Novel small-molecule inhibitors of RNA polymerase III
Novel small-molecule inhibitors of RNA polymerase III
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DOI：
10.1128/ec.2.2.256-264.2003
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发表时间：
2003-04-01
期刊：
影响因子：
--
通讯作者：
McGovern, K
中科院分区：
文献类型：
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作者：
Wu, LP;Pan, J;McGovern, K
利用酿酒酵母（Saccharomyces cerevisiae）的一种遗传学方法被用于鉴定抗真菌化合物的靶点。这种分析导致了对酿酒酵母RNA聚合酶（Pol）III小分子抑制剂的鉴定。三条证据表明UK - 118005通过靶向酵母中的RNA Pol III抑制细胞生长。首先，RNA Pol III最大亚基Rpo31p的g结构域的一个显性突变赋予了对该化合物的抗性。其次，UK - 118005在野生型细胞中迅速抑制tRNA合成，但在UK - 118005抗性突变体中则不抑制。第三，在生化分析中，UK - 118005在体外抑制野生型而非突变型Pol III酶的tRNA基因转录。通过在一种模板特异性RNA Pol III转录分析中测试UK - 118005的类似物，鉴定出一种效力显著更高的抑制剂ML - 60218。进一步的检测表明这两种化合物都是广谱抑制剂，对源自白色念珠菌和人类细胞的RNA Pol III转录系统显示出活性。这些抑制剂的鉴定表明RNA Pol III可被小分子合成物作为靶点。
A genetic approach utilizing the yeast Saccharomyces cerevisiae was used to identify the target of antifungal compounds. This analysis led to the identification of small molecule inhibitors of RNA polymerase (Poll) III from Saccharomyces cerevisiae. Three lines of evidence show that UK-118005 inhibits cell growth by targeting RNA Poll III in yeast. First, a dominant mutation in the g domain of Rpo31p, the largest subunit of RNA Pol III, confers resistance to the compound. Second, UK-118005 rapidly inhibits tRNA synthesis in wild-type cells but not in UK-118005 resistant mutants. Third, in biochemical assays, UK-118005 inhibits tRNA gene transcription in vitro by the wild-type but not the mutant Pol III enzyme. By testing analogs of UK-118005 in a template-specific RNA Poll III transcription assay, an inhibitor with significantly higher potency, ML-60218, was identified. Further examination showed that both compounds are broad-spectrum inhibitors, displaying activity against RNA Pol III transcription systems derived from Candida albicans and human cells. The identification of these inhibitors demonstrates that RNA Pol III can be targeted by small synthetic molecules.