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Infection and Immunity, February 2001, p. 885-896, Vol. 69, No. 2
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.2.885-896.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Characterization of sarR, a Modulator of sar Expression in Staphylococcus aureus

Adhar Manna and Ambrose L. Cheung*

Department of Microbiology, Dartmouth Medical School, Hanover, New Hampshire 03755

Received 18 August 2000/Returned for modification 18 September 2000/Accepted 17 October 2000

The expression of virulence determinants in Staphylococcus aureus is controlled by global regulatory loci (e.g., sar and agr). The sar locus is composed of three overlapping transcripts (sar P1, P3, and P2 transcripts from P1, P3, and P2 promoters, respectively), all encoding the 372-bp sarA gene. The level of SarA, the major regulatory protein, is partially controlled by the differential activation of sar promoters. We previously partially purified a ~12 kDa protein with a DNA-specific column containing a sar P2 promoter fragment. In this study, the putative gene, designated sarR, was identified and found to encode a 13.6-kDa protein with homology to SarA. Transcriptional and immunoblot studies revealed the sarR gene to be expressed in other staphylococcal strains. Recombinant SarR protein bound sar P1, P2, and P3 promoter fragments in gel shift and footprinting assays. A sarR mutant expressed a higher level of P1 transcript than the parent, as confirmed by promoter green fluorescent protein fusion assays. As the P1 transcript is the predominant sar transcript, we confirmed that the sarR mutant expressed more SarA than the parental strain. We thus proposed that SarR is a regulatory protein that binds to the sar promoters to down-regulate P1 transcription and the ensuing SarA protein expression.

* Corresponding author. Mailing address: Department of Microbiology Vail 206, Dartmouth Medical School Hanover, NH 03755. Phone: (603) 650-1340. Fax: (603) 650-1362. E-mail: ambrose.cheung{at}dartmouth.edu.

Infection and Immunity, February 2001, p. 885-896, Vol. 69, No. 2
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.2.885-896.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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