Identification of Pseudomonas aeruginosa genes required for epithelial cell injury

Pil Jung Kang, Alan R. Hauser, Gerard Apodaca, Suzanne M J Fleiszig, Jeanine Wiener-Kronish, Keith Mostov, Joanne N. Engel*

*Corresponding author for this work

Research output: Contribution to journalArticle

105 Scopus citations

Abstract

We have developed a simple, reproducible and rapid genetic screen for Pseudomonas aeruginosa-induced epithelial cell cytotoxicity in cultures of MDCK cells. This screen was used to isolate isogenic transposon-tagged non- cytotoxic mutants of a cytotoxic and lung-virulent strain of P. aeruginosa (PA103). The transposon-insertion site was determined by using an inverse polymerase chain reaction followed by DNA-sequence analysis. On the basis of phenotype and sequence analysis, these mutants fell into four classes. One class had absent or defective pili, based on their resistance to phage PO4 and/or loss of twitching motility (twt). A second class exhibited decreased adherence. A third class of mutants exhibited probable defects in the machinery or targets of type III protein secretion. A final class of mutants exhibited decreased but not absent cytotoxicity. This class included members of the first three classes as well as other mutants. These results suggest that localized cytotoxicity is likely to require several steps and several components, including pill and other (unidentified) extracellular proteins. The type III protein-secretion apparatus appears to be involved in this process.

Original languageEnglish (US)
Pages (from-to)1249-1262
Number of pages14
JournalMolecular Microbiology
Volume24
Issue number6
StatePublished - Jul 24 1997

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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    Kang, P. J., Hauser, A. R., Apodaca, G., Fleiszig, S. M. J., Wiener-Kronish, J., Mostov, K., & Engel, J. N. (1997). Identification of Pseudomonas aeruginosa genes required for epithelial cell injury. Molecular Microbiology, 24(6), 1249-1262.