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Journal of Bacteriology, July 2005, p. 4372-4380, Vol. 187, No. 13
0021-9193/05/$08.00+0     doi:10.1128/JB.187.13.4372-4380.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Regulation of Pseudomonas Quinolone Signal Synthesis in Pseudomonas aeruginosa

Dana S. Wade, M. Worth Calfee, Edson R. Rocha, Elizabeth A. Ling, Elana Engstrom, James P. Coleman, and Everett C. Pesci*

Department of Microbiology and Immunology, The Brody School of Medicine at East Carolina University, 600 Moye Blvd., Greenville, North Carolina 27834

Received 1 February 2005/ Accepted 29 March 2005

Pseudomonas aeruginosa is an opportunistic pathogen that causes chronic lung infections in cystic fibrosis patients and is a major source of nosocomial infections. This bacterium controls many virulence factors by using two quorum-sensing systems, las and rhl. The las system is composed of the LasR regulator protein and its cell-to-cell signal, N-(3-oxododecanoyl) homoserine lactone, and the rhl system is composed of RhlR and the signal N-butyryl homoserine lactone. A third intercellular signal, the Pseudomonas quinolone signal (PQS; 2-heptyl-3-hydroxy-4-quinolone), also regulates numerous virulence factors. PQS synthesis requires the expression of multiple operons, one of which is pqsABCDE. Previous experiments showed that the transcription of this operon, and therefore PQS production, is negatively regulated by the rhl quorum-sensing system and positively regulated by the las quorum-sensing system and PqsR (also known as MvfR), a LysR-type transcriptional regulator protein. With the use of DNA mobility shift assays and ß-galactosidase reporter fusions, we have studied the regulation of pqsR and its relationship to pqsA, lasR, and rhlR. We show that PqsR binds the promoter of pqsA and that this binding increases dramatically in the presence of PQS, implying that PQS acts as a coinducer for PqsR. We have also mapped the transcriptional start site for pqsR and found that the transcription of pqsR is positively regulated by lasR and negatively regulated by rhlR. These results suggest that a regulatory chain occurs where pqsR is under the control of LasR and RhlR and where PqsR in turn controls pqsABCDE, which is required for the production of PQS.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, East Carolina University School of Medicine, BT 132, 600 Moye Blvd., Greenville, NC 27834. Phone: (252) 744-2351. Fax: (252) 744-3535. E-mail: pescie{at}mail.ecu.edu.

Journal of Bacteriology, July 2005, p. 4372-4380, Vol. 187, No. 13
0021-9193/05/$08.00+0     doi:10.1128/JB.187.13.4372-4380.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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