GENETIC ANALYSIS OF PLEIOTROPIC DRUG RESISTANCE

说明(改编自摘要)。 多重耐药指的是获得广泛的 抗性表型是通过少数基因座的遗传变化实现的。 多药耐药是化疗中的一个临床问题 肿瘤和传染病。申请人正在研究多效性。 酿酒酵母耐药(PDR)模型研究 真核多药耐药。以前的工作已经证明 锌指转录因子Pdr1p是导致 细胞对一系列其他有毒化合物的耐受能力。Pdr1p 通过转录激活几个三磷酸腺苷来实现这一功能 编码PDR5和YOR1等基因的结合盒转运体。A基因 最近克隆了一个编码Hsp70同源基因的基因，它调控着 Pdr1p活性。该Hsp70同源物(Pdr13p)可上调Pdr1p 功能，从而增加Pdr1p靶基因的表达 相关的耐药性。这项建议的目标是理解 Pdr13p调控Pdr1p活性的分子细节。 已经制备了针对Pdr13p和Pdr1p的抗血清。我们的计划是 使用此血清在细胞内定位这些因素，以确定是否 这些蛋白质很可能直接相互作用。以及它的影响 在Pdr1p上，Pdr13p还有其他的蛋白质靶点。这些其他靶蛋白 将使用两种混合沙子免疫共沉淀进行鉴定 接近了。Hsp70蛋白的一个重要功能结构域是其 ATPase结构域。有人建议探索Pdr13p依赖的作用 构建和检测ATPas活性对其生物学功能的影响 所预测的缺乏这种酶功能的突变。这个 需要接受正面监管的Pdr1p地区(S) 来自Pdr13p的信号将通过Pdr1p的缺失突变来映射 编码序列。发现Hsp70蛋白的功能是 所需的正常耐药性提供了一个独特的机会 分析真核热休克蛋白70在遗传易感性疾病中的作用 有机体。此外，继续在理解控制方面取得进展 将为酿酒酵母PDR的研究提供重要的基础模式 人肿瘤细胞的多药耐药及其直接模型 白色念珠菌等病原真菌对多药耐药性的研究。

Description (Adapted from the abstract). Multiple drug resistance refers to the acquisition of broad range of resistance phenotypes through genetic changes at a small number of loci. Multidrug resistance is a clinical problem in chemotherapeutic treatment of tumors and infectious disease. The applicant is studying pleiotropic drug resistance (Pdr) in Saccharomyces cerevisiae as a model of eukaryotic multiple drug resistance. Previous work has demonstrated tha the zinc finger transcription factor Pdr1p is a major contributor to the ability of cells to tolerate a range of otherwise toxic compounds. Pdr1p carries this function through transcriptional activation of several ATP binding cassette transporter encoding genes like PDR5 and YOR1. A gene that encodes a Hsp70 homolog was recently cloned that regulates the activity of Pdr1p. This Hsp70 homologue (Pdr13p) can up-regulate Pdr1p function and thereby increase expression of Pdr1p target genes and associated drug resistance. The goal of this proposal is to understand the molecular details behind Pdr13p modulation of Pdr1p activity. Antisera has been prepared against both Pdr13p and Pdr1p. The plan is use this sera to localize these factors within the cell to determine if these proteins are likely to directly interact. Along with its effect on Pdr1p, Pdr13p has other protein targets. These other target proteins will be identified using two hybrid sand co-immunoprecipitation approaches. An important functional domain in a Hsp70 protein is its ATPase domain. It is proposed to explore the role of Pdr13p-dependent ATPas activity in its biological function through construction and assay of mutations that are predicted ot lack this enzymatic function. The regions(s) of Pdr1p that are required to receive the positive regulatory signal from Pdr13p will be mapped by deletion mutagenesis of the Pdr1p coding sequence. The finding that the function of a Hsp70 protein is required for normal drug resistance has provided a unique opportunity to analyze the action of an eukaryotic Hsp70 in a genetically tractable organism. Additionally, continue progress in understanding the control of Pdr in S. cerevisiae will provide and important basic model for multidrug resistance in human tumor cells and a direct model for multidrug tolerance in pathogenic fungi like Candida albicans.