ANTITOPOISOMERASE DRUG ACTION IN YEAST

描述：DNA拓扑异构酶是广泛的 临床上有用的抗癌药物。虽然基本的生物化学 拓扑异构酶的反应途径是相当清楚的，有 关于该组织的作用机制的信息仍然很少 抗拓扑异构酶药物在生成稳定的共价复合体中的作用 是杀死细胞所必需的。建议开展详细的研究 真核细胞拓扑异构酶II的突变分析 确定酶中与复合稳定剂相互作用的部位 拓扑异构酶II抑制剂。以酵母菌为模型系统进行分析 突变蛋白的作用，以及突变蛋白的过表达 蛋白质。研究策略涉及小说的建构 拓扑异构酶突变体，包括对 抗拓扑异构酶药物。除了依托泊苷，研究将集中在 其他与拓扑异构酶II有不同类型相互作用的药物，如 如ICRF-187，它是该酶的催化抑制物。突变将会 也被引入到人类拓扑异构酶IIα中，并在基因上 验证酵母酶和酵母酶之间突变的影响是相似的 人类的蛋白质。正在开发一种新的系统来调查 拓扑异构酶突变在获得性药物开发中的作用 抵抗。最近的发现表明，拓扑异构酶I可能是一种 其他DNA损伤剂杀死细胞的重要决定因素导致了 研究细胞如何在DNA损伤后调节拓扑异构酶。 这项工作旨在提供对生物化学机制的见解 抗拓扑异构酶药物的作用，并可能在开发新的有用的 和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.