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Antimicrobial Agents and Chemotherapy, January 2003, p. 95-101, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.95-101.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Regulation of Membrane Permeability by a Two-Component Regulatory System in Pseudomonas aeruginosa

Yanping Wang, Unhwan Ha, Lin Zeng, and Shouguang Jin*

Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida 32610

Received 7 June 2002/ Returned for modification 19 August 2002/ Accepted 10 October 2002

Membrane impermeability is the major contributing factor to multidrug resistance in clinical isolates of Pseudomonas aeruginosa. By using laboratory strain PAK, a spontaneous P. aeruginosa mutant (mutant PAK1-3) whose membrane had reduced permeability and which displayed increased levels of resistance to various antibiotics, especially aminoglycosides, was isolated. By complementation of the mutant with a genomic clone library derived from wild-type strain PAK, a novel two-component regulatory system (PprA and PprB) was identified and was found to be able to increase the permeability of the bacterial membrane and render PAK1-3 sensitive to antibiotics. Furthermore, specific phosphorylation of the response regulator (PprB) by histidine kinase (PprA) was observed in vitro, demonstrating that they are cognate two-component regulatory genes. Introduction of a plasmid expressing the pprB gene into randomly chosen clinical isolates (n = 17) resulted in increased sensitivity to aminoglycosides in the majority of isolates (n = 13) tested. This is the first demonstration that P. aeruginosa membrane permeability can be regulated, providing an important clue in the understanding of the mechanism of membrane impermeability-mediated multidrug resistance in P. aeruginosa.

* Corresponding author. Mailing address: Department of Molecular Genetics and Microbiology, P.O. Box 100266, University of Florida, Gainesville, FL 32610. Phone: (352) 392-8323. Fax: (352) 392-3133. E-mail: sjin{at}mgm.ufl.edu.

Antimicrobial Agents and Chemotherapy, January 2003, p. 95-101, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.95-101.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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