Chen J, Derfler B, Samson L
Laboratory of Toxicology, Harvard School of Public Health, Boston, MA 02115.
EMBO J. 1990 Dec;9(13):4569-75. doi: 10.1002/j.1460-2075.1990.tb07910.x.
We previously cloned a DNA fragment from Saccharomyces cerevisiae that suppressed the alkylation sensitivity of Escherichia coli glycosylase deficient mutants and we showed that it apparently contained a gene for 3-methyl-adenine DNA glycosylase (MAG). Here we establish the identity of the MAG gene by sequence analysis and describe its in vivo function and expression in yeast cells. The MAG DNA glycosylase specifically protects yeast cells against the killing effects of alkylating agents. It does not protect cells against mutation; indeed, it appears to generate mutations which presumably result from those apurinic sites produced by the glycosylase that escape further repair. The MAG gene, which we mapped to chromosome V, is not allelic with any of the RAD genes and appears to be allelic to the unmapped MMS-5 gene. From its sequence the MAG glycosylase is predicted to contain 296 amino acids and have a molecular weight of 34,293 daltons. A 137 amino acid stretch of the MAG glycosylase displays 27.0% identity and 63.5% similarity with the E. coli AlkA glycosylase. Transcription of the MAG gene, like that of the E. coli alkA gene, is greatly increased when yeast cells are exposed to relatively non-toxic levels of alkylating agents.
我们之前从酿酒酵母中克隆了一个DNA片段,该片段可抑制大肠杆菌糖基化酶缺陷型突变体对烷基化的敏感性,并且我们发现它显然包含一个3 - 甲基腺嘌呤DNA糖基化酶(MAG)基因。在此,我们通过序列分析确定了MAG基因的身份,并描述了其在酵母细胞中的体内功能和表达情况。MAG DNA糖基化酶可特异性保护酵母细胞免受烷基化剂的杀伤作用。它不能保护细胞免受突变影响;实际上,它似乎会产生突变,这些突变大概是由糖基化酶产生的那些未进一步修复的脱嘌呤位点导致的。我们将MAG基因定位到了第五条染色体上,它与任何RAD基因都不等位,似乎与未定位的MMS - 5基因等位。根据其序列预测,MAG糖基化酶含有296个氨基酸,分子量为34293道尔顿。MAG糖基化酶的一段137个氨基酸的序列与大肠杆菌AlkA糖基化酶有27.0%的同一性和63.5%的相似性。当酵母细胞暴露于相对无毒水平的烷基化剂时,MAG基因的转录,就像大肠杆菌alkA基因的转录一样,会大幅增加。