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Infection and Immunity, May 2006, p. 2552-2561, Vol. 74, No. 5
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.5.2552-2561.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A C-Terminal Domain Targets the Pseudomonas aeruginosa Cytotoxin ExoU to the Plasma Membrane of Host Cells

Shira D. P. Rabin,1,{dagger} Jeffrey L. Veesenmeyer,1 Kathryn T. Bieging,1 and Alan R. Hauser1,2*

Departments of Microbiology/Immunology,1 Medicine, Northwestern University, Chicago, Illinois 606112

Received 29 July 2005/ Returned for modification 20 September 2005/ Accepted 8 February 2006

ExoU, a phospholipase injected into host cells by the type III secretion system of Pseudomonas aeruginosa, leads to rapid cytolytic cell death. Although the importance of ExoU in infection is well established, the mechanism by which this toxin kills host cells is less clear. To gain insight into how ExoU causes cell death, we examined its subcellular localization following transfection or type III secretion/translocation into HeLa cells. Although rapid cell lysis precluded visualization of wild-type ExoU by fluorescence microscopy, catalytically inactive toxin was readily detected at the periphery of HeLa cells. Biochemical analysis confirmed that ExoU was targeted to the membrane fraction of transfected cells. Visualization of ExoU peptides fused with green fluorescent protein indicated that the domain responsible for this targeting was in the C terminus of ExoU, between residues 550 and 687. Localization to the plasma membrane occurred within 1 h of expression, which is consistent with the kinetics of cytotoxicity. Together, these results indicate that a domain between residues 550 and 687 of ExoU targets this toxin to the plasma membrane, a process that may be important in cytotoxicity.

* Corresponding author. Mailing address: Department of Microbiology/Immunology, Northwestern University, 303 East Chicago Ave., Searle 6-495, Chicago, IL 60611. Phone: (312) 503-1044. Fax: (312) 503-1339. E-mail: ahauser{at}northwestern.edu.

Editor: J. B. Bliska

{dagger} Present address: Center for Oral Health & Systemic Disease, School of Dentistry, University of Louisville, Louisville, KY 40292.

Infection and Immunity, May 2006, p. 2552-2561, Vol. 74, No. 5
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.5.2552-2561.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sutterwala, F. S., Mijares, L. A., Li, L., Ogura, Y., Kazmierczak, B. I., Flavell, R. A. (2007). Immune recognition of Pseudomonas aeruginosa mediated by the IPAF/NLRC4 inflammasome. JEM 204: 3235-3245 [Abstract] [Full Text]  
  • Zhang, Y., Deng, Q., Barbieri, J. T. (2007). Intracellular Localization of Type III-delivered Pseudomonas ExoS with Endosome Vesicles. J. Biol. Chem. 282: 13022-13032 [Abstract] [Full Text]  


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