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Journal of Bacteriology, June 2006, p. 3995-4006, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00053-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Contribution of the PhoP-PhoQ and PmrA-PmrB Two-Component Regulatory Systems to Mg2+-Induced Gene Regulation in Pseudomonas aeruginosa

Joseph B. McPhee,1 Manjeet Bains,1 Geoff Winsor,2 Shawn Lewenza,1 Agnieszka Kwasnicka,1 Michelle D. Brazas,1 Fiona S. L. Brinkman,2 and R. E. W. Hancock1*

Department of Microbiology and Immunology and Centre for Microbial Diseases and Immunity Research, University of British Columbia, 232-2259 Lower Mall, Vancouver, BC, Canada V6T 1Z4,1 Department of Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Dr., Burnaby, BC, Canada V5A 1S622

Received 12 January 2006/ Accepted 10 March 2006

When grown in divalent cation-limited medium, Pseudomonas aeruginosa becomes resistant to cationic antimicrobial peptides and polymyxin B. This resistance is regulated by the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems. To further characterize Mg2+ regulation in P. aeruginosa, microarray transcriptional profiling was conducted to compare wild-type P. aeruginosa grown under Mg2+-limited and Mg2+-replete conditions to isogenic phoP and pmrA mutants grown under Mg2+-limited conditions. Under Mg2+-limited conditions (0.02 mM Mg2+), approximately 3% of the P. aeruginosa genes were differentially expressed compared to the expression in bacteria grown under Mg2+-replete conditions (2 mM Mg2+). Only a modest subset of the Mg2+-regulated genes were regulated through either PhoP or PmrA. To determine which genes were directly regulated, a bioinformatic search for conserved binding motifs was combined with confirmatory reverse transcriptase PCR and gel shift promoter binding assays, and the results indicated that very few genes were directly regulated by these response regulators. It was found that in addition to the previously known oprH-phoP-phoQ operon and the pmrHFIJKLM-ugd operon, the PA0921 and PA1343 genes, encoding small basic proteins, were regulated by Mg2+ in a PhoP-dependent manner. The number of known PmrA-regulated genes was expanded to include the PA1559-PA1560, PA4782-PA4781, and feoAB operons, in addition to the previously known PA4773-PA4775-pmrAB and pmrHFIJKLM-ugd operons.

* Corresponding author. Mailing address: Department of Microbiology and Immunology and Centre for Microbial Diseases and Immunity Research, University of British Columbia, 232-2259 Lower Mall, Vancouver, BC, Canada V6T 1Z4. Phone: (604) 822-2682. Fax: (604) 827-5566. E-mail: bob{at}cmdr.ubc.ca.

Journal of Bacteriology, June 2006, p. 3995-4006, Vol. 188, No. 11
0021-9193/06/$08.00+0     doi:10.1128/JB.00053-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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