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
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Garrity-Ryan, L.
Right arrow Articles by Engel, J. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Garrity-Ryan, L.
Right arrow Articles by Engel, J. N.
Right arrowPubmed/NCBI databases
*Compound via MeSH
*Substance via MeSH
Hazardous Substances DB
*(L)-ARGININE

 Previous Article  |  Next Article 

Infection and Immunity, December 2000, p. 7100-7113, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

The Arginine Finger Domain of ExoT Contributes to Actin Cytoskeleton Disruption and Inhibition of Internalization of Pseudomonas aeruginosa by Epithelial Cells and Macrophages

L. Garrity-Ryan, B. Kazmierczak, R. Kowal, J. Comolli,dagger A. Hauser,Dagger and J. N. Engel*

Department of Medicine, Department of Microbiology and Immunology, and the Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California 94143

Received 1 June 2000/Returned for modification 12 July 2000/Accepted 22 September 2000

Pseudomonas aeruginosa, an important nosocomial pathogen of humans, expresses a type III secretion system that is required for virulence. Previous studies demonstrated that the lung-virulent strain PA103 has the capacity to be either cytotoxic or invasive. Analyses of mutants suggest that PA103 delivers a negative regulator of invasion, or anti-internalization factor, to host cells via a type III secretion system. In this work we show that the type III secreted protein ExoT inhibits the internalization of PA103 by polarized epithelial cells (Madin-Darby canine kidney cells) and J774.1 macrophage-like cells. ExoS, which is closely related to ExoT but has additional ADP-ribosylating activity, can substitute for ExoT as an anti-internalization factor. ExoT contains a signature arginine finger domain found in GTPase-activating proteins. Mutation of the conserved arginine in ExoT diminished its anti-internalization activity and altered its ability to disrupt the actin cytoskeleton. Cell fractionation experiments showed that ExoT is translocated into host cells and that mutation of the arginine finger did not disrupt translocation. In a mouse model of acute pneumonia, PA103Delta UDelta T reached the lungs as efficiently as PA103Delta U but showed reduced colonization of the liver. This finding suggests that the ability to resist internalization may be important for virulence in vivo.

* Corresponding author. Mailing address: Division of Infectious Disease, Box 0654, University of California, San Francisco, CA 94143-0654. Phone: (415) 476-7355. Fax: (415) 476-9364. E-mail: Jengel{at}medicine.ucsf.edu.

dagger Present address: Department of Bacteriology, University of Wisconsin---Madison, Madison, WI 53706.

Dagger Present address: Department of Microbiology and Immunology, Northwestern University, Chicago, IL 60611.

