Mushkambarov N N, Votrin I I, Debov S S
Biokhimiia. 1978 Jun;43(6):1111-20.
Evidence from comparative determination of DNA radioactivity methylation degree of acidic extraction and chlorophormic deproteination of the samples suggest that the former technique is a more efficient one. The properties of the DNA-methylase reaction in isolated rat liver nuclei were studied. The DNA-methylase activity is found to be considerably stable during incubation of the nuclei at 37 degrees C; a broad pH-optimum in the alkaline region is observed (pH 8.6--9.8); this activity is inhibited by Mn2+, nucleotides, actynomycin and S-adenosyl methionine analogs and is activated by Mg2+; the incorporation of methyl groups into DNA is reversible. The data suggest that the DNA-methylase activities of the nuclei isolated at different stages of regeneration do not show substantial variations. No differences in DNA methylation before and after DNA synthesis in the regenerating nuclei were observed. Inhibition of DNA synthesis in the course of regeneration does not decrease the level of DNA methylation. The interrelationship between methylation and replication of DNA is discussed.
对样品进行酸性提取和氯仿脱蛋白后DNA放射性甲基化程度的比较测定结果表明,前一种技术效率更高。研究了分离的大鼠肝细胞核中DNA甲基化酶反应的特性。发现DNA甲基化酶活性在细胞核于37℃孵育期间相当稳定;在碱性区域观察到较宽的pH最适范围(pH 8.6 - 9.8);该活性受到Mn2+、核苷酸、放线菌素和S-腺苷甲硫氨酸类似物的抑制,并被Mg2+激活;甲基基团掺入DNA是可逆的。数据表明,在再生不同阶段分离的细胞核的DNA甲基化酶活性没有显著变化。在再生细胞核中,DNA合成前后的DNA甲基化没有差异。再生过程中DNA合成的抑制不会降低DNA甲基化水平。讨论了DNA甲基化与复制之间的相互关系。