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Journal of Bacteriology, October 1999, p. 6300-6305, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Characterization of MexT, the Regulator of the MexE-MexF-OprN
Multidrug Efflux System of Pseudomonas aeruginosa
Thilo
Köhler,*
Simone F.
Epp,
Lasta Kocjancic
Curty, and
Jean-Claude
Pechère
Department of Genetics and Microbiology,
Centre Médical Universitaire, Geneva, Switzerland
Received 20 April 1999/Accepted 5 August 1999
We investigated the regulation of the MexEF-OprN multidrug efflux
system of Pseudomonas aeruginosa, which is overexpressed in
nfxC-type mutants and confers resistance to quinolones,
chloramphenicol and trimethoprim. Sequencing of the DNA region upstream
of the mexEF-oprN operon revealed the presence of an open
reading frame (ORF) of 304 amino acids encoding a LysR-type
transcriptional activator, termed MexT. By using T7-polymerase, a
34-kDa protein was expressed in Escherichia coli from a
plasmid carrying the mexT gene. Expression of a
mexE::lacZ fusion was 10-fold higher in nfxC-type mutants than in the wild-type strain; however,
transcription of mexT as well as the mexT DNA
region was unchanged. Located adjacent to mexT but
transcribed in opposite direction, the beginning of an ORF termed
qrh (quinone oxidoreductase homologue) was identified. Expression of a qrh::lacZ fusion was
also found to be activated by MexT. Further, we present evidence for
coregulation at the transcriptional and the posttranscriptional level
between the MexEF-OprN efflux system and the OprD porin responsible for
cross-resistance of nfxC-type mutants to carbapenem antibiotics.
*
Corresponding author. Mailing address: Department of
Genetics and Microbiology, CMU 1, rue Michel Servet, CH-1211 Geneva 4, Switzerland. Phone: 41-22-7025655. Fax: 41-22-7025702. E-mail: Thilo.Kohler{at}medecine.unige.ch.
Journal of Bacteriology, October 1999, p. 6300-6305, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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