Molecular Genetic Analysis of Pdr5p, a Major Yeast Multidrug Transporter

主要酵母多药转运蛋白 Pdr5p 的分子遗传学分析

• 批准号: 8609939
• 负责人: John Golin
• 金额: $ 28.98万
• 依托单位: CATHOLIC UNIVERSITY OF AMERICA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8609939
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; ATP-Binding Cassette Transporters; Amino Acids; Anti-Bacterial Agents; Antibiotic Resistance; Antifungal Agents; Binding; Binding Sites; Biochemical; Biological Assay; Candida; Candidate Disease Gene; Catalytic Domain; Cell membrane; Cells; Chemicals; Clinical; Communication; Complex; Distant; Drug Efflux; Drug Transport; Drug resistance; Funding; Gene Amplification; Genes; Genetic; Hydrolysis; Immune; Infection; Investigation; Lead; Learning; Location; Malignant Neoplasms; Maps; Measures; Mediating; Modeling; Molecular Genetics; Mutation; Nucleotides; Organism; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Play; Poison; Preparation; Proteins; Publications; Pump; Reducing Agents; Regulation; Resistance; Role; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Structure; Suppressor Mutations; Transmembrane Domain; Transport Vesicles; Walkers; Work; Yeasts; antitumor agent; design; deviant; efflux pump; fungus; genetic analysis; improved; interest; killings; member; multi drug transporter; mutant; overexpression; public health relevance; transmission process

PROJECT SUMMARY ABC transporters are found in all known organisms. Overexpression of ABC multidrug transporters is a major cause of clinical resistance to antibiotics, antifungal and antitumor agents. We use the yeast Pdr5 multidrug efflux pump as a model to study an important subfamily of efflux pumps found only in fungi including the highly pathogenic Candida and Cryptocoocus species. In particular we are interested in learning how these complex, polytopic proteins hydrolyze ATP in the nucleotide-binding domains and then use the resulting chemical energy to transport drugs from the transmembrane domains located a good distance away. We used a combination of suppressor genetics, site-directed mutagenesis, and biochemical assays to begin to identify a transmission signal pathway of amino acid residues and will continue that work as described in this proposal to determine whether the pathway we identify by functional studies is similar to the one proposed by purely structural studies of more conventional transporters. If this is the case, such a result would suggest that this interface is conserved among even evolutionary distant drug transporters. Such an observation might lead to the design of agents that reduce pump activity during treatment.

项目摘要 ABC转运蛋白存在于所有已知的生物体中。ABC的过度表达 多药转运蛋白是临床上对抗生素产生耐药性的主要原因， 抗真菌剂和抗肿瘤剂。我们使用酵母Pdr5多药外排泵作为 一个研究仅在真菌中发现的外排泵重要亚家族的模型 包括高致病性念珠菌属和隐球菌属。特别是 我们有兴趣了解这些复杂的多位蛋白质如何水解 ATP的核苷酸结合域，然后使用产生的化学 能量转运药物从跨膜结构域位于一个良好的 距离。我们结合了抑制基因， 诱变和生物化学测定以开始鉴定传输信号 氨基酸残基的途径，并将继续如本文所述的工作， 建议确定我们通过功能研究确定的途径是否 类似于更传统的纯结构研究所提出的一个 运输机如果是这种情况，这样的结果将表明该接口是 甚至在进化遥远的药物转运蛋白中也是保守的。这样的 观察结果可能导致设计出减少泵活性的药物， 治疗