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Journal of Bacteriology, February 2005, p. 1384-1391, Vol. 187, No. 4
0021-9193/05/$08.00+0     doi:10.1128/JB.187.4.1384-1391.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Overexpression of the Multidrug Efflux Pumps MexCD-OprJ and MexEF-OprN Is Associated with a Reduction of Type III Secretion in Pseudomonas aeruginosa

Juan F. Linares, Juan A. López,{dagger} Emilio Camafeita,{dagger} Juan P. Albar, Fernando Rojo, and Jose L. Martínez*

Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco, Madrid, Spain

Received 7 May 2004/ Accepted 9 November 2004

The Pseudomonas aeruginosa genome contains several different multidrug resistance (MDR) efflux pumps. Overproduction of these pumps reduces susceptibility to a variety of antibiotics. Some recently published works have analyzed the effect of the overproduction of MDR efflux pumps on bacterial virulence. Here we have studied the effect of overproduction of the efflux pumps MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY on type III secretion (T3S) in P. aeruginosa. The type III secretion system (T3SS) is used by P. aeruginosa to deliver toxins directly into the cytoplasm of the host cell. Our data indicate that overexpression of either MexCD-OprJ or MexEF-OprN is associated with the impairment of T3S in P. aeruginosa. No effect on overexpression of either MexAB-OprM or MexXY was detected. The observed defect in T3S was due to a lack of expression of genes belonging to the T3SS regulon. Transcription of this regulon is activated by ExsA in response to environmental signals. Overexpression of this transcriptional regulator complemented the defect in T3S observed in the MexCD-OprJ- and MexEF-OprN-overproducing strains. Taken together, these results suggest that overproduction of either MexCD-OprJ or MexEF-OprN is associated with a reduction in the transcription of the T3SS regulon due to the lack of expression of the exsA gene, encoding the master regulator of the system. The relevance of potential metabolic and quorum-sensing imbalances due to overexpression of MDR pumps associated with this phenotype is also discussed.

* Corresponding author. Mailing address: Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone: 34-91-5854542. Fax: 34-91-5854506. E-mail: jlmtnez{at}cnb.uam.es.

{dagger} Present address: Unidad de Proteómica, Fundación Centro Nacional de Investigaciones Cardiovasculares Carlos III, Tres Cantos 28760 Madrid, Spain.

Journal of Bacteriology, February 2005, p. 1384-1391, Vol. 187, No. 4
0021-9193/05/$08.00+0     doi:10.1128/JB.187.4.1384-1391.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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