Kudriashova I B, Kirnos M D, Vaniushin B F
Biokhimiia. 1976 Nov;41(11):1968-77.
DNA-methylase activities which methylate cytosine residues in homo- and heterologous DNA were detected in mitochondria and nuclei from rat liver and beef heart. Adenine modifying DNA-methylases in mitochondria and nuclei were not found. DNA from mitochondria and nuclei differ significantly in the methylation degree and in the pattern of the 5-methyl-cytosine distribution by pyrimidine isostichs as DNA in vivo and in vitro being methylated. Mitochondrial DNA methylase has the maximum activity at 30 degrees and pH 7.8 this enzyme(s) differ(s) from the nuclear one(s) in the pH dependence of its activity. After exhaustive in vitro methylation of various DNA by the nuclear enzyme DNA-methylase from mitochondria additionally introduces CH3 groups from S-adenosylmethionine into these DNA (about 3 times more CH3 groups than nuclear enzyme). Nuclear DNA-methylase also methylates DNA which is previously fully-methylated by the mitochondrial enzyme, but to a lesser degree. In conditions of exhaustive DNA methylation mitochondrial enzyme introduces into E. coli B DNA about four times more methyl groups as compared to the nuclear one. After the methylation of E. coli B DNA by mitochondrial enzyme the label (3H-methyl) was detected predominantly in mono-, and in case of nuclear enzyme--in di- and tripyrimidine fragments. Mitochondrial DNA-methylase differs from the nuclear one in the nature of recognized DNA sequences; these enzymes seems to be represented by different proteins. The mitochondrial enzyme methylates shorter nucleotide sequences in DNA as compared to the nuclear DNA-methylase. All these data suggest there exist organoid specificity of genome methylation in animal cell and the modification-restriction systems in animal nucleus and mitochondria are different in character.
在大鼠肝脏和牛心脏的线粒体及细胞核中,检测到了能使同源和异源DNA中的胞嘧啶残基发生甲基化的DNA甲基化酶活性。未发现线粒体和细胞核中存在使腺嘌呤发生修饰的DNA甲基化酶。线粒体和细胞核中的DNA在甲基化程度以及5 - 甲基胞嘧啶通过嘧啶同列线分布的模式上存在显著差异,因为体内和体外的DNA都会发生甲基化。线粒体DNA甲基化酶在30℃和pH 7.8时具有最大活性,该酶在活性的pH依赖性方面与核内的酶不同。在用核酶对各种DNA进行彻底的体外甲基化后,线粒体中的DNA甲基化酶还能将S - 腺苷甲硫氨酸中的CH₃基团引入这些DNA中(引入的CH₃基团比核酶多约3倍)。核DNA甲基化酶也能使先前已被线粒体酶完全甲基化的DNA发生甲基化,但程度较低。在DNA彻底甲基化的条件下,线粒体酶引入到大肠杆菌B DNA中的甲基基团数量是核酶的约四倍。用线粒体酶对大肠杆菌B DNA进行甲基化后,标记(³H - 甲基)主要在单嘧啶片段中被检测到,而用核酶进行甲基化时,则在双嘧啶和三嘧啶片段中被检测到。线粒体DNA甲基化酶在识别的DNA序列性质上与核内的酶不同;这些酶似乎由不同的蛋白质组成。与核DNA甲基化酶相比,线粒体酶使DNA中较短的核苷酸序列发生甲基化。所有这些数据表明动物细胞中存在基因组甲基化的细胞器特异性,并且动物细胞核和线粒体中的修饰 - 限制系统在性质上是不同的。