Hernáez M L, Gil C, Pla J, Nombela C
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Spain.
Yeast. 1998 Apr 30;14(6):517-26. doi: 10.1002/(SICI)1097-0061(19980430)14:6<517::AID-YEA250>3.0.CO;2-D.
The Candida albicans CDR1 gene encodes a member of the ABC-type family of multidrug transporters which has been shown to be involved in azole resistance. Using an in-frame gene fusion between the CDR1 open reading frame and the green fluorescent protein allele yEGFP3, an optimized derivative for its use in C. albicans, we show here how the CDR1-yEGFP3 gene expression is induced in response to azoles as well as to other structurally unrelated drugs like cycloheximide. Moderate increases were observed for calcofluor, canavanine, 5'-fluorcytosine, cilofungin and caffeine, while no induction was found for the antifungals benomyl and amphotericin B or hydrogen peroxide at subinhibitory concentrations. The use of confocal microscopy enabled us to localize the Cdr1p fusion protein at the cell periphery, thus suggesting a cytoplasmic membrane localization. These results suggest deregulation of CDR1 gene as a putative mechanism for the generation of azole resistance in this clinically important pathogenic fungus.
白色念珠菌CDR1基因编码ABC型多药转运蛋白家族的一个成员，该成员已被证明与唑类耐药有关。我们利用CDR1开放阅读框与绿色荧光蛋白等位基因yEGFP3（一种优化的用于白色念珠菌的衍生物）之间的框内基因融合，展示了CDR1 - yEGFP3基因表达如何响应唑类以及其他结构不相关的药物（如环己酰亚胺）而被诱导。对于荧光增白剂、刀豆氨酸、5'-氟胞嘧啶、西洛芬净和咖啡因，观察到适度增加，而在亚抑制浓度下，未发现抗真菌药苯菌灵、两性霉素B或过氧化氢有诱导作用。共聚焦显微镜的使用使我们能够将Cdr1p融合蛋白定位在细胞周边，因此表明其定位于细胞质膜。这些结果表明CDR1基因失调是这种临床上重要的致病真菌产生唑类耐药的一种假定机制。
