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

doi:10.1128/JB.183.24.7341-7353.2001

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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^, S. Wu,⁴^, 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 biologicalprocesses. Accordingly, a custom-made Affymetrix GeneChip, constituting>86% of the Staphylococcus aureus genome, was used to identifyopen reading frames that are regulated by agr and/or SarA, thetwo best-studied regulators of the organism's virulence response.RNA extracted from wild-type cells and agr, sarA, and agr sarAmutant cells in the early-, mid-, and late-log and stationaryphases of growth was analyzed. Open reading frames with transcriptionpatterns expected of genes either up- or downregulated in an agr-and/or SarA-dependent manner were identified. Oligonucleotidemicroarray and Northern blot analyses confirmed that the transcriptionof several known virulence genes, including hla (alpha-toxin)and spa (protein A), is regulated by each effector and providedinsights about the regulatory cascades involved in both alpha-hemolysinand protein A expression. Several putative virulence factors werealso identified as regulated by agr and/or SarA. In addition,genes that are involved in several biological processes but whichare difficult to reconcile as playing a direct role in the organism'spathogenesis also appeared to be regulated by each effector, suggestingthat products of both the agr and the sarA locus are more-globaltranscription regulators than previouslyrealized.

^* 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.

Present address: Millennium Predictive Medicine, Cambridge, MA02139.

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

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