Infection and Immunity, December 2000, p. 7100-7113, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Endoh, T., Engel, J. N. (2009). CbpA: a Polarly Localized Novel Cyclic AMP-Binding Protein in Pseudomonas aeruginosa. J. Bacteriol. 191: 7193-7205 [Abstract] [Full Text]  
  • 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]  
  • Casaz, P., Garrity-Ryan, L. K., McKenney, D., Jackson, C., Levy, S. B., Tanaka, S. K., Alekshun, M. N. (2006). MarA, SoxS and Rob function as virulence factors in an Escherichia coli murine model of ascending pyelonephritis. Microbiology 152: 3643-3650 [Abstract] [Full Text]  
  • Shafikhani, S. H., Engel, J. (2006). Pseudomonas aeruginosa type III-secreted toxin ExoT inhibits host-cell division by targeting cytokinesis at multiple steps. Proc. Natl. Acad. Sci. USA 103: 15605-15610 [Abstract] [Full Text]  
  • O'Grady, E. P., Mulcahy, H., O'Callaghan, J., Adams, C., O'Gara, F. (2006). Pseudomonas aeruginosa Infection of Airway Epithelial Cells Modulates Expression of Kruppel-Like Factors 2 and 6 via RsmA-Mediated Regulation of Type III Exoenzymes S and Y.. Infect. Immun. 74: 5893-5902 [Abstract] [Full Text]  
  • Zolfaghar, I., Evans, D. J., Ronaghi, R., Fleiszig, S. M. J. (2006). Type III Secretion-Dependent Modulation of Innate Immunity as One of Multiple Factors Regulated by Pseudomonas aeruginosa RetS. Infect. Immun. 74: 3880-3889 [Abstract] [Full Text]  
  • Mulcahy, H., O'Callaghan, J., O'Grady, E. P., Adams, C., O'Gara, F. (2006). The Posttranscriptional Regulator RsmA Plays a Role in the Interaction between Pseudomonas aeruginosa and Human Airway Epithelial Cells by Positively Regulating the Type III Secretion System.. Infect. Immun. 74: 3012-3015 [Abstract] [Full Text]  
  • Shen, D. K., Filopon, D., Kuhn, L., Polack, B., Toussaint, B. (2006). PsrA Is a Positive Transcriptional Regulator of the Type III Secretion System in Pseudomonas aeruginosa. Infect. Immun. 74: 1121-1129 [Abstract] [Full Text]  
  • Shaver, C. M., Hauser, A. R. (2006). Interactions between effector proteins of the Pseudomonas aeruginosa type III secretion system do not significantly affect several measures of disease severity in mammals. Microbiology 152: 143-152 [Abstract] [Full Text]  
  • Deng, Q., Sun, J., Barbieri, J. T. (2005). Uncoupling Crk Signal Transduction by Pseudomonas Exoenzyme T. J. Biol. Chem. 280: 35953-35960 [Abstract] [Full Text]  
  • Kierbel, A., Gassama-Diagne, A., Mostov, K., Engel, J. N. (2005). The Phosphoinositol-3-Kinase-Protein Kinase B/Akt Pathway Is Critical for Pseudomonas aeruginosa Strain PAK Internalization. Mol. Biol. Cell 16: 2577-2585 [Abstract] [Full Text]  
  • Vance, R. E., Rietsch, A., Mekalanos, J. J. (2005). Role of the Type III Secreted Exoenzymes S, T, and Y in Systemic Spread of Pseudomonas aeruginosa PAO1 In Vivo. Infect. Immun. 73: 1706-1713 [Abstract] [Full Text]  
  • Shaver, C. M., Hauser, A. R. (2004). Relative Contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to Virulence in the Lung. Infect. Immun. 72: 6969-6977 [Abstract] [Full Text]  
  • Jain, M., Ramirez, D., Seshadri, R., Cullina, J. F., Powers, C. A., Schulert, G. S., Bar-Meir, M., Sullivan, C. L., McColley, S. A., Hauser, A. R. (2004). Type III Secretion Phenotypes of Pseudomonas aeruginosa Strains Change during Infection of Individuals with Cystic Fibrosis. J. Clin. Microbiol. 42: 5229-5237 [Abstract] [Full Text]  
  • Maresso, A. W., Baldwin, M. R., Barbieri, J. T. (2004). Ezrin/Radixin/Moesin Proteins Are High Affinity Targets for ADP-ribosylation by Pseudomonas aeruginosa ExoS. J. Biol. Chem. 279: 38402-38408 [Abstract] [Full Text]  
  • Goure, J., Pastor, A., Faudry, E., Chabert, J., Dessen, A., Attree, I. (2004). The V Antigen of Pseudomonas aeruginosa Is Required for Assembly of the Functional PopB/PopD Translocation Pore in Host Cell Membranes. Infect. Immun. 72: 4741-4750 [Abstract] [Full Text]  
  • Kazmierczak, B. I., Mostov, K., Engel, J. N. (2004). Epithelial Cell Polarity Alters Rho-GTPase Responses to Pseudomonas aeruginosa. Mol. Biol. Cell 15: 411-419 [Abstract] [Full Text]  
