Chemical Genomics Research Group/Chemical Genetics Laboratory, RIKEN Advanced Science Institute, Wako, Saitama 351-0198, Japan.
Biochem Biophys Res Commun. 2010 Apr 9;394(3):569-73. doi: 10.1016/j.bbrc.2010.03.021. Epub 2010 Mar 7.
The telomere-associated protein tankyrase 1 is a poly(ADP-ribose) polymerase and is considered to be a promising target for cancer therapy, especially for BRCA-associated cancers. However, an efficient assay system for inhibitor screening has not been established, mainly due to the difficulty of efficient preparation of the enzyme and its substrate. Here, we report a cell-based assay system for detecting inhibitory activity against tankyrase 1. We found that overexpression of the human tankyrase 1 gene causes a growth defect in the fission yeast Schizosaccharomyces pombe. Chemicals that restore the growth defect phenotype can be identified as potential tankyrase 1 inhibitors. We performed a high-throughput screen using this system, and identified flavone as a compound that restores the growth of yeast cells overexpressing tankyrase 1. Indeed, flavone inhibited poly(ADP-ribosyl)ation of proteins caused by overexpression of tankyrase 1 in yeast cells. This system allows rapid identification of inhibitory activity against tankyrase 1 and is amenable to high-throughput screening using robotics.
端粒相关蛋白 tankyrase 1 是一种多聚（ADP-核糖）聚合酶，被认为是癌症治疗的一个有前途的靶点，特别是对 BRCA 相关癌症。然而，由于酶及其底物的高效制备存在困难，因此尚未建立有效的抑制剂筛选测定系统。在这里，我们报告了一种用于检测 tankyrase 1 抑制活性的基于细胞的测定系统。我们发现，人 tankyrase 1 基因的过表达会导致裂殖酵母 Schizosaccharomyces pombe 的生长缺陷。能够恢复生长缺陷表型的化学物质可被鉴定为潜在的 tankyrase 1 抑制剂。我们使用该系统进行了高通量筛选，并鉴定出黄酮类化合物是一种能够恢复过表达 tankyrase 1 的酵母细胞生长的化合物。事实上，黄酮类化合物抑制了 tankyrase 1 过表达引起的蛋白质多聚（ADP-核糖）化。该系统可快速鉴定 tankyrase 1 的抑制活性，并且适用于使用机器人进行高通量筛选。
