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Journal of Bacteriology, March 2007, p. 2376-2391, Vol. 189, No. 6
0021-9193/07/$08.00+0     doi:10.1128/JB.01439-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Extensive and Genome-Wide Changes in the Transcription Profile of Staphylococcus aureus Induced by Modulating the Transcription of the Cell Wall Synthesis Gene murF{triangledown} ,{dagger}

Rita G. Sobral,1,5 Alison E. Jones,2 Shelley G. Des Etages,2 Thomas J. Dougherty,3 Robert M. Peitzsch,2 Terry Gaasterland,4 Ana Madalena Ludovice,1 Hermínia de Lencastre,1,5 and Alexander Tomasz5*

Molecular Genetics Laboratory, Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa, 2780 Oeiras, Portugal,1 Genetic Technologies, Pfizer Global Research and Development, Groton, Connecticut 06340,2 Antibacterial Biology, Pfizer Global Research and Development, Groton, Connecticut 06342,3 Scripps Institute of Oceanography, La Jolla, California 92093,4 The Rockefeller University, 1230 York Avenue, New York, New York 100215

Received 11 September 2006/ Accepted 17 December 2006

A murF conditional mutant was used to evaluate the effect of suboptimal transcription of this gene on the transcriptome of the methicillin-resistant Staphylococcus aureus strain COL. The mutant was grown in the presence of optimal and suboptimal concentrations of the inducer, and the relative levels of transcription of genes were evaluated genome wide with an Affymetrix DNA microarray that included all open reading frames (ORFs) as well as intergenic sequences derived from four sequenced S. aureus strains. Using a sensitivity threshold value of 1.5, suboptimal expression of murF altered the transcription of a surprisingly large number of genes, i.e., 668 out of the 2,740 ORFs (close to one-fourth of all ORFs), of the genome of S. aureus strain COL. The genes with altered transcription were distributed evenly around the S. aureus chromosome, and groups of genes involved with distinct metabolic functions responded in unique and operon-specific manners to modulation in murF transcription. For instance, all genes belonging to the isd operon and all but 2 of the 35 genes of prophage L54a were down-regulated, whereas all but one of the 21 members of the vraSR regulon and most of the 79 virulence-related genes (those for fibronectin binding proteins A and B, clumping factor A, gamma hemolysin, enterotoxin B, etc.) were up-regulated in cells with suboptimal expression of murF. Most importantly, the majority of these altered gene expression profiles were reversible by resupplying the optimal concentration of IPTG (isopropyl-ß-D-thiogalactopyranoside) to the culture. The observations suggest the coordinate regulation of a large sector of the S. aureus transcriptome in response to a disturbance in cell wall synthesis.

* Corresponding author. Mailing address: The Rockefeller University, 1230 York Avenue, New York, NY 10021. Phone: (212) 327-8277. Fax: (212) 327-8688. E-mail: tomasz{at}mail.rockefeller.edu.

{triangledown} Published ahead of print on 28 December 2006.

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

Journal of Bacteriology, March 2007, p. 2376-2391, Vol. 189, No. 6
0021-9193/07/$08.00+0     doi:10.1128/JB.01439-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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