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Journal of Bacteriology, February 2006, p. 1120-1133, Vol. 188, No. 3
0021-9193/06/$08.00+0     doi:10.1128/JB.188.3.1120-1133.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Overexpression of Genes of the Cell Wall Stimulon in Clinical Isolates of Staphylococcus aureus Exhibiting Vancomycin-Intermediate- S. aureus-Type Resistance to Vancomycin

Fionnuala McAleese,1 Shang Wei Wu,2 Krzysztof Sieradzki,2 Paul Dunman,1,{dagger} Ellen Murphy,3 Steven Projan,4 and Alexander Tomasz2*

Wyeth Research, Pearl River, New York,1 Wyeth Research, Cambridge, Massachusetts,4 Wyeth Vaccines, Pearl River, New York,3 Laboratory of Microbiology, The Rockefeller University, New York, New York2

Received 28 September 2005/ Accepted 5 November 2005

Custom-designed gene chips (Affymetrix) were used to determine genetic relatedness and gene expression profiles in Staphylococcus aureus isolates with increasing MICs of vancomycin that were recovered over a period of several weeks from the blood and heart valve of a patient undergoing extensive vancomycin therapy. The isolates were found to be isogenic as determined by the GeneChip based genotyping approach and thus represented a unique opportunity to study changes in gene expression that may contribute to the vancomycin resistance phenotype. No differences in gene expression were detected between the parent strain, JH1, and JH15, isolated from the nares of a patient contact. Few expression changes were observed between blood and heart valve isolates with identical vancomycin MICs. A large number of genes had altered expression in the late stage JH9 isolate (MIC = 8 µg/ml) compared to JH1 (MIC = 1 µg/ml). Most genes with altered expression were involved in housekeeping functions or cell wall biosynthesis and regulation. The sortase-encoding genes, srtA and srtB, as well as several surface protein-encoding genes were downregulated in JH9. Two hypothetical protein-encoding genes, SAS016 and SA2343, were dramatically overexpressed in JH9. Interestingly, 27 of the genes with altered expression in JH9 grown in drug-free medium were found to be also overexpressed when the parental strain JH1 was briefly exposed to inhibitory concentrations of vancomycin, and more than half (17 of 27) of the genes with altered expression belonged to determinants that were proposed to form part of a general cell wall stress stimulon (S. Utaida et al., Microbiology 149:2719-2732, 2003).

* Corresponding author. Mailing address: The Rockefeller University, 1230 York Ave., New York, NY 10021. Phone: (212) 327-8278. Fax: (212) 327-8688. E-mail: tomasz{at}rockefeller.edu.

{dagger} Present address: Department of Microbiology and Pathology, University of Nebraska Medical Center, Omaha, NE 68198.

Journal of Bacteriology, February 2006, p. 1120-1133, Vol. 188, No. 3
0021-9193/06/$08.00+0     doi:10.1128/JB.188.3.1120-1133.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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