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Originally published In Press as doi:10.1074/jbc.C000153200 on March 21, 2000
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J Biol Chem, Vol. 275, Issue 19, 14009-14012, May 12, 2000

ACCELERATED PUBLICATION
Cell Vacuolation Induced by the VacA Cytotoxin of Helicobacter pylori Is Regulated by the Rac1 GTPase*

Neil A. HotchinDagger §, Timothy L. Cover, and Nasreen AkhtarDagger

From the Dagger  School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom and the  Division of Infectious Disease, Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605

Chronic gastric infection with the Gram-negative bacterium Helicobacter pylori is a major contributing factor in the development of duodenal ulcers and is believed to be a significant risk factor in the development of gastric tumors. The VacA cytotoxin of H. pylori is a 90-kDa secreted protein that forms trans-membrane ion channels. In epithelial cells, VacA activity is associated with the rapid formation of acidic vacuoles enriched for late endosomal and lysosomal markers. Rac1 is a member of the Rho family of small GTP-binding proteins that regulate reorganization of the actin cytoskeleton and intracellular signal transduction and are being shown increasingly to play a role in membrane trafficking events. In this study we report that: (i) green fluorescent-tagged Rac1 localizes around the perimeter of the vacuoles induced by VacA; (ii) expression of dominant negative Rac1 in epithelial cells inhibits vacuole formation; (iii) expression of constitutively active Rac1 potentiates the activity of VacA. Taken together, these data demonstrate a role for Rac1 in the regulation of VacA activity.

* This work was supported by Medical Research Council Project Grant G9537491 (to N. A. H.) and by grants from the National Institutes of Health (R01 AI-39657 and DK-53623) and the Department of Veterans Affairs (to T. L. C.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ To whom correspondence should be addressed. E-mail: n.a.hotchin@ bham.ac.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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