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Mechanisms of Action: Physiological Effects

Ciprofloxacin Induction of a Susceptibility Determinant in Pseudomonas aeruginosa

Michelle D. Brazas, Robert E. W. Hancock
Michelle D. Brazas
Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada
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Robert E. W. Hancock
Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada
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  • For correspondence: bob@cmdr.ubc.ca
DOI: 10.1128/AAC.49.8.3222-3227.2005
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ABSTRACT

With few novel antimicrobials in development, resistance to the current selection of antibiotics increasingly encroaches on our ability to control microbial infections. One limitation in our understanding of the basis of the constraints on current therapies is our poor understanding of antibiotic interactions with bacteria on a global scale. Custom DNA microarrays were used to characterize the response of Pseudomonas aeruginosa to ciprofloxacin, a fluoroquinolone commonly used in therapy against chronic infections by this intrinsically resistant bacterium. Of the approximately 5,300 open reading frames (ORFs) on the array, 941 genes showed statistically significant (P ≤ 0.05) differential expression in response to 0.3× MIC of ciprofloxacin; 554 were promoted and 387 were repressed. Most striking among the responsive genes was the region between PA0613 and PA0648, which codes for the bacteriophage-like R2/F2 pyocins. In this region, virtually every ORF was increased by 0.3× MIC of ciprofloxacin and even more dramatically up-regulated (7- to 19-fold) following treatment with 1× MIC of ciprofloxacin. Pyocin gene expression was confirmed with lux reporter mutants and real-time PCR studies; pyocin-like particles were also present in transmission electron micrographs of supernatants from cells treated with 1× MIC of ciprofloxacin. Interestingly, mutants in this region exhibited ≥8-fold-increased resistance to ciprofloxacin and other fluoroquinolones, demonstrating that this region is a susceptibility determinant. Since this region is known to be variably present in the genomes of clinical isolates of P. aeruginosa (R. K. Ernst et al., Environ. Microbiol. 5:1341-1349, 2003, and M. C. Wolfgang et al., Proc. Natl. Acad. Sci. USA 100:8484-8489, 2003), these findings demonstrate that the R2/F2 pyocin region is a “loaded gun” that can mediate fluoroquinolone susceptibility in P. aeruginosa.

  • Copyright © 2005 American Society for Microbiology
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Ciprofloxacin Induction of a Susceptibility Determinant in Pseudomonas aeruginosa
Michelle D. Brazas, Robert E. W. Hancock
Antimicrobial Agents and Chemotherapy Jul 2005, 49 (8) 3222-3227; DOI: 10.1128/AAC.49.8.3222-3227.2005

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Ciprofloxacin Induction of a Susceptibility Determinant in Pseudomonas aeruginosa
Michelle D. Brazas, Robert E. W. Hancock
Antimicrobial Agents and Chemotherapy Jul 2005, 49 (8) 3222-3227; DOI: 10.1128/AAC.49.8.3222-3227.2005
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KEYWORDS

anti-infective agents
ciprofloxacin
Gene Expression Regulation, Bacterial
Pseudomonas aeruginosa

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