Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account.
Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account.
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DOI：
10.2183/pjab.84.321
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发表时间：
2008
期刊：
影响因子：
--
通讯作者：
Hayatsu H
中科院分区：
文献类型：
--
作者：
Hayatsu H
DNA中胞嘧啶5位甲基化与基因功能的调控密切相关，因而在生物科学的许多领域都得到了广泛的研究。在序列水平上确定5-甲基胞嘧啶在基因组中的位置的主要分析方法是对DNA进行亚硫酸氢盐修饰。这种化学处理的用途是基于亚硫酸氢盐选择性脱氨基胞嘧啶残基的特性。亚硫酸氢盐介导的胞嘧啶脱氨反应是我们于1970年在东京大学发现的。与此同时，夏皮罗和他在纽约大学的同事独立地发现了同样的反应。我们还报道了亚硫酸氢盐对5-甲基胞嘧啶的脱氨反应非常缓慢。这些发现后来被一组澳大利亚科学家用来设计一种分析DNA中5-甲基胞嘧啶的方法；因此，这些研究人员在1992年发明了一种名为亚硫酸氢盐基因组测序的方法。这篇综述描述了作者对发现亚硫酸盐与嘧啶碱反应的思考。本文还叙述了作者最近用一种新设计的高浓度亚硫酸氢盐溶液改进分析程序的工作。
Methylation at position 5 of cytosine in DNA is being intensively studied in many areas of biological sciences, as the methylation is intimately associated with the control of gene functions. The principal analytical method for determining the sites of 5-methylcytosine in genome at the sequence level involves bisulfite modification of DNA. The utility of this chemical treatment is based on the property of bisulfite to selectively deaminate cytosine residues. The bisulfite-mediated cytosine deamination was discovered in 1970 by us in the University of Tokyo. At the same time, Shapiro and his coworkers in New York University found the same reaction independently. We also reported that 5-methylcytosine was deaminated by bisulfite only very slowly. These findings were later utilized by a group of Australian scientists to devise a means to analyze 5-methylcytosine in DNA; thus, a method called ‘bisulfite genomic sequencing’ was invented by these researchers in 1992. This review describes the author’s reflection of the discovery of bisulfite reactions with pyrimidine bases. The author’s recent work that has resulted in an improvement of the procedure of analysis by use of a newly devised high concentration bisulfite solution is also described.
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