This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Olson, J. C. Articles by Frank, D. W. Search for Related Content PubMed PubMed Citation Articles by Olson, J. C. Articles by Frank, D. W.

 Previous Article  |  Next Article 

Infect. Immun., 01 1997, 248-256, Vol 65, No. 1
Copyright © 1997, American Society for Microbiology

Effects of differential expression of the 49-kilodalton exoenzyme S by Pseudomonas aeruginosa on cultured eukaryotic cells

JC Olson, EM McGuffie and DW Frank
Department of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston 29425, USA. olsonjc@musc.edu

Production of the ADP-ribosylating enzyme exoenzyme S (ExoS) by Pseudomonas aeruginosa has been associated with increased virulence. Previous studies, however, have been unable to confirm an effect of soluble ExoS in cell culture or animal model systems. To determine if bacteria must come in contact with target cells in order for an effect of ExoS to be observed, coculture systems were developed to compare the effects of ExoS- and non-ExoS-producing bacteria on eukaryotic cell function. The two P. aeruginosa strains used in these studies, 388 and 388delta exoS, maintained genetic identity, with the exception that strain 388delta exoS lacked production of the 49-kDa form of ExoS. When bacteria were cocultured with Detroit 532 fibroblastic cells, ExoS- producing 388 bacteria caused a significant decrease in DNA synthesis and viability compared to the decrease caused by non-ExoS-producing 388delta exoS bacteria. Maximal differences between the two strains were observed when 10(4) to 10(7) CFU of bacteria/ml were cocultured with Detroit cells for 4 or 6 h. Both strains were effective in eliminating Detroit cell DNA synthesis after a 20-h coculture period. Secreted ExoS had no effect on Detroit cell growth and viability, indicating that bacteria must have contact with target cells for the effect of ExoS on cellular function to be observed. Similar effects on cell proliferation and viability were observed when the two strains were cocultured with the KB epithelioid cell line. ExoS-associated decreases in eukaryotic cell viability were not found to be mediated by an inhibition of protein synthesis. These studies confirm that the 49- kDa ExoS contributes to the cellular pathogenesis of P. aeruginosa by interfering with eukaryotic cell growth and viability. In addition, the coculture system developed which recognizes this effect should provide a means for defining the function of ExoS in vivo.

This article has been cited by other articles:

