DNA microarray analysis of fluconazole resistance in a laboratory Candida albicans strain  

作　　者：Lan Yan Jundong Zhang Miaohai Li Yongbing Cao Zheng Xu Yingying Cao Pinghui Gao Yan Wang Yuanying Jiang 

机构地区：[1]Department of Pharmacology, College of Pharmacy, Second Military Medical University, Shanghai 200433, China [2]Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang 110016, China

出　　处：《Acta Biochimica et Biophysica Sinica》2008年第12期1048-1060,共13页生物化学与生物物理学报（英文版）

基　　金：This work was supported by the grants from the National Basic Research Program of China （No. 2005CB523105）, the Hi-tech Research and Development Program of China （No. 2007AA02Z187）, the Natural Science Foundation of Shanghai （No. 07ZR14142）, and the Key Programs of Science and Technique Foundation of Shanghai （No. 07JC14064）

摘　　要：Several mechanisms are responsible for the acquired fluconazole （FLC） resistance in Candida albicans. In this study, we developed a FLC-resistant C. albicans strain through serial cultures of a FLC-susceptible C. albicans strain with inhibitory concentrations of FLC. Complimentary DNA microarray analysis and real-time reverse transcription-polymerase chain reaction were used to investigate gene expression changes during the acquisition of azole resistance in the susceptible parental strain and the resistant daughter strain. The differentially expressed genes represented functions as diverse as transporters （e.g. CDR1, PDR17）, ergosterol biosynthesis （e.g. ERG2, ERG9）, sterol metabolism （e.g. ARE2, IPF6464）, energy metabolism （e.g. ADH3, AOX2） and transcription factors （e.g. FCR1, ECM22）. Functional analysis revealed that energy-dependent effiux activity of membrane transporters increased and that ergosterol content decreased with the accumulation of sterol intermediates in the resistant strain as compared with the susceptible strain. We found that a point mutation （N977K） in transcription factor TACI that resulted in hyperactivity of Tacl could be the reason for overexpression of CDR1, CDR2, and PDR17 in the resistant strain. Furthermore, a single amino acid difference （D19E） in ERG3 that led to the inactivation of Erg3 could account for both sterol precursor accumulation and the changes in the expression of ergosterol biosynthesis genes in this resistant strain. These findings expand the understanding of potential novel molecular targets of FLC resistance in clinical C.albicans isolates.Several mechanisms are responsible for the acquired fluconazole （FLC） resistance in Candida albicans. In this study, we developed a FLC-resistant C. albicans strain through serial cultures of a FLC-susceptible C. albicans strain with inhibitory concentrations of FLC. Complimentary DNA microarray analysis and real-time reverse transcription-polymerase chain reaction were used to investigate gene expression changes during the acquisition of azole resistance in the susceptible parental strain and the resistant daughter strain. The differentially expressed genes represented functions as diverse as transporters （e.g. CDR1, PDR17）, ergosterol biosynthesis （e.g. ERG2, ERG9）, sterol metabolism （e.g. ARE2, IPF6464）, energy metabolism （e.g. ADH3, AOX2） and transcription factors （e.g. FCR1, ECM22）. Functional analysis revealed that energy-dependent effiux activity of membrane transporters increased and that ergosterol content decreased with the accumulation of sterol intermediates in the resistant strain as compared with the susceptible strain. We found that a point mutation （N977K） in transcription factor TACI that resulted in hyperactivity of Tacl could be the reason for overexpression of CDR1, CDR2, and PDR17 in the resistant strain. Furthermore, a single amino acid difference （D19E） in ERG3 that led to the inactivation of Erg3 could account for both sterol precursor accumulation and the changes in the expression of ergosterol biosynthesis genes in this resistant strain. These findings expand the understanding of potential novel molecular targets of FLC resistance in clinical C.albicans isolates.

关 键 词：Candida albicans FLUCONAZOLE RESISTANCE MICROARRAY 

分 类 号：Q94[生物学—植物学]