Graessmann M, Graessmann A, Wagner H, Werner E, Simon D
Proc Natl Acad Sci U S A. 1983 Nov;80(21):6470-4. doi: 10.1073/pnas.80.21.6470.
The effect of DNA methylation on polyoma virus and simian virus 40 gene expression was investigated. For this purpose, the cytosines of all C-G dinucleotides of the viral DNAs were methylated by the use of rat liver methylase and the completeness of methylation was verified by dinucleotide analysis and restriction endonuclease treatment. The biological activity of unmethylated and fully methylated DNAs was tested by microinjecting them into tissue culture cells. The functions analyzed included early and late viral gene expression, viral DNA replication, oncogenic transformation efficiency, and virus maturation. No difference in any of these biological functions was observed between methylated and unmethylated DNA. Early gene expression of methylated DNA is not the result of demethylation because viral DNA reextracted from the injected cells, under nonpermissive conditions, retained the methylation pattern of the input DNA. In contrast, viral DNA extracted from transformed cells or from intact virus particles was partially or completely demethylated.
研究了DNA甲基化对多瘤病毒和猴病毒40基因表达的影响。为此,使用大鼠肝脏甲基化酶对病毒DNA的所有C-G二核苷酸的胞嘧啶进行甲基化,并通过二核苷酸分析和限制性内切酶处理验证甲基化的完整性。通过将未甲基化和完全甲基化的DNA显微注射到组织培养细胞中来测试其生物活性。分析的功能包括病毒早期和晚期基因表达、病毒DNA复制、致癌转化效率和病毒成熟。甲基化和未甲基化的DNA在这些生物学功能上均未观察到差异。甲基化DNA的早期基因表达不是去甲基化的结果,因为在非允许条件下从注射细胞中重新提取的病毒DNA保留了输入DNA的甲基化模式。相反,从转化细胞或完整病毒颗粒中提取的病毒DNA部分或完全去甲基化。