Abstract

The primary goal of the proposed investigation is a detailed analysis of the cellular and molecular aspects of localization of two pseudomonas aeruginosa virulence factors: exotoxin A and the pilus adhesin. Biochemical and genetic tools will be used to identify the pathway for the export of these proteins. The export pathway will be studied by examining the various steps of cytoplasmic membrane traversal and translocation across the outer membrane. Techniques of protein fusion and site directed mutagenesis will be used to identify domains in exotoxin A and pilin, that play a role in various steps of membrane translocation. Mutants of P. aeruginosa defective in export of exotoxin A and pilin will be identified and characterized. Defective genes in such mutants will be complemented by recombinant plasmids, and coding sequences for the export genes will be isolated from such plasmids. These studies should not only help in a better understanding of regulation of biosynthesis of virulence factors by P. aeruginosa, but also open new lines of investigation into the mechanism of extracellular protein localization by Gram-negative bacteria.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI021451-08
Application #
3131589
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Project Start
1984-08-01
Project End
1994-03-31
Budget Start
1993-02-01
Budget End
1994-03-31
Support Year
8
Fiscal Year
1993
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Skurnik, David; Roux, Damien; Aschard, Hugues et al. (2013) A comprehensive analysis of in vitro and in vivo genetic fitness of Pseudomonas aeruginosa using high-throughput sequencing of transposon libraries. PLoS Pathog 9:e1003582
Koh, Andrew Y; Mikkelsen, Per J; Smith, Roger S et al. (2010) Utility of in vivo transcription profiling for identifying Pseudomonas aeruginosa genes needed for gastrointestinal colonization and dissemination. PLoS One 5:e15131
Thaden, Joshua T; Lory, Stephen; Gardner, Timothy S (2010) Quorum-sensing regulation of a copper toxicity system in Pseudomonas aeruginosa. J Bacteriol 192:2557-68
Mulcahy, Lawrence R; Burns, Jane L; Lory, Stephen et al. (2010) Emergence of Pseudomonas aeruginosa strains producing high levels of persister cells in patients with cystic fibrosis. J Bacteriol 192:6191-9
Hurley, Bryan P; Goodman, Andrew L; Mumy, Karen L et al. (2010) The two-component sensor response regulator RoxS/RoxR plays a role in Pseudomonas aeruginosa interactions with airway epithelial cells. Microbes Infect 12:190-8
Lory, Stephen; Merighi, Massimo; Hyodo, Mamoru (2009) Multiple activities of c-di-GMP in Pseudomonas aeruginosa. Nucleic Acids Symp Ser (Oxf) :51-2
Goodman, Andrew L; Merighi, Massimo; Hyodo, Mamoru et al. (2009) Direct interaction between sensor kinase proteins mediates acute and chronic disease phenotypes in a bacterial pathogen. Genes Dev 23:249-59
Brencic, Anja; McFarland, Kirsty A; McManus, Heather R et al. (2009) The GacS/GacA signal transduction system of Pseudomonas aeruginosa acts exclusively through its control over the transcription of the RsmY and RsmZ regulatory small RNAs. Mol Microbiol 73:434-45
Brencic, Anja; Lory, Stephen (2009) Determination of the regulon and identification of novel mRNA targets of Pseudomonas aeruginosa RsmA. Mol Microbiol 72:612-32
Mougous, Joseph D; Cuff, Marianne E; Raunser, Stefan et al. (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science 312:1526-30

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