• 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]  
• 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]  
• Rocha, C. L., Rucks, E. A., Vincent, D. M., Olson, J. C. (2005). Examination of the Coordinate Effects of Pseudomonas aeruginosa ExoS on Rac1. Infect. Immun. 73: 5458-5467 [Abstract] [Full Text]  
• Rucks, E. A., Olson, J. C. (2005). Characterization of an ExoS Type III Translocation-Resistant Cell Line. Infect. Immun. 73: 638-643 [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]  
• Rocha, C. L., Coburn, J., Rucks, E. A., Olson, J. C. (2003). Characterization of Pseudomonas aeruginosa Exoenzyme S as a Bifunctional Enzyme in J774A.1 Macrophages. Infect. Immun. 71: 5296-5305 [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]  
• Rucks, E. A., Fraylick, J. E., Brandt, L. M., Vincent, T. S., Olson, J. C. (2003). Cell line differences in bacterially translocated ExoS ADP-ribosyltransferase substrate specificity. Microbiology 149: 319-331 [Abstract] [Full Text]  
• Riese, M. J., Goehring, U.-M., Ehrmantraut, M. E., Moss, J., Barbieri, J. T., Aktories, K., Schmidt, G. (2002). Auto-ADP-ribosylation of Pseudomonas aeruginosa ExoS. J. Biol. Chem. 277: 12082-12088 [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]  
• Dacheux, D., Attree, I., Toussaint, B. (2001). Expression of ExsA in trans Confers Type III Secretion System-Dependent Cytotoxicity on Noncytotoxic Pseudomonas aeruginosa Cystic Fibrosis Isolates. Infect. Immun. 69: 538-542 [Abstract] [Full Text]  
• Dacheux, D., Toussaint, B., Richard, M., Brochier, G., Croize, J., Attree, I. (2000). Pseudomonas aeruginosa Cystic Fibrosis Isolates Induce Rapid, Type III Secretion-Dependent, but ExoU-Independent, Oncosis of Macrophages and Polymorphonuclear Neutrophils. Infect. Immun. 68: 2916-2924 [Abstract] [Full Text]  
• Dacheux, D., Attree, I., Schneider, C., Toussaint, B. (1999). Cell Death of Human Polymorphonuclear Neutrophils Induced by a Pseudomonas aeruginosa Cystic Fibrosis Isolate Requires a Functional Type III Secretion System. Infect. Immun. 67: 6164-6167 [Abstract] [Full Text]  
• Hauser, A. R., Engel, J. N. (1999). Pseudomonas aeruginosa Induces Type-III-Secretion-Mediated Apoptosis of Macrophages and Epithelial Cells. Infect. Immun. 67: 5530-5537 [Abstract] [Full Text]  
• Ganesan, A. K., Vincent, T. S., Olson, J. C., Barbieri, J. T. (1999). Pseudomonas aeruginosa Exoenzyme S Disrupts Ras-mediated Signal Transduction by Inhibiting Guanine Nucleotide Exchange Factor-catalyzed Nucleotide Exchange. J. Biol. Chem. 274: 21823-21829 [Abstract] [Full Text]  
• Darwin, K. H., Miller, V. L. (1999). Molecular Basis of the Interaction of Salmonella with the Intestinal Mucosa. Clin. Microbiol. Rev. 12: 405-428 [Abstract] [Full Text]  
• McGuffie, E. M., Fraylick, J. E., Hazen-Martin, D. J., Vincent, T. S., Olson, J. C. (1999). Differential Sensitivity of Human Epithelial Cells to Pseudomonas aeruginosa Exoenzyme S. Infect. Immun. 67: 3494-3503 [Abstract] [Full Text]  
• Olson, J. C., Fraylick, J. E., McGuffie, E. M., Dolan, K. M., Yahr, T. L., Frank, D. W., Vincent, T. S. (1999). Interruption of Multiple Cellular Processes in HT-29 Epithelial Cells by Pseudomonas aeruginosa Exoenzyme S. Infect. Immun. 67: 2847-2854 [Abstract] [Full Text]  
• Coburn, J., Frank, D. W. (1999). Macrophages and Epithelial Cells Respond Differently to the Pseudomonas aeruginosa Type III Secretion System. Infect. Immun. 67: 3151-3154 [Abstract] [Full Text]  
• Ganesan, A. K., Mende-Mueller, L., Selzer, J., Barbieri, J. T. (1999). Pseudomonas aeruginosa Exoenzyme S, a Double ADP-ribosyltransferase, Resembles Vertebrate Mono-ADP-ribosyltransferases. J. Biol. Chem. 274: 9503-9508 [Abstract] [Full Text]  
• Vallis, A. J., Finck-Barbancon, V., Yahr, T. L., Frank, D. W. (1999). Biological Effects of Pseudomonas aeruginosa Type III-Secreted Proteins on CHO Cells. Infect. Immun. 67: 2040-2044 [Abstract] [Full Text]  
• Vallis, A. J., Yahr, T. L., Barbieri, J. T., Frank, D. W. (1999). Regulation of ExoS Production and Secretion by Pseudomonas aeruginosa in Response to Tissue Culture Conditions. Infect. Immun. 67: 914-920 [Abstract] [Full Text]  
• Finck-Barbançon, V., Yahr, T. L., Frank, D. W. (1998). Identification and Characterization of SpcU, a Chaperone Required for Efficient Secretion of the ExoU Cytotoxin. J. Bacteriol. 180: 6224-6231 [Abstract] [Full Text]  
• Bruno, T. F., Buser, D. E., Syme, R. M., Woods, D. E., Mody, C. H. (1998). Pseudomonas aeruginosa Exoenzyme S Is a Mitogen but Not a Superantigen for Human T Lymphocytes. Infect. Immun. 66: 3072-3079 [Abstract] [Full Text]  
• McGuffie, E. M., Frank, D. W., Vincent, T. S., Olson, J. C. (1998). Modification of Ras in Eukaryotic Cells by Pseudomonas aeruginosa Exoenzyme S. Infect. Immun. 66: 2607-2613 [Abstract] [Full Text]