The yeast ATP binding cassette (ABC) protein genes PDR10 and PDR15 are novel targets for the Pdr1 and Pdr3 transcriptional regulators
The yeast ATP binding cassette (ABC) protein genes PDR10 and PDR15 are novel targets for the Pdr1 and Pdr3 transcriptional regulators
复制标题
DOI：
10.1016/s0014-5793(97)01382-3
复制
发表时间：
1997-12
期刊：
影响因子：
3.5
通讯作者：
H. Wolfger;Y. Mahé;A. Parle‐McDermott;A. Delahodde;K. Kuchler
中科院分区：
文献类型：
--
作者：
H. Wolfger;Y. Mahé;A. Parle‐McDermott;A. Delahodde;K. Kuchler
酵母转录因子Pdr1和Pdr3控制多效性耐药（PDR）的发展，因为它们通过被称为PDREs （R响应元件）的结合作用位点调节ATP结合盒（ABC）药物外排泵的表达。在本报告中，我们通过Northern blotting、凝胶位移迁移率测定和dna酶I足迹分析表明，ABC基因espdr10和pdr15的转录也受到Pdr1和Pdr3的控制。此外，体外带移实验表明，GST - Pdr1融合蛋白可以结合pdr10和pdr15的PDREs。dna酶I足迹允许鉴定精确的PDRE结合基序，表明在pdr15启动子中存在一种新的略微退化的PDRE基序。最后，pdr10和pdr15mrna水平在携带Δpdr1， Δpdr3and Δpdr1Δpdr3deletions orpdr1‐3和pdr3‐2功能增益突变的等基因酵母菌株中丰度变化显著，表明pdr10和pdr15都是酵母PDR网络的新成员。
The yeast transcription factors Pdr1 and Pdr3 control pleiotropic drug resistance (PDR) development, since they regulate expression of ATP‐binding cassette (ABC) drug efflux pumps through binding tocis‐acting sites known as PDREs ( R esponsive lements). In this report, we show by Northern blotting, gel shift mobility assays and DNase I footprinting that transcription of the ABC genesPDR10andPDR15is also controlled by Pdr1 and Pdr3. In addition, in vitro band shift assays demonstrate that a GST‐Pdr1 fusion protein can bind to the PDREs ofPDR10andPDR15. DNase I footprinting allowed the identification of the precise PDRE binding motifs, indicating the presence of a novel slightly degenerate PDRE motif in thePDR15promoter. Finally,PDR10andPDR15mRNA levels vary dramatically in abundance in isogenic yeast strains carrying either Δpdr1, Δpdr3and Δpdr1Δpdr3deletions orpdr1‐3andpdr3‐2gain‐of‐function mutations, demonstrating that bothPDR10andPDR15are new members of the yeast PDR network.