium into host cells, the majority of bacteria reside in a membrane-bound compartment called the Salmonella-containing vacuole (SCV). Previous studies have established that the SCV transiently interacts with early endosomes but only acquires a subset of late endosomal/lysosomal proteins. However, the complete set of interactions between the SCV and the endocytic machinery has yet to be characterized. In this study, we have shown that four characterized regulators of endocytic recycling were present on the SCV after invasion. Interaction kinetics were different for each of the regulators; ARF6 and Rab4 associated immediately, but their presence was diminished 60 min post-infection, whereas syntaxin13 and Rab11 association peaked at 60 min. Using a dominant negative approach, we determined that Rab11 regulates the recycling of CD44 from the vacuole but had no effect on major histocompatibility complex (MHC) class I recycling. In contrast, syntaxin13 regulated the recycling of MHC class I but not of CD44. We also determined that maturation of the SCV, measured by the acquisition of lysosomal associated membrane protein-1, slowed when recycling was impaired. These findings suggest that protein movement through the endocytic recycling system is regulated through at least two concurrent pathways and that efficient interaction with these pathways is necessary for maturation of the Salmonella-containing vacuole. We also demonstrate the utility of using Salmonella invasion as a model of endosomal recycling events.
Received for publication, January 11, 2005
, and in revised form, May 4, 2005.
* Infrastructure for the Brumell Laboratory was provided by a New Opportunities Fund from the Canadian Foundation for Innovation and the Ontario Innovation Trust. 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.
The on-line version of this article (available at http://www.jbc.org) contains supplemental material in the form of a movie depicting the accumulation of EGFP-Rab11 around the SCV upon invasion.
¶ Recipient of a University of Toronto open scholarship, a studentship from the Natural Sciences and Engineering Research Council of Canada, and the Toronto Star student bursary through The Hospital for Sick Children Research Training Centre.
 Supported by the Samuel Lunenfeld Summer Student Program.
|||| Recipient of a New Investigator award from the Canadian Institutes of Health Research and the Premier's Research Excellence Award from the Ontario Ministry of Economic Development and Trade and to whom correspondence should be addressed: IIIR Program, Hospital for Sick Children, 555 University Ave., Toronto, Ontario M5G 1X8, Canada. Tel.: 416-813-7654 (ext. 3555); Fax: 416-813-5028; E-mail: john.brumell{at}sickkids.ca.

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Copyright © 2005 by the American Society for Biochemistry and Molecular Biology.
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