来自再生大鼠肝脏的DNA甲基化酶：纯化与特性鉴定
DNA-methylase from regenerating rat liver: purification and characterisation.
作者信息
Simon D, Grunert F, von Acken U, Döring H P, Kröger H
出版信息
Nucleic Acids Res. 1978 Jun;5(6):2153-67. doi: 10.1093/nar/5.6.2153.
Abstract
DNA methylase has been purified 660-fold from nuclei from regenerating rat liver. The enzyme is able to methylate single stranded (ss) and double stranded (ds) DNA, the only reaction product being 5-methylcytosine. Previously unmethylated double stranded DNA from prokaryotes (M.luteus) as well as from eukaryotes (Ascaris suis) can serve as substrates. The synthetic copolymers (dG-dC)n . (dC-dG)n and (dG,dC)n are also methylated. While SV40 DNA is almost not methylated, PM2 DNA is a good substrate even in the supercoiled form. The enzyme methylates 1 in 17 bases in heterologous M.luteus DNA, but only 1 in 590 in homologous rat liver DNA. The high methylation level of M.luteus DNA, an analysis of the methylated pyrimidine isostichs and a preliminary dinucleotide analysis suggest that all the CpGs in a DNA can be methylated.
摘要
已从再生大鼠肝脏的细胞核中纯化出DNA甲基化酶，纯化倍数达660倍。该酶能够使单链（ss）和双链（ds）DNA甲基化，唯一的反应产物是5-甲基胞嘧啶。来自原核生物（藤黄微球菌）以及真核生物（猪蛔虫）的先前未甲基化的双链DNA都可以作为底物。合成共聚物（dG-dC）n·（dC-dG）n和（dG，dC）n也能被甲基化。虽然SV40 DNA几乎不被甲基化，但PM2 DNA即使处于超螺旋形式也是良好的底物。该酶使异源的藤黄微球菌DNA中17个碱基中的1个甲基化，但在同源的大鼠肝脏DNA中仅使590个碱基中的1个甲基化。藤黄微球菌DNA的高甲基化水平、对甲基化嘧啶等距序列的分析以及初步的二核苷酸分析表明，DNA中的所有CpG都可以被甲基化。
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