This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Isaksson, E. L.
Right arrow Articles by Wolf-Watz, H.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Isaksson, E. L.
Right arrow Articles by Wolf-Watz, H.

 Previous Article  |  Next Article 

Infection and Immunity, November 2009, p. 4740-4749, Vol. 77, No. 11
0019-9567/09/$08.00+0     doi:10.1128/IAI.00333-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The Membrane Localization Domain Is Required for Intracellular Localization and Autoregulation of YopE in Yersinia pseudotuberculosis{triangledown}

Elin L. Isaksson, Margareta Aili,{dagger} Anna Fahlgren, Sara E. Carlsson, Roland Rosqvist, and Hans Wolf-Watz*

Department of Molecular Biology, Laboratory of Molecular Infectious Medicine, Umeå University, S-90187 Umeå, Sweden

Received 23 March 2009/ Returned for modification 27 April 2009/ Accepted 5 August 2009

Recent work has shown that a domain of YopE of Yersinia pseudotuberculosis ranging from amino acids 54 to 75 (R. Krall, Y. Zhang, and J. T. Barbieri, J. Biol. Chem. 279:2747-2753, 2004) is required for proper localization of YopE after ectopic expression in eukaryotic cells. This domain, called the membrane localization domain (MLD), has not been extensively studied in Yersinia. Therefore, an in cis MLD deletion mutant of YopE was created in Y. pseudotuberculosis. The mutant was found to secrete and translocate YopE at wild-type levels. However, the mutant was defective in the autoregulation of YopE expression after the infection of HeLa cells. Although the mutant translocated YopE at wild-type levels, it showed a delayed HeLa cell cytotoxicity. This delay was not caused by a change in GTPase activating protein (GAP) activity, since the mutant showed wild-type YopE GAP activity toward Rac1 and RhoA. The MLD mutant displayed a changed intracellular localization pattern of YopE in HeLa cells after infection, and the YopE{Delta}MLD protein was found to be dispersed within the whole cell, including the nucleus. In contrast, wild-type YopE was found to localize to the perinuclear region of the cell and was not found in the nucleus. In addition, the yopE{Delta}MLD mutant was avirulent. Our results suggest that YopE must target proteins other than RhoA and Rac1 and that the MLD is required for the proper targeting and hence virulence of YopE during infection. Our results raise the question whether YopE is a regulatory protein or a "true" virulence effector protein.

* Corresponding author. Mailing address: Department of Molecular Biology, Laboratory of Molecular Infectious Medicine, Umeå University, S-90187 Umeå, Sweden. Phone: 46907856753. Fax: 4690772630. E-mail: hans.wolf-watz{at}molbiol.umu.se

{triangledown} Published ahead of print on 17 August 2009.

Editor: J. B. Bliska

{dagger} Present address: Department of Microbiology and Immunology, Melbourne University, Parkville, Victoria 3010, Australia.

Infection and Immunity, November 2009, p. 4740-4749, Vol. 77, No. 11
0019-9567/09/$08.00+0     doi:10.1128/IAI.00333-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»