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Journal of Bacteriology, March 2001, p. 1843-1852, Vol. 183, No. 6
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.6.1843-1852.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

sigma B Activity Depends on RsbU in Staphylococcus aureus

P. Giachino,1 S. Engelmann,2 and M. Bischoff1,*

Institute of Medical Microbiology, University of Zürich, CH-8028 Zürich, Switzerland,1 and Institute of Microbiology and Molecular Biology, Ernst-Moritz-Arndt University, D-17487 Greifswald, Germany2

Received 6 September 2000/Accepted 14 December 2000

Derivatives of the widely used laboratory strain Staphylococcus aureus NCTC8325, which are natural rsbU mutants, were shown to be unable to produce RsbU, a positive regulator of the alternative sigma factor sigma B. The lack of RsbU prevented the heat-dependent production of sigma B-controlled transcripts and resulted in reduced H2O2 and UV tolerance, enhanced alpha-hemolysin activity, and the inability to produce the alkaline shock protein Asp23. After 48 h of growth, rsbU mutant strains failed to accumulate staphyloxanthin, the major stationary-phase carotenoid. Transcription of Asp23 was found to be exclusively controlled by sigma B, making it an excellent target for the study of sigma B activity in S. aureus. Reporter gene experiments, using the firefly luciferase gene (luc+) fused to the sigma B-dependent promoter(s) of asp23, revealed that sigma B is almost inactive in 8325 derivatives. cis complementation of the 8325 derivative BB255 with the wild-type rsbU gene from strain COL produced the rsbU+ derivative GP268, a strain possessing a sigma B activity profile comparable to that of the rsbU+ wild-type strain Newman. In GP268, the heat inducibility of sigma B-dependent genes, Asp23 production, alpha-hemolysin activity, pigmentation, and susceptibility to H2O2 were restored to the levels observed in strain Newman, clearly demonstrating that RsbU is needed for activation of sigma B in S. aureus.

* Corresponding author. Mailing address: Institute of Medical Microbiology, University of Zürich, Gloriastr. 32, Postfach, CH-8028 Zürich, Switzerland. Phone: 41 1 634 26 70. Fax: 41 1 634 49 06. E-mail: bischoff{at}immv.unizh.ch.

Journal of Bacteriology, March 2001, p. 1843-1852, Vol. 183, No. 6
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.6.1843-1852.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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