Reversal of H3K9me2 by a small-molecule inhibitor for the G9a histone methyltransferase
Reversal of H3K9me2 by a small-molecule inhibitor for the G9a histone methyltransferase
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DOI：
10.1016/j.molcel.2007.01.017
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发表时间：
2007-02-09
期刊：
影响因子：
16
通讯作者：
Jenuwein, Thomas
中科院分区：
文献类型：
--
作者：
Kubicek, Stefan;O'Sullivan, Roderick J.;Jenuwein, Thomas
组蛋白赖氨酸甲基化在染色质结构域的组织和基因表达的调节中具有重要作用。为了分析其功能并调节其活性，我们使用重组G9A作为靶酶筛选了针对组蛋白赖氨酸甲基转移酶（HMTase）的特异性抑制剂。从包含125,000种预选化合物的化学库中，发现了7次命中。其中一种抑制剂BIX-01294（地氮杂素唑胺 - 胺衍生物）不会与辅因子S-腺苷甲硫代氨酸竞争，并选择性地损害了G9A HMTase和H3K9me2的产生。在细胞测定中，与Bix-01 294的几个细胞系的瞬时孵育可在去除抑制剂后恢复的批量H3K9me2水平。重要的是，几个G9A靶基因的染色质免疫沉淀表明在抑制剂治疗的小鼠ES细胞和成纤维细胞中，启动子良性H3K9me2的可逆降低。我们的数据确定了具有生物活性的HMTase抑制剂，该抑制剂允许在哺乳动物染色质中进行H3K9ME2标记的瞬时调节。
Histone lysine methylation has important roles in the organization of chromatin domains and the regulation of gene expression. To analyze its function and modulate its activity, we screened for specific inhibitors against histone lysine methyltransferases (HMTases) using recombinant G9a as the target enzyme. From a chemical library comprising 125,000 preselected compounds, seven hits were identified. Of those, one inhibitor, BIX-01294 (diazepinquinazolin-amine derivative), does not compete with the cofactor S-adenosyl-methionine, and selectively impairs the G9a HMTase and the generation of H3K9me2 in vitro. In cellular assays, transient incubation of several cell lines with BIX-01 294 lowers bulk H3K9me2 levels that are restored upon removal of the inhibitor. Importantly, chromatin immunoprecipitation at several G9a target genes demonstrates reversible reduction of promoter-proximal H3K9me2 in inhibitor-treated mouse ES cells and fibroblasts. Our data identify a biologically active HMTase inhibitor that allows for the transient modulation of H3K9me2 marks in mammalian chromatin.