小鼠DNA中N6-甲基腺嘌呤水平不可检测：PRED28的克隆与分析，PRED28是一个编码假定哺乳动物DNA腺嘌呤甲基转移酶的基因。

Undetectable levels of N6-methyl adenine in mouse DNA: Cloning and analysis of PRED28, a gene coding for a putative mammalian DNA adenine methyltransferase.

作者信息

Ratel David, Ravanat Jean-Luc, Charles Marie-Pierre, Platet Nadine, Breuillaud Lionel, Lunardi Joël, Berger François, Wion Didier

机构信息

INSERM U318, UJFG, CHU Michallon, 38043 Grenoble, France.

出版信息

FEBS Lett. 2006 May 29;580(13):3179-84. doi: 10.1016/j.febslet.2006.04.074. Epub 2006 May 3.

DOI:10.1016/j.febslet.2006.04.074

PMID:16684535

Abstract

Three methylated bases, 5-methylcytosine, N4-methylcytosine and N6-methyladenine (m6A), can be found in DNA. However, to date, only 5-methylcytosine has been detected in mammalian genomes. To reinvestigate the presence of m6A in mammalian DNA, we used a highly sensitive method capable of detecting one N6-methyldeoxyadenosine per million nucleosides. Our results suggest that the total mouse genome contains, if any, less than 10(3) m6A. Experiments were next performed on PRED28, a putative mammalian N6-DNA methyltransferase. The murine PRED28 encodes two alternatively spliced RNA. However, although recombinant PRED28 proteins are found in the nucleus, no evidence for an adenine-methyltransferase activity was detected.

摘要

DNA中可发现三种甲基化碱基，即5-甲基胞嘧啶、N4-甲基胞嘧啶和N6-甲基腺嘌呤（m6A）。然而，迄今为止，在哺乳动物基因组中仅检测到5-甲基胞嘧啶。为了重新研究m6A在哺乳动物DNA中的存在情况，我们使用了一种高度灵敏的方法，该方法能够检测每百万个核苷中的一个N6-甲基脱氧腺苷。我们的结果表明，小鼠基因组中即使存在m6A，其含量也少于10³个。接下来对一种假定的哺乳动物N6-DNA甲基转移酶PRED28进行了实验。小鼠PRED28编码两种可变剪接RNA。然而，尽管在细胞核中发现了重组PRED28蛋白，但未检测到腺嘌呤甲基转移酶活性的证据。