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Antimicrobial Agents and Chemotherapy, March 2004, p. 843-851, Vol. 48, No. 3
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.3.843-851.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Selection of an Antibiotic-Hypersusceptible Mutant of Pseudomonas aeruginosa: Identification of the GlmR Transcriptional Regulator

Julio Ramos-Aires,1,2 Patrick Plésiat,3 Lasta Kocjancic-Curty,1 and Thilo Köhler1*

Department of Genetics and Microbiology, University of Geneva, CH-1211 Geneva 4, Switzerland,1 Laboratoire de Bactériologie, Hôpital J. Minjoz, Besançon, France,3 Department of Molecular Biology, University of California, Berkeley, California2

Received 30 May 2003/ Returned for modification 18 August 2003/ Accepted 26 November 2003

Tn501 random mutagenesis was applied to the Pseudomonas aeruginosa wild-type strain PAO1 to select for mutants hypersusceptible to aminoglycoside antimicrobial agents. One such mutant, called 19A, was found to be hypersusceptible to a wide range of antibiotics including aminoglycosides, ß-lactams, fluoroquinolones, colistin, erythromycin, rifampin, and glycopeptides. Light microscopy of the mutant strain revealed abnormal morphology characterized by large, filamentous cells. The drug supersusceptibility of 19A was accompanied by loss of motility, reduced resistance to osmotic and heat shock stress, and impaired growth at low temperatures. The insertion site of the Tn501 transposon in mutant 19A has occurred in an open reading frame (PA5550 according to the PAO1 genome project), whose gene product shows amino acid sequence similarity to the DeoR family of transcriptional repressors. The gene, which we called glmR, is located between the glmS (PA5549) and glmU (PA5552) homologues of E. coli, responsible for the synthesis of UDP-N-acetylglucosamine-1-P, a precursor of both lipopolysaccharide (LPS) and peptidoglycan. We showed that GlmR represses the transcription of the adjacent glmS homologue (PA5549) in P. aeruginosa, possibly affecting the pool of precursors for peptidoglycan and LPS synthesis. To our knowledge GlmR is the first regulator in P. aeruginosa that affects susceptibility to a large variety of antibiotics and is therefore a potential target for novel anti-infective agents.

* Corresponding author. Mailing address: Department of Genetics and Microbiology, University of Geneva, 1 rue Michel Servet, CH-1211 Geneva 4, Switzerland. Phone: (41) 22 379-56-55. Fax: (41) 22 379-57-02. E-mail: Thilo.Kohler{at}medecine.unige.ch.

Antimicrobial Agents and Chemotherapy, March 2004, p. 843-851, Vol. 48, No. 3
0066-4804/04/$08.00+0     DOI: 10.1128/AAC.48.3.843-851.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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