ANTITOPOISOMERASE DRUG ACTION IN YEAST

描述：DNA 拓扑异构酶是多种药物的靶标 临床上有用的抗癌药物。 虽然有基础的生化 拓扑异构酶的反应途径已相当清楚，有 关于其作用机制的信息仍然很少 抗拓扑异构酶药物产生稳定的共价复合物，即 杀死细胞所必需的。 建议开展详细研究 真核拓扑异构酶 II 的突变分析，目的是 识别酶中与复合物稳定相互作用的位点 拓扑异构酶 II 抑制剂。 使用酵母作为模型系统进行分析 突变蛋白的影响以及突变体的过度表达 蛋白质。 研究策略涉及新颖的构建 拓扑异构酶突变体，包括对拓扑异构酶过敏的突变体 抗拓扑异构酶药物。 除依托泊苷外，研究还将集中于 与拓扑异构酶 II 具有不同类型相互作用的其他药物，例如 如 ICRF-187，它是该酶的催化抑制剂。 突变会 也可被引入人类拓扑异构酶IIα并处于基因中 验证酵母酶中突变的影响是相似的，并且 人类蛋白质。 正在开发一个新系统来调查是否 拓扑异构酶突变在获得性药物的开发中发挥作用 阻力。 最近的研究结果表明拓扑异构酶 I 可能是 其他 DNA 损伤剂杀死细胞的重要决定因素 研究细胞如何在 DNA 损伤后调节拓扑异构酶。 这项工作旨在深入了解 抗拓扑异构酶药物作用，可能有助于开发新的药物 以及更有效的拓扑异构酶抑制剂。

DESCRIPTION: DNA topoisomerases are the targets for a wide range of clinically useful anti-cancer drugs. Although the basic biochemical reaction pathway for the topoisomerases is fairly well understood, there is still little information available regarding the mechanism of action of the anti-topoisomerase drugs in generating the stable covalent complex that is necessary for cell killing. Studies are proposed to carry out a detailed mutational analysis of eukaryotic topoisomerase II, with the goal of identifying sites in the enzyme which interact with complex stabilizing topoisomerase II inhibitors. Yeast is used as a model system for analyzing the effects of mutant proteins, and for overexpression of the mutant proteins. The research strategy involves the construction of novel topoisomerase mutants, including mutants that are hypersensitive to anti-topoisomerase drugs. In addition to etoposide, studies will focus on other agents with different types of interaction with topoisomerase II, such as ICRF-187, which is a catalytic inhibitor of the enzyme. Mutations will also be introduced into the human topoisomerase II alpha and be at genes to verify that the effects of mutations are similar among the yeast enzymes and the human proteins. A new system is being developed to investigate whether mutations in topoisomerase play a role in the development of acquired drug resistance. Recent findings indicating that topoisomerase I may be an important determinant in cell killing by other DNA damaging agents have led to studies of how cells may modulate topoisomerases following DNA damage. This work is designed to provide insight into the biochemical mechanisms of anti-topoisomerase drug action, and may be useful in the development of new and more effective topoisomerase inhibitors.