TRANSCRIPTIONAL CONTROL OF THE YEAST PDR5 GENE BY THE PDR3 GENE-PRODUCT
TRANSCRIPTIONAL CONTROL OF THE YEAST PDR5 GENE BY THE PDR3 GENE-PRODUCT
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DOI：
10.1128/mcb.14.7.4653
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发表时间：
1994-07-01
影响因子：
5.3
通讯作者：
MOYEROWLEY, WS
中科院分区：
文献类型：
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作者：
KATZMANN, DJ;BURNETT, PE;MOYEROWLEY, WS
酿酒酵母细胞具有同时对一系列具有不同细胞毒活性的药物产生耐药性的能力。参与这一获取的基因被称为多效性耐药(PDR)基因。已经发现了几个半显性的、编码耐药的PDR突变，它们位于第二染色体着丝粒附近，包括PDR3-1和PDR4-1。染色体II的DNA测序发现了一个潜在的开放阅读框架，命名为YBL03-23，它可能编码一种与PDR1基因产物序列高度相似的蛋白质，PDR1基因是一种含锌指转录因子。在这里，我们证明了YBL03-23与PDR3等位基因。PDR1或PDR3的功能拷贝的存在对于耐药性和可能的膜转运蛋白编码基因PDR5的表达是必不可少的。PDR5启动子的缺失图谱确定了该基因表达所必需的-360到-112区域。利用细菌表达的Pdr3p进行的DNase I足迹分析表明，该蛋白至少能识别PDR5启动子-360/-112区间中的一个位点。携带PDR3基因的高拷贝数质粒提高了对寡霉素和放线菌亚胺的抗性。在Delta pdr5菌株中增加PDR3基因拷贝数增加了对寡霉素的耐药性，但不能纠正由于失去PDR5而导致的放线菌亚胺过敏症。这些数据与以下观点一致，即PDR3通过提高PDR5的转录水平来增加放线菌亚胺的抗性，而PDR3介导的寡霉素抗性则通过其他一些靶基因发挥作用。
Saccharomyces cerevisiae cells possess the ability to simultaneously acquire resistance to an array of drugs with different cytotoxic activities. The genes involved in this acquisition are referred to as pleiotropic drug resistant (PDR) genes. Several semidominant, drug resistance-encoding PDR mutations have been found that map near the centromere on chromosome II, including PDR3-1 and PDR4-1. DNA sequencing of chromosome II identified a potential open reading frame, designated YBL03-23, that has the potential to encode a protein with strong sequence similarity to the product of the PDR1 gene, a zinc finger-containing transcription factor. Here,ve show that YBL03-23 is allelic with PDR3. The presence of a functional copy of either PDR1 or PDR3 is essential for drug resistance and expression of a putative membrane transporter-encoding gene, PDR5. Deletion mapping of the PDR5 promoter identified a region from -360 to -112 that is essential for expression of this gene. DNase I footprinting analysis using bacterially expressed Pdr3p showed specific recognition by this protein of at least one site in the -360/-112 interval in the PDR5 promoter. A high-copy-number plasmid carrying the PDR3 gene elevated resistance to both oligomycin and cycloheximide. Increasing the number of PDR3 gene copies in a Delta pdr5 strain increased oligomycin resistance but was not able to correct the cycloheximide hypersensitivity that results from loss of PDR5. These data are consistent with the notion that PDR3 acts to increase cycloheximide resistance by elevating the level of PDR5 transcription, while PDR3-mediated oligomycin resistance acts through some other target gene.