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Antimicrobial Agents and Chemotherapy, July 1998, p. 1682-1688, Vol. 42, No. 7
Department of Microbiology and Immunology,
Queen's University, Kingston, Ontario K7L 3N6, Canada
Received 23 February 1998/Returned for modification 16 April
1998/Accepted 6 May 1998
A Pseudomonas aeruginosa strain carrying an insertion
of an
0066-4804/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Contribution of Outer Membrane Efflux Protein OprM
to Antibiotic Resistance in Pseudomonas
aeruginosa Independent of MexAB
Hg interposon in the mexB gene
(mexB::
Hg; strain K879) produced markedly
reduced but still detectable levels of OprM, the product of the third
gene of the mexAB-oprM multidrug efflux operon. By using a
lacZ transcriptional fusion vector, promoter activity likely responsible for OprM expression in the
mexB::
Hg mutant was identified upstream of
oprM. Introduction of the oprM gene, but not
the mexAB genes, into a P. aeruginosa
multidrug-susceptible
mexAB-oprM mutant
increased resistance to quinolones, cephalosporins, erythromycin,
and tetracycline. A
mexAB-oprM strain
carrying the oprM gene accumulated markedly less
antibiotic than the deletion strain without oprM.
Antibiotic accumulation by the MexAB
OprM+
strain was markedly enhanced upon treatment of cells with the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP),
indicating that MexAB-independent OprM function likely involves an
efflux process. Moreover, pretreatment of cells with CCCP prior to the accumulation assay abrogated any differences in accumulation levels between the MexAB
OprM+ and
MexAB
OprM
strains, indicating that reduced
drug accumulation by the OprM+ strain (in the absence of
CCCP) cannot be due to OprM-mediated reduction in outer membrane
permeability. It appears, therefore, that OprM can be expressed and
function in a drug efflux capacity independent of MexAB.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Queen's University, Kingston, Ontario K7L 3N6, Canada. Phone: (613) 545-6677. Fax: (613) 545-6796. E-mail: poolek{at}post.queensu.ca.
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