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Journal of Bacteriology, August 2004, p. 5267-5280, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5267-5280.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification of sarV (SA2062), a New Transcriptional Regulator, Is Repressed by SarA and MgrA (SA0641) and Involved in the Regulation of Autolysis in Staphylococcus aureus

Adhar C. Manna,1* Susham S. Ingavale,1 MaryBeth Maloney,1 Willem van Wamel,2 and Ambrose L. Cheung1

Department of Microbiology, Dartmouth Medical School, Hanover, New Hampshire 03755,1 Department of Medical Microbiology, University of Utrecht, Utrecht, The Netherlands2

Received 24 March 2004/ Accepted 18 May 2004

The expression of genes involved in the pathogenesis of Staphylococcus aureus is known to be controlled by global regulatory loci, including agr, sarA, sae, arlRS, lytSR, and sarA-like genes. Here we described a novel transcriptional regulator called sarV of the SarA protein family. The transcription of sarV is low or undetectable under in vitro conditions but is significantly augmented in sarA and mgrA (norR or rat) (SA0641) mutants. The sarA and mgrA genes act as repressors of sarV expression, as confirmed by transcriptional fusion and Northern analysis data. Purified SarA and MgrA proteins bound specifically to separate regions of the sarV promoter as determined by gel shift and DNase I footprinting assays. The expression of 19 potential target genes involved in autolysis and virulence, phenotypes affected by sarA and mgrA, was evaluated in an isogenic sarV mutant pair. Our data indicated that the sarV gene product played a role regulating some virulence genes and more genes involved in autolysis. The sarV mutant was more resistant to Triton X-100 and penicillin-induced lysis compared to the wild type and the sarA mutant, whereas hyperexpression of sarV in the parental strain or the sarV mutant rendered the resultant strain highly susceptible to lysis. Zymographic analysis of murein hydrolase activity revealed that inactivation of the sarV gene results in decreased extracellular murein hydrolase activity compared to that of wild-type S. aureus. We propose that sarV may be part of the common pathway by which mgrA and sarA gene products control autolysis in S. aureus.

* Corresponding author. Mailing address: Room No. 205, Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, NH 03755. Phone: (603) 650-1310. Fax: (603) 650-1362. E-mail: Adhar.C.Manna{at}Dartmouth.EDU.

Journal of Bacteriology, August 2004, p. 5267-5280, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5267-5280.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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