  • Schmidt, H., Hensel, M. (2004). Pathogenicity Islands in Bacterial Pathogenesis. Clin. Microbiol. Rev. 17: 14-56 [Abstract] [Full Text]  
  • Garrity-Ryan, L., Shafikhani, S., Balachandran, P., Nguyen, L., Oza, J., Jakobsen, T., Sargent, J., Fang, X., Cordwell, S., Matthay, M. A., Engel, J. N. (2004). The ADP Ribosyltransferase Domain of Pseudomonas aeruginosa ExoT Contributes to Its Biological Activities. Infect. Immun. 72: 546-558 [Abstract] [Full Text]  
  • Phillips, R. M., Six, D. A., Dennis, E. A., Ghosh, P. (2003). In Vivo Phospholipase Activity of the Pseudomonas aeruginosa Cytotoxin ExoU and Protection of Mammalian Cells with Phospholipase A2 Inhibitors. J. Biol. Chem. 278: 41326-41332 [Abstract] [Full Text]  
  • McMorran, B., Town, L., Costelloe, E., Palmer, J., Engel, J., Hume, D., Wainwright, B. (2003). Effector ExoU from the Type III Secretion System Is an Important Modulator of Gene Expression in Lung Epithelial Cells in Response to Pseudomonas aeruginosa Infection. Infect. Immun. 71: 6035-6044 [Abstract] [Full Text]  
  • Lee, E. J., Cowell, B. A., Evans, D. J., Fleiszig, S. M. J. (2003). Contribution of ExsA-Regulated Factors to Corneal Infection by Cytotoxic and Invasive Pseudomonas aeruginosa in a Murine Scarification Model. IOVS 44: 3892-3898 [Abstract] [Full Text]  
  • Sun, J., Barbieri, J. T. (2003). Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 Regulator of Kinase (Crk) Proteins. J. Biol. Chem. 278: 32794-32800 [Abstract] [Full Text]  
  • Ajayi, T., Allmond, L. R., Sawa, T., Wiener-Kronish, J. P. (2003). Single-Nucleotide-Polymorphism Mapping of the Pseudomonas aeruginosa Type III Secretion Toxins for Development of a Diagnostic Multiplex PCR System. J. Clin. Microbiol. 41: 3526-3531 [Abstract] [Full Text]  
  • Cowell, B. A., Twining, S. S., Hobden, J. A., Kwong, M. S. F., Fleiszig, S. M. J. (2003). Mutation of lasA and lasB reduces Pseudomonas aeruginosa invasion of epithelial cells. Microbiology 149: 2291-2299 [Abstract] [Full Text]  
  • Miyata, S., Casey, M., Frank, D. W., Ausubel, F. M., Drenkard, E. (2003). Use of the Galleria mellonella Caterpillar as a Model Host To Study the Role of the Type III Secretion System in Pseudomonas aeruginosa Pathogenesis. Infect. Immun. 71: 2404-2413 [Abstract] [Full Text]  
  • Jendrossek, V., Fillon, S., Belka, C., Muller, I., Puttkammer, B., Lang, F. (2003). Apoptotic Response of Chang Cells to Infection with Pseudomonas aeruginosa Strains PAK and PAO-I: Molecular Ordering of the Apoptosis Signaling Cascade and Role of Type IV Pili. Infect. Immun. 71: 2665-2673 [Abstract] [Full Text]  
  • Jacob, T., Lee, R. J., Engel, J. N., Machen, T. E. (2002). Modulation of Cytosolic Ca2+ Concentration in Airway Epithelial Cells by Pseudomonas aeruginosa. Infect. Immun. 70: 6399-6408 [Abstract] [Full Text]  
  • Kazmierczak, B. I., Engel, J. N. (2002). Pseudomonas aeruginosa ExoT Acts In Vivo as a GTPase-Activating Protein for RhoA, Rac1, and Cdc42. Infect. Immun. 70: 2198-2205 [Abstract] [Full Text]  
  • Krall, R., Sun, J., Pederson, K. J., Barbieri, J. T. (2002). In Vivo Rho GTPase-Activating Protein Activity of Pseudomonas aeruginosa Cytotoxin ExoS. Infect. Immun. 70: 360-367 [Abstract] [Full Text]  
  • Feltman, H., Schulert, G., Khan, S., Jain, M., Peterson, L., Hauser, A. R. (2001). Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa. Microbiology 147: 2659-2669 [Abstract] [Full Text]  
  • Fraylick, J. E., La Rocque, J. R., Vincent, T. S., Olson, J. C. (2001). Independent and Coordinate Effects of ADP-Ribosyltransferase and GTPase-Activating Activities of Exoenzyme S on HT-29 Epithelial Cell Function. Infect. Immun. 69: 5318-5328 [Abstract] [Full Text]  
  • Ferguson, M. W., Maxwell, J. A., Vincent, T. S., da Silva, J., Olson, J. C. (2001). Comparison of the exoS Gene and Protein Expression in Soil and Clinical Isolates of Pseudomonas aeruginosa. Infect. Immun. 69: 2198-2210 [Abstract] [Full Text]  


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