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Papers In Press, published online ahead of print June 24, 2003
J. Biol. Chem, 10.1074/jbc.M306291200
Submitted on June 15, 2003
Revised on June 24, 2003
Accepted on June 24, 2003

Genome-wide expression profiling of the response to polyene, pyrimidine, azole, and echinocandin antifungal agents in Saccharomyces cerevisiae

Agarwal Ameeta K, P. David Rogers, Scott R. Baerson, Melissa R. Jacob, Barker Katherine S, John D. Cleary, Larry A. Walker, Dale G. Nagle, and Alice M. Clark

University of Mississippi, University, Mississippi 38677

Corresponding Author: aagarwal{at}olemiss.edu

Antifungal compounds exert their activity through a variety of mechanisms, some of which are poorly understood. Novel approaches to characterize the mechanism of action of antifungal agents will be of great use in the antifungal drug development process. The aim of the present study was to investigate the changes in the gene expression profile of Saccharomyces cerevisiae following exposure to representatives of the four currently available classes of antifungal agents used in the management of systemic fungal infections. Microarray analysis indicated differential expression of 0.8%, 4.1%, 3.0% and 2.6% of the genes represented on the Affymetrix S98 Yeast gene array in response to ketoconazole, amphotericin B, caspofungin and 5-fluorocytosine (5-FC) respectively. Quantitative real-time RT-PCR was used to confirm the microarray analyses. Genes responsive to ketoconazole, caspofungin, and 5-FC were indicative of the drug-specific effects. Ketoconazole exposure primarily affected genes involved in ergosterol biosynthesis and sterol uptake, caspofungin exposure affected genes involved in cell wall integrity, and 5-FC affected genes involved in DNA and protein synthesis, DNA damage repair, and cell cycle control. In contrast, amphotericin B elicited changes in gene expression reflecting cell stress, membrane reconstruction, transport, phosphate uptake, and cell wall integrity. Genes with the greatest specificity for a particular drug were grouped together as drug-specific genes, whereas genes with a lack of drug specificity were also identified. Taken together, these data shed new light on the mechanisms of action of these classes of antifungal agents and demonstrate the potential utility of gene expression profiling in antifungal drug development.


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