Identification of genes differentially expressed in association with reduced azole susceptibility in Saccharomyces cerevisiae
Identification of genes differentially expressed in association with reduced azole susceptibility in Saccharomyces cerevisiae
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DOI：
10.1093/jac/dkg217
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发表时间：
2003-05-01
影响因子：
5.2
通讯作者：
Rogers, PD
中科院分区：
文献类型：
--
作者：
Barker, KS;Pearson, MM;Rogers, PD
目的：建立一株对氟康唑和伊曲康唑敏感性降低的酿酒酵母菌株，用于筛选与该表型相关的差异表达基因。伊曲康唑和氟康唑对初始分离株(9763S)的MIC值分别为2和16 mg/L，对最终分离株(97631)的MIC值分别为16和64 mg/L。从9763S和97631中分离出重复的总RNA，并与Affymetrix S98酵母阵列杂交。结果：在芯片上显示的近6400个开放阅读框中，共发现116个差异表达基因(1.8%)。细胞壁维持基因TIR4和CCW12，固醇代谢基因Upc2，小分子运输基因AUS1和YHK8，以及胁迫反应基因CUP1-1的表达水平至少是9763S的2.5倍。11个能量产生基因，离子稳态基因FRE1，FRE2和FRE4，以及甾醇代谢基因ERG8和ERG13的表达水平至少比9763S的表达水平低2.5倍。结论：本研究中发现的几个差异表达基因在真菌对唑处理的反应中存在差异表达。此外，AUS1和/或YHK8作为药物外流的介体的潜在作用是耐人寻味的，值得进一步研究。
Objective: An isolate of S. cerevisiae with reduced susceptibility to fluconazole and itraconazole was developed in the laboratory and used to identify genes that are differentially expressed in association with this phenotype.Methods: S. cerevisiae strain ATCC 9763 was passaged in increasing concentrations of itraconazole. Itraconazole and fluconazole MiCs for the initial isolate (9763S) were 2 and 16 mg/L and for the final isolate (97631) were 16 and greater than or equal to64 mg/L, respectively. Duplicate sets of total RNA from 9763S and 97631 were isolated and hybridized to Affymetrix S98 yeast arrays. To validate results, six differentially expressed genes were further examined by RT-PCR.Results: Of the nearly 6400 open reading frames represented on the array, a total of 116 genes (1.8%) were found to be differentially expressed. Cell wall maintenance genes TIR4 and CCW12, sterol metabolism gene UPC2, small molecule transport genes AUS1 and YHK8, and stress response gene CUP1-1 were expressed at a level at least 2.5-fold higher than the expression level found in 9763S. Eleven energy generation genes, ionic homeostasis genes FRE1, FRE2and FRE4, and sterol metabolism genes ERG8 and ERG13 were expressed at least 2.5-fold lower than the expression level found in 9763S.Conclusions: Several genes found to be differentially expressed in this study have been shown previously to be differentially expressed in the fungal response to azole treatment. In addition, the potential role of AUS1 and/or YHK8 as mediators of drug efflux is intriguing and warrants further study.