This Article
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 Yahr, T. L.
Right arrow Articles by Frank, D. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yahr, T. L.
Right arrow Articles by Frank, D. W.

 Previous Article  |  Next Article 

J. Bacteriol., 03 1996, 1412-1419, Vol 178, No. 5
Copyright © 1996, American Society for Microbiology

Genetic relationship between the 53- and 49-kilodalton forms of exoenzyme S from Pseudomonas aeruginosa

TL Yahr, JT Barbieri and DW Frank
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226, USA.

Exoenzyme S is an ADP-ribosylating extracellular protein of Pseudomonas aeruginosa that is produced as two immunologically related forms, a 49- kDa enzymatically active form and a 53-kDa inactive form. The postulated relationship between the two proteins involves a carboxy- terminal proteolytic cleavage of the 53-kDa precursor to produce an enzymatically active 49-kDa protein. To determine the genetic relationship between the two forms of exoenzyme S, exoS (encoding the 49-kDa form) was used as a probe in Southern blot analyses of P. aeruginosa chromosomal digests. Cross-hybridizing bands were detected in chromosomal digests of a strain of P. aeruginosa in which exoS had been deleted by allelic exchange. A chromosomal bank was prepared from the exoS deletion strain, 388deltaexoS::TC, and screened with a probe internal to exoS. Thirteen clones that cross-hybridized with the exoS probe were identified. One representative clone contained the open reading frame exoT; this open reading frame encoded a protein of 457 amino acids which showed 75% amino acid identity to ExoS. The exoT open reading frame, cloned into a T7 expression system, produced a 53-kDa protein in Escherichia coli, termed Exo53, which reacted to antisera against exoenzyme S. A histidine-tagged derivative of recombinant Exo53 possessed approximately 0.2% of the ADP-ribosyltransferase activity of recombinant ExoS. Inactivation of exoT in an allelic-replacement strain resulted in an Exo53-deficient phenotype without modifying the expression of ExoS. These studies prove that the 53- and 49-kDa forms of exoenzyme S are encoded by separate genes. In addition, this is the first report of the factor-activating-exoenzyme-S-dependent ADP- ribosyltransferase activity of the 53-kDa form of exoenzyme S.


This article has been cited by other articles:

  • Imamura, Y., Yanagihara, K., Fukuda, Y., Kaneko, Y., Seki, M., Izumikawa, K., Miyazaki, Y., Hirakata, Y., Sawa, T., Wiener-Kronish, J. P., Kohno, S. (2007). Effect of anti-PcrV antibody in a murine chronic airway Pseudomonas aeruginosa infection model. Eur Respir J 29: 965-968 [Abstract] [Full Text]  
  • Rietsch, A., Vallet-Gely, I., Dove, S. L., Mekalanos, J. J. (2005). ExsE, a secreted regulator of type III secretion genes in Pseudomonas aeruginosa. Proc. Natl. Acad. Sci. USA 102: 8006-8011 [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]  
  • Schaber, J. A., Carty, N. L., McDonald, N. A., Graham, E. D., Cheluvappa, R., Griswold, J. A., Hamood, A. N. (2004). Analysis of quorum sensing-deficient clinical isolates of Pseudomonas aeruginosa. J Med Microbiol 53: 841-853 [Abstract] [Full Text]  
  • Rietsch, A., Wolfgang, M. C., Mekalanos, J. J. (2004). Effect of Metabolic Imbalance on Expression of Type III Secretion Genes in Pseudomonas aeruginosa. Infect. Immun. 72: 1383-1390 [Abstract] [Full Text]  
  • Smith, R. S., Wolfgang, M. C., Lory, S. (2004). An Adenylate Cyclase-Controlled Signaling Network Regulates Pseudomonas aeruginosa Virulence in a Mouse Model of Acute Pneumonia. Infect. Immun. 72: 1677-1684 [Abstract] [Full Text]  
  • Ahn, K.-S., Ha, U., Jia, J., Wu, D., Jin, S. (2004). The truA gene of Pseudomonas aeruginosa is required for the expression of type III secretory genes. Microbiology 150: 539-547 [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]  
  • 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]  
  • Jia, J., Alaoui-El-Azher, M., Chow, M., Chambers, T. C., Baker, H., Jin, S. (2003). c-Jun NH2-Terminal Kinase-Mediated Signaling Is Essential for Pseudomonas aeruginosa ExoS-Induced Apoptosis. Infect. Immun. 71: 3361-3370 [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]  
  • 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]  
  • Braun, M., Stuber, K., Schlatter, Y., Wahli, T., Kuhnert, P., Frey, J. (2002). Characterization of an ADP-Ribosyltransferase Toxin (AexT) from Aeromonas salmonicida subsp. salmonicida. J. Bacteriol. 184: 1851-1858 [Abstract] [Full Text]  
  • Lomholt, J. A., Poulsen, K., Kilian, M. (2001). Epidemic Population Structure of Pseudomonas aeruginosa: Evidence for a Clone That Is Pathogenic to the Eye and That Has a Distinct Combination of Virulence Factors. Infect. Immun. 69: 6284-6295 [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]  
  • Holder, I. A., Neely, A. N., Frank, D. W. (2001). PcrV Immunization Enhances Survival of Burned Pseudomonas aeruginosa-Infected Mice. Infect. Immun. 69: 5908-5910 [Abstract] [Full Text]  
  • Ha, U., Jin, S. (2001). Growth Phase-Dependent Invasion of Pseudomonas aeruginosa and Its Survival within HeLa Cells. Infect. Immun. 69: 4398-4406 [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]  
  • Garrity-Ryan, L., Kazmierczak, B., Kowal, R., Comolli, J., Hauser, A., Engel, J. N. (2000). The Arginine Finger Domain of ExoT Contributes to Actin Cytoskeleton Disruption and Inhibition of Internalization of Pseudomonas aeruginosa by Epithelial Cells and Macrophages. Infect. Immun. 68: 7100-7113 [Abstract] [Full Text]  
  • Krall, R., Schmidt, G., Aktories, K., Barbieri, J. T. (2000). Pseudomonas aeruginosa ExoT Is a Rho GTPase-Activating Protein. Infect. Immun. 68: 6066-6068 [Abstract] [Full Text]  
  • Kaufman, M. R., Jia, J., Zeng, L., Ha, U., Chow, M., Jin, S. (2000). Pseudomonas aeruginosa mediated apoptosis requires the ADP-ribosylating activity of ExoS. Microbiology 146: 2531-2541 [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]  
  • Cowell, B. A., Chen, D. Y., Frank, D. W., Vallis, A. J., Fleiszig, S. M. J. (2000). ExoT of Cytotoxic Pseudomonas aeruginosa Prevents Uptake by Corneal Epithelial Cells. Infect. Immun. 68: 403-406 [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]  
  • 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]  
  • 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]  
  • Radke, J., Pederson, K. J., Barbieri, J. T. (1999). Pseudomonas aeruginosa Exoenzyme S Is a Biglutamic Acid ADP-Ribosyltransferase. Infect. Immun. 67: 1508-1510 [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]  
  • Yahr, T. L., Vallis, A. J., Hancock, M. K., Barbieri, J. T., Frank, D. W. (1998). ExoY, an adenylate cyclase secreted by the Pseudomonas aeruginosa type III system. Proc. Natl. Acad. Sci. USA 95: 13899-13904 [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]  


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