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Journal of Bacteriology, December 2001, p. 7341-7353, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7341-7353.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Transcription Profiling-Based Identification of Staphylococcus aureus Genes Regulated by the agr and/or sarA Loci

P. M. Dunman,1 E. Murphy,2 S. Haney,1 D. Palacios,1 G. Tucker-Kellogg,3,dagger S. Wu,4,Dagger E. L. Brown,3 R. J. Zagursky,5 D. Shlaes,1 and S. J. Projan1,*

Infectious Diseases1 and Department of Bioinformatics,2 Wyeth-Ayerst Research, Pearl River, New York 10965; Department of Genomics, Wyeth-Ayerst Research, Cambridge, Massachusetts 021403; Wyeth-Ayerst Research, Monmouth Junction, New Jersey 08852-95144; and Wyeth-Lederle Vaccines, West Henrietta, New York 145865

Received 2 July 2001/Accepted 26 September 2001

The advent of transcription profiling technologies has provided researchers with an unprecedented ability to study biological processes. Accordingly, a custom-made Affymetrix GeneChip, constituting >86% of the Staphylococcus aureus genome, was used to identify open reading frames that are regulated by agr and/or SarA, the two best-studied regulators of the organism's virulence response. RNA extracted from wild-type cells and agr, sarA, and agr sarA mutant cells in the early-, mid-, and late-log and stationary phases of growth was analyzed. Open reading frames with transcription patterns expected of genes either up- or downregulated in an agr- and/or SarA-dependent manner were identified. Oligonucleotide microarray and Northern blot analyses confirmed that the transcription of several known virulence genes, including hla (alpha-toxin) and spa (protein A), is regulated by each effector and provided insights about the regulatory cascades involved in both alpha-hemolysin and protein A expression. Several putative virulence factors were also identified as regulated by agr and/or SarA. In addition, genes that are involved in several biological processes but which are difficult to reconcile as playing a direct role in the organism's pathogenesis also appeared to be regulated by each effector, suggesting that products of both the agr and the sarA locus are more-global transcription regulators than previously realized.

* Corresponding author. Mailing address: Wyeth-Ayerst Research, Department of Infectious Diseases, 401 N. Middletown Rd., Bldg. 205, Rm. 286, Pearl River, NY 10965. Phone: (845) 602-3063. Fax: (845) 602-2480. E-mail: projans{at}war.wyeth.com.

dagger Present address: Millennium Predictive Medicine, Cambridge, MA 02139.

Dagger Present address: Bristol-Myers Squibb PRI, Bioinformatics, Princeton, NJ 08543-5400.

Journal of Bacteriology, December 2001, p. 7341-7353, Vol. 183, No. 24
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.24.7341-7353.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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