Sohail A, Lieb M, Dar M, Bhagwat A S
Cold Spring Harbor Laboratory, New York 11724.
J Bacteriol. 1990 Aug;172(8):4214-21. doi: 10.1128/jb.172.8.4214-4221.1990.
Deamination of 5-methylcytosine in DNA results in T/G mismatches. If unrepaired, these mismatches can lead to C-to-T transition mutations. The very short patch (VSP) repair process in Escherichia coli counteracts the mutagenic process by repairing the mismatches in favor of the G-containing strand. Previously we have shown that a plasmid containing an 11-kilobase fragment from the E. coli chromosome can complement a chromosomal mutation defective in both cytosine methylation and VSP repair. We have now mapped the regions essential for the two phenotypes. In the process, we have constructed plasmids that complement the chromosomal mutation for methylation, but not for repair, and vice versa. The genes responsible for these phenotypes have been identified by DNA sequence analysis. The gene essential for cytosine methylation, dcm, is predicted to code for a 473-amino-acid protein and is not required for VSP repair. It is similar to other DNA cytosine methylases and shares extensive sequence similarity with its isoschizomer, EcoRII methylase. The segment of DNA essential for VSP repair contains a gene that should code for a 156-amino-acid protein. This gene, named vsr, is not essential for DNA methylation. Remarkably, the 5' end of this gene appears to overlap the 3' end of dcm. The two genes appear to be transcribed from a common promoter but are in different translational registers. This gene arrangement may assure that Vsr is produced along with Dcm and may minimize the mutagenic effects of cytosine methylation.
DNA中5 - 甲基胞嘧啶的脱氨基作用会导致T/G错配。如果不进行修复,这些错配会导致C到T的转换突变。大肠杆菌中的极短补丁(VSP)修复过程通过修复有利于含G链的错配来对抗诱变过程。此前我们已经表明,一个含有来自大肠杆菌染色体11千碱基片段的质粒可以弥补在胞嘧啶甲基化和VSP修复方面都有缺陷的染色体突变。我们现在已经绘制出了这两种表型所必需的区域。在此过程中,我们构建了能够弥补染色体甲基化突变但不能弥补修复突变的质粒,反之亦然。通过DNA序列分析确定了负责这些表型的基因。胞嘧啶甲基化所必需的基因dcm预计编码一个473个氨基酸的蛋白质,并且VSP修复不需要它。它与其他DNA胞嘧啶甲基转移酶相似,并且与其同裂酶EcoRII甲基转移酶具有广泛的序列相似性。VSP修复所必需的DNA片段包含一个应该编码156个氨基酸蛋白质的基因。这个基因名为vsr,对DNA甲基化不是必需的。值得注意的是,这个基因的5'端似乎与dcm的3'端重叠。这两个基因似乎从一个共同的启动子转录,但处于不同的翻译阅读框。这种基因排列可能确保Vsr与Dcm一起产生,并可能将胞嘧啶甲基化的诱变作用降至最低。