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Journal of Bacteriology, March 2001, p. 2151-2155, Vol. 183, No. 6
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.6.2151-2155.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Analysis of Promoters Recognized by PvdS, an Extracytoplasmic-Function Sigma Factor Protein from Pseudomonas aeruginosa

Megan J. Wilson, Brendan J. McMorran,dagger and Iain L. Lamont*

Department of Biochemistry, University of Otago, Dunedin, New Zealand

Received 21 August 2000/Accepted 30 November 2000

The alternative sigma factor PvdS is required by Pseudomonas aeruginosa for initiation of transcription from pyoverdine (pvd) promoters. Two divergent PvdS-dependent promoters (pvdE and pvdF) were characterized by deletion analysis, and the minimal promoter region for each included a sequence element, the iron starvation (IS) box, that is present in other pvd promoters. Site-directed mutagenesis showed that the IS box elements were essential for promoter activity in vivo. Band shift assays and in vitro transcription experiments showed that a complex of PvdS and core RNA polymerase required the presence of an IS box in order to bind to and initiate transcription from pvd promoters. These results indicate that IS box elements participate in sequence-specific recognition by PvdS to enable initiation of transcription from pvd promoters and are likely to represent a -35 sequence element for this sigma factor.

* Corresponding author. Mailing address: Department of Biochemistry, University of Otago, P.O. Box 56, Dunedin, New Zealand. Phone: 64 3 479 7869. Fax: 64 3 479 7866. E-mail: iain.lamont{at}stonebow.otago.ac.nz.

dagger Present address: Centre for Molecular Biology and Biotechnology, University of Queensland, Brisbane, Queensland, Australia.

Journal of Bacteriology, March 2001, p. 2151-2155, Vol. 183, No. 6
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.6.2151-2155.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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