On the mechanism of demethylation of 5-methylcytosine in DNA

被引：44

作者：

Hamm, Stefan ^{[3
]}

Just, George ^{[3
]}

Lacoste, Nathalie ^{[3
]}

Moitessier, Nicolas ^{[3
]}

Szyf, Moshe ^{[2
]}

Mamer, Orval ^{[1
]}

机构：

[1] McGill Univ, Mass Spectrometry Unit, Montreal, PQ H3A 1A4, Canada

[2] McGill Univ, Dept Pharmacol & Therapeut, Montreal, PQ H3A 1A4, Canada

[3] McGill Univ, Dept Chem, Montreal, PQ, Canada

来源：

BIOORGANIC & MEDICINAL CHEMISTRY LETTERS | 2008年 / 18卷 / 03期

基金：

加拿大健康研究院;

关键词：

gas chromatography-mass spectrometry; DNA; methylcytosine; demethylation; DNA demethylation;

D O I：

10.1016/j.bmcl.2007.12.027

中图分类号：

R914 [药物化学];

学科分类号：

100701 ;

摘要：

DNA methylation is an important biological process that programmes gene expression in vertebrates. The methylation pattern is generated by a combination of methylation and demethylation reactions catalyzed by DNA methyltransferases and putative demethylases. MBD2 binds methylated DNA and possesses DNA demethylase activity. We use here direct analysis of the reaction mixture by GC-MS using a water-tolerant gas chromatographic column to avoid the loss of potential volatile products and identify the leaving residue of the demethylation reaction. We show that the DNA demethylase reaction catalyzed by a recombinant human MBD2 purified from SF9 insect cells releases dideuteroformaldehyde from [Me-H-2(3)]-5-methylcytosine in DNA. A mechanism of the DNA demethylation reaction is proposed based on this observation. (C) 2007 Elsevier Ltd. All rights reserved.

引用

页码：1046 / 1049

页数：4

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