GENETIC ANALYSIS OF MULTIDRUG RESISTANCE GENES

多重耐药基因的遗传分析

• 批准号: 3838501
• 负责人: M DEAN
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3838501
• 关键词: Drosophilidae; Escherichia coli; P glycoprotein; Saccharomyces cerevisiae; adenosine triphosphate; binding proteins; chromosomes; genetic mapping; hydropathy; molecular cloning; molecular oncology; multidrug resistance; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; open reading frames; polymerase chain reaction; protein sequence

The development of resistance to chemotherapy drugs is a major limitation in the success of cancer treatment. Particularly problematic is the establishment of multidrug resistance (MDR); due to overexpression of the P-glycoprotein/MDR genes. The MDR genes encode a protein that is a member of a superfamily of membrane-bound transport molecules. This class of proteins contains a pair of adenosine triphosphate (ATP)-binding domains and a pair of hydrophobic domains. The ATP-dependent pump superfamily includes many prokaryotic permeases, the yeast mating factor transporter, the cystic fibrosis gene, and the HLA peptide transporter. MDR genes are capable of pumping a variety of hydrophobic drugs out of cells, including colchicine, vinblastine, and vincristine. We have designed a set of degenerate oligonucleotides to the ATP-binding domain of the MDR genes. These primers have been used in polymerase chain reactions to amplify sequences from Drosophila, Saccharomyces cerevisiae, and Escherichia coli. These products were cloned and sequenced. Each unique clone contained an open reading frame that had homology to the MDR genes, and no clone showed identity to any previously characterized genes. One clone from each species has been used to isolate genomic clones. The Drosophila sequence was mapped to chromosome 2, band 50; adjacent to the Hsc-5 gene. The yeast sequence maps to chromosome 7. The entire sequence has been determined and it contains a hydrophobic domain with 25% amino acid identity to human MDR1 and an ATP-binding domain 45% identical to human MDR1. The E. coli sequence contains a pair of ATP-binding domains 38-45% identical to human MDR1. Genetic experiments are in progress to try to determine the function of these new MDR-related genes. If these genes are capable of transporting drugs, they could prove to be useful models for better understanding the function of MDR genes, for designing effective chemotherapy agents, and for identifying inhibitors of MDR proteins.

对化疗药物产生耐药性是一个主要限制 癌症治疗的成功。 特别有问题的是 建立多药耐药（MDR）;由于过度表达 P-糖蛋白/MDR基因。 MDR基因编码一种蛋白质， 膜结合转运分子超家族的成员 这类 蛋白质含有一对三磷酸腺苷（ATP）结合结构域 和一对疏水结构域。 ATP依赖性泵超家族 包括许多原核透膜酶，酵母交配因子转运蛋白， 囊性纤维化基因和HLA肽转运蛋白。 MDR基因是 能够将各种疏水性药物泵出细胞，包括 秋水仙碱、长春碱和长春新碱。 我们设计了一套 简并寡核苷酸与MDR基因的ATP结合结构域结合。 这些引物已用于聚合酶链反应，以扩增 来自果蝇、酿酒酵母和大肠杆菌的序列。 这些产物被克隆和测序。 每个独特的克隆体都包含一个 开放阅读框与MDR基因具有同源性，没有克隆显示 与任何先前表征的基因相同。 每个克隆一个 物种已被用于分离基因组克隆。 果蝇序列 定位于第2号染色体第50带，与Hsc-5基因相邻。 酵母 序列映射到7号染色体。 整个序列已经确定， 它含有一个与人MDR 1具有25%氨基酸同一性的疏水结构域 和与人MDR 1 45%相同的ATP结合结构域。 急诊杆菌 序列含有一对ATP结合结构域，与人的ATP结合结构域38-45%相同。 MDR1。 基因实验正在进行中，试图确定其功能 这些新的耐多药相关基因。 如果这些基因能够 药物，它们可以被证明是更好地理解 MDR基因的功能，设计有效的化疗药物， 鉴定MDR蛋白的抑制剂。