A yeast one-hybrid system to detect methylation-dependent DNA-protein interactions
A yeast one-hybrid system to detect methylation-dependent DNA-protein interactions
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DOI：
10.1016/j.bbrc.2003.12.027
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发表时间：
2004-01-23
影响因子：
3.1
通讯作者：
Ito, T
中科院分区：
文献类型：
--
作者：
Feng, SY;Ota, K;Ito, T
我们开发了一种芽殖酵母基因组位点选择性CpG甲基化的方法。该方法将融合LexA的m.s sssi DNA甲基转移酶招募到目标位点附近整合的LexA操作符序列中。甲基化dna的微阵列分析表明，拴链酶选择性地甲基化目标位点周围的区域。利用这种方法在单杂交系统中甲基化诱饵DNA，我们在体内证明了甲基化依赖的DNA结合甲基化- cpg结合蛋白MBD1和Kaiso。这种依赖甲基化的单杂交系统将为寻找和分析识别甲基化DNA参与表观遗传调控的蛋白质提供一个多功能工具。(C) 2003 Elsevier Inc.版权所有。
We developed a method for site-selective CpG methylation of the budding yeast genome. The method recruits LexA-fused M.SssI DNA methyltransferase to LexA operator sequences integrated adjacent to the target site. Microarray analysis of methylated DNAs indicated that the tethered enzyme selectively methylates the region around the target site. Exploiting this method to methylate bait DNA in the one-hybrid system, we demonstrated methylation-dependent DNA binding of methyl-CpG binding proteins, MBD1 and Kaiso, in vivo. This methylation-dependent one-hybrid system would provide a versatile tool for the search and analysis of proteins that recognize methylated DNA to participate in epigenetic regulation. (C) 2003 Elsevier Inc. All rights reserved.