JAC Advance Access originally published online on June 16, 2004
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Journal of Antimicrobial Chemotherapy 2004 54(1):46-55; doi:10.1093/jac/dkh245
JAC vol.54 no.1 © The British Society for Antimicrobial Chemotherapy 2004; all rights reserved.
Response of Saccharomyces cerevisiae to a monoterpene: evaluation of antifungal potential by DNA microarray analysis
1 United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3–5–8 Saiwai, Fuchu, Tokyo 183; 2 Research Institute of Biological Resources, 3 Gene Function Research Center, 5 International Patent Organism Depository, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, Higashi 1–1-1, Tsukuba, Ibaraki 305–8566; 4 Daikin Environmental Laboratory, Ltd. 3 Banchi, Miyukigaoka, Tsukuba-shi, Ibaraki, 305–0841 Japan
* Corresponding author. Tel: +81-298-61-6059; Fax: +81-298-61-6066; E-mail: hitoshi.iwahashi{at}aist.go.jp
Plant-derived essential oils with monoterpenoids have been used as antifungal drugs since ancient times, but the mode of action of these natural hydrocarbons at the molecular level is not understood. In order to understand the mechanisms of toxicity of 
-terpinene (a cyclic monoterpene), a culture of Saccharomyces cerevisiae was exposed to 0.02% -terpinene for 2 h and transcript profiles were obtained using yeast DNA arrays. These profiles, when compared with transcript profiles of untreated cultures, revealed that the expression of 793 genes was affected. For 435 genes, mRNA levels in treated cells compared with control cells differed by more than two-fold, whereas for 358 genes, it was <0.5-fold. Northern blots were performed for selected genes to verify the microarray results. Functional analysis of the up-regulated genes indicates that, similar to commonly used antifungal drugs, -terpinene exposure affected genes involved in ergosterol biosynthesis and sterol uptake. In addition, transcriptional induction of genes related to lipid metabolism, cell wall structure and function, detoxification and cellular transport was observed in response to terpinene toxicity. Notably, the functions of 192 up-regulated genes are still unknown, but their characterization will probably shed light on the mechanisms of drug resistance and sensitivity. Taken together, this study showed that -terpinene has strong antifungal activities and its modes of action resemble those of presently used antifungal drugs.
Keywords: essential oils , ergosterol , stress response , toxicity , genomic expression
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