Differential expression of topoisomerase I and RAD52 protein in yeast reveals new facets of the mechanism of action of bisdioxopiperazine compounds.
Differential expression of topoisomerase I and RAD52 protein in yeast reveals new facets of the mechanism of action of bisdioxopiperazine compounds.
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酵母中拓扑异构酶 I 和 RAD52 蛋白的差异表达揭示了双二氧代哌嗪化合物作用机制的新方面。
DOI：
10.1038/sj.bjc.6690767
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发表时间：
1999-11
影响因子：
8.8
通讯作者：
Hill, B T
中科院分区：
文献类型：
--
作者：
van Hille, B;Clerc, X;Creighton, A M;Hill, B T
作为我们新药发现计划的一部分，我们已经建立了一种筛选程序，可以根据化合物直接针对活体酿酒酵母中特定生物靶点的活性来鉴定化合物。该测定的使用提供了第一个直接证据，表明 TOP1 和 RAD52 蛋白参与双二氧代哌嗪 ICRF 化合物的作用模式，因此其表达的作用模式与评估的其他已知 TOP2 抑制剂非常不同。功能测定基于对表型因特定特征而不同的酵母对的比较：异位人类 DNA 拓扑异构酶 I 的表达或缺乏表达，有或没有 RAD52 基因的表达。在一系列抗癌剂中，在表达人拓扑异构酶 I 的酵母中鉴定出拓扑异构酶 I（喜树碱）抑制剂，而 RAD52 蛋白的存在或不存在允许区分产生双链 DNA 断裂的化合物，直接（博来霉素）或涉及 DNA 加合物形成（顺铂），或通过抑制拓扑异构酶 II 酶（依托泊苷）介导的间接 DNA 损伤。然而，值得注意的是，RAD52 蛋白和 TOP1 酶的缺乏似乎与所测试的一系列双二氧代哌嗪 ICRF 化合物的细胞毒性活性有关。因此，这种活体功能测定似乎为探索不同抗癌药物独特且特定的作用模式提供了有价值的工具。 © 1999 癌症研究运动
A screening procedure which permits identification of compounds based on their activities against specific biological targets directly in a living organism, Saccharomyces cerevisiae, has been established as part of our new drug discovery programme. Use of this assay has provided the first direct evidence that TOP1 and RAD52 proteins are involved in the mode of action of bisdioxopiperazine ICRF compounds, which thus express a mode of action quite distinctive from the other known TOP2 inhibitors evaluated. The functional assay is based on a comparison of pairs of yeast differing in their phenotypes by specific traits: the expression or lack of expression of ectopic human DNA topoisomerase I, with or without that of the RAD52 gene. Amongst a series of anticancer agents, inhibitors of topoisomerase I (camptothecin) were identified as such in yeast expressing human topoisomerase I, whilst the presence or absence of RAD52 protein permitted the discrimination of compounds generating double-stranded DNA breaks, either directly (bleomycin) or involving DNA adduct formation (cisplatin), or indirectly with DNA damage mediated via inhibition of the topoisomerase II enzyme (etoposide). Notably, however, both the RAD52 protein and the lack of TOP1 enzyme appeared implicated in the cytotoxic activities of the series of bisdioxopiperazine ICRF compounds tested. This functional assay in a living organism therefore appears to provide a valuable tool for probing distinctive and specific mode(s) of action of diverse anticancer agents. © 1999 Cancer Research Campaign