Characterization of Undermethylated Sites in Vibrio cholerae
Characterization of Undermethylated Sites in Vibrio cholerae
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DOI：
10.1128/jb.02112-12
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发表时间：
2013-05-01
影响因子：
3.2
通讯作者：
Camilli, Andrew
中科院分区：
文献类型：
--
作者：
Dalia, Ankur B.;Lazinski, David W.;Camilli, Andrew
DNA甲基转移酶的活性对于细菌中的多种细胞功能是重要的。在这项研究中，我们开发了一种改进的高通量技术，称为甲基均聚物尾介导的测序（甲基HTM-seq），以确定在霍乱弧菌基因组中的两个DNA甲基转移酶，大坝，腺嘌呤甲基转移酶，和VchM，胞嘧啶甲基转移酶，在丰富的培养基在体外生长的甲基化不足的网站。许多甲基化不足的位点发生在基因间区域，对于大多数这些位点，我们确定了负责甲基化不足的转录因子。这证实了先前假设的这些转录因子的DNA-蛋白质相互作用的存在，并提供了对这些细胞在体外生长过程中的生物学状态的了解。此前已证明DNA腺嘌呤甲基化介导基因调节中的遗传表观遗传开关。然而，没有一个未充分甲基化的大坝网站测试表明，这种机制的调节证据。这项研究是第一个使用第二代测序技术在细菌全基因组范围内鉴定甲基化不足的腺嘌呤和胞嘧啶。
The activities of DNA methyltransferases are important for a variety of cellular functions in bacteria. In this study, we developed a modified high-throughput technique called methyl homopolymer tail mediated sequencing (methyl HTM-seq) to identify the undermethylated sites in the Vibrio cholerae genome for the two DNA methyltransferases, Dam, an adenine methyltransferase, and VchM, a cytosine methyltransferase, during growth in rich medium in vitro. Many of the undermethylated sites occurred in intergenic regions, and for most of these sites, we identified the transcription factors responsible for undermethylation. This confirmed the presence of previously hypothesized DNA-protein interactions for these transcription factors and provided insight into the biological state of these cells during growth in vitro. DNA adenine methylation has previously been shown to mediate heritable epigenetic switches in gene regulation. However, none of the undermethylated Dam sites tested showed evidence of regulation by this mechanism. This study is the first to identify undermethylated adenines and cytosines genomewide in a bacterium using second-generation sequencing technology.