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Infection and Immunity, June 1999, p. 3002-3008, Vol. 67, No. 6
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Effects of Cytotoxic Necrotizing Factor 1 and Lethal Toxin on Actin Cytoskeleton and VE-Cadherin Localization in Human Endothelial Cell Monolayers

Valérie Vouret-Craviari,1 Dominique Grall,1 Gilles Flatau,2 Jacques Pouysségur,1 Patrice Boquet,2 and Ellen Van Obberghen-Schilling1,*

Centre de Biochimie, CNRS UMR 6543, 06108 Nice Cedex 2,1 and INSERM U452, Faculté de Médecine de Nice, 06107 Nice Cedex 2,2 France

Received 19 January 1999/Returned for modification 4 March 1999/Accepted 24 March 1999

Integrity of the vascular endothelium is largely dependent on endothelial cell shape and establishment of intercellular junctions. Certain pathogenic bacterial toxins alter the cytoskeletal architecture of intoxicated cells by modulating the GTPase activity of p21 Rho family proteins. In the present study we have analyzed the effect of Rho-directed toxins on the actin cytoskeleton and monolayer integrity of endothelial cells. We report here that Escherichia coli cytotoxic necrotizing factor 1 (CNF1) activates Rho in human umbilical vein endothelial cells (HUVEC). In confluent monolayers, CNF1 treatment induces prominent stress fiber formation without significantly modifying peripheral localization of VE-cadherin, a specific marker of vascular endothelial cell adherens junctions. Further, Rho activation with CNF1 blocks thrombin-induced redistribution of VE-cadherin staining and gap formation in HUVEC monolayers. Inhibition of Rho by prolonged treatment of cells with C3 exoenzyme (Clostridium botulinum) eliminates actin stress fibers without disrupting the continuity of VE-cadherin staining, indicating that Rho-dependent stress fibers are not required for maintaining this adhesion receptor at sites of intercellular contact. Lethal toxin (Clostridium sordellii), an inhibitor of Rac as well as Ras and Rap, potently disrupts the actin microfilament system and monolayer integrity in HUVEC cultures.

* Corresponding author. Mailing address: Centre de Biochimie, CNRS UMR6543, Parc Valrose, 06108 Nice Cedex 2, France. Phone: 33.492 07 6425. Fax: 33.492 07 6432. E-mail: vanobber{at}unice.fr.

Infection and Immunity, June 1999, p. 3002-3008, Vol. 67, No. 6
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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