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Journal of Bacteriology, April 2006, p. 2593-2603, Vol. 188, No. 7
0021-9193/06/$08.00+0     doi:10.1128/JB.188.7.2593-2603.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Characterizing the Effect of the Staphylococcus aureus Virulence Factor Regulator, SarA, on Log-Phase mRNA Half-Lives{dagger}

Corbette Roberts,1 Kelsi L. Anderson,1 Ellen Murphy,2 Steven J. Projan,3 William Mounts,3 Barry Hurlburt,4 Mark Smeltzer,5 Ross Overbeek,6 Terrence Disz,7 and Paul M. Dunman1*

Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198,1 Bacterial Vaccines, Wyeth Research, Pearl River, New York 10965,2 Wyeth Biological Technologies, Cambridge, Massachusetts 02140,3 United States Department of Agriculture, Ag Research Service-Southern Regional Research Center, New Orleans, Louisiana 70124,4 Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205,5 Fellowship for Interpretation of Genomes, Burr Ridge, Illinois 60527,6 Mathematics and Computer Science, Argonne National Laboratory, Argonne, Illinois 604397

Received 7 November 2005/ Accepted 17 January 2006

Bacterial pathogens regulate virulence factor expression at both the level of transcription initiation and mRNA processing/turnover. Within Staphylococcus aureus, virulence factor transcript synthesis is regulated by a number of two-component regulatory systems, the DNA binding protein SarA, and the SarA family of homologues. However, little is known about the factors that modulate mRNA stability or influence transcript degradation within the organism. As our entree to characterizing these processes, S. aureus GeneChips were used to simultaneously determine the mRNA half-lives of all transcripts produced during log-phase growth. It was found that the majority of log-phase transcripts (90%) have a short half-life (<5 min), whereas others are more stable, suggesting that cis- and/or trans-acting factors influence S. aureus mRNA stability. In support of this, it was found that two virulence factor transcripts, cna and spa, were stabilized in a sarA-dependent manner. These results were validated by complementation and real-time PCR and suggest that SarA may regulate target gene expression in a previously unrecognized manner by posttranscriptionally modulating mRNA turnover. Additionally, it was found that S. aureus produces a set of stable RNA molecules with no predicted open reading frame. Based on the importance of the S. aureus agr RNA molecule, RNAIII, and small stable RNA molecules within other pathogens, it is possible that these RNA molecules influence biological processes within the organism.

* Corresponding author. Mailing address: 986495 Nebraska Medical Center, Omaha, NE 68198-6495. Phone: (402) 559-7745. Fax: (402) 559-4077. E-mail: pdunman{at}unmc.edu.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, April 2006, p. 2593-2603, Vol. 188, No. 7
0021-9193/06/$08.00+0     doi:10.1128/JB.188.7.2593-2603.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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