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Journal of Bacteriology, May 1999, p. 2846-2851, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

The Staphylococcus aureus rsbW (orf159) Gene Encodes an Anti-Sigma Factor of SigB

Eishi Miyazaki,1,2 Jong-Min Chen,1,2 Chiew Ko,1 and William R. Bishai1,2,3,*

Center for Tuberculosis Research, Department of International Health,1 and Department of Molecular Microbiology and Immunology,2 Johns Hopkins School of Public Health, and Division of Infectious Diseases, Department of Medicine, Johns Hopkins School of Medicine,3 Baltimore, Maryland 21205-2179

Received 29 December 1998/Accepted 22 February 1999

SigB, a newly discovered alternative sigma factor of Staphylococcus aureus, has been shown to play an important role in stress responses and the regulation of virulence factors. The rsbW (orf159) gene is immediately upstream of sigB. Its gene product is homologous to Bacillus subtilis RsbW which under appropriate conditions binds to B. subtilis SigB and functions as an anti-sigma factor or negative posttranslational regulator. To define the function of S. aureus RsbW, both the S. aureus SigB and RsbW proteins were expressed in Escherichia coli and purified. Cross-linking experiments with these purified proteins revealed that RsbW was capable of specific binding to SigB. In an in vitro transcription runoff assay, RsbW prevented SigB-directed transcription from the sar P3 promoter, a known SigB-dependent promoter, and the inhibitory activity of RsbW was found to be concentration dependent. We also identified SigB promoter consensus sequences upstream of the genes encoding alkaline shock protein 23 and coagulase and have demonstrated SigB and RsbW dependence for the promoters in vitro. These results show that RsbW is a protein sequestering anti-sigma factor of S. aureus SigB and suggest that SigB activity in S. aureus is regulated posttranslationally.

* Corresponding author. Mailing address: Center for Tuberculosis Research, Johns Hopkins School of Public Health, 615 N. Wolfe St., Baltimore, MD 21205-2179. Phone: (410) 955-3507. Fax: (410) 614-8173. E-mail: wbishai{at}jhsph.edu.

Journal of Bacteriology, May 1999, p. 2846-2851, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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