编码抗突变酶8-羟基鸟嘌呤-DNA糖基化酶的人cDNA的分子克隆与功能表达
Molecular cloning and functional expression of a human cDNA encoding the antimutator enzyme 8-hydroxyguanine-DNA glycosylase.
作者信息
Roldán-Arjona T, Wei Y F, Carter K C, Klungland A, Anselmino C, Wang R P, Augustus M, Lindahl T
机构信息
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire, EN6 3LD, United Kingdom.
出版信息
Proc Natl Acad Sci U S A. 1997 Jul 22;94(15):8016-20. doi: 10.1073/pnas.94.15.8016.
Abstract
The major mutagenic base lesion in DNA caused by exposure to reactive oxygen species is 8-hydroxyguanine (8-oxo-7, 8-dihydroguanine). In bacteria and Saccharomyces cerevisiae, this damaged base is excised by a DNA glycosylase with an associated lyase activity for chain cleavage. We have cloned, sequenced, and expressed a human cDNA with partial sequence homology to the relevant yeast gene. The encoded 47-kDa human enzyme releases free 8-hydroxyguanine from oxidized DNA and introduces a chain break in a double-stranded oligonucleotide specifically at an 8-hydroxyguanine residue base paired with cytosine. Expression of the human protein in a DNA repair-deficient E. coli mutM mutY strain partly suppresses its spontaneous mutator phenotype. The gene encoding the human enzyme maps to chromosome 3p25. These results show that human cells have an enzyme that can initiate base excision repair at mutagenic DNA lesions caused by active oxygen.
摘要
暴露于活性氧导致的DNA中主要的诱变碱基损伤是8-羟基鸟嘌呤（8-氧代-7,8-二氢鸟嘌呤）。在细菌和酿酒酵母中，这种受损碱基由一种具有相关裂合酶活性以进行链切割的DNA糖基化酶切除。我们已经克隆、测序并表达了一个与相关酵母基因具有部分序列同源性的人类cDNA。编码的47 kDa人类酶从氧化的DNA中释放游离的8-羟基鸟嘌呤，并在双链寡核苷酸中专门在与胞嘧啶碱基配对的8-羟基鸟嘌呤残基处引入链断裂。人类蛋白在DNA修复缺陷的大肠杆菌mutM mutY菌株中的表达部分抑制了其自发的诱变表型。编码人类酶的基因定位于染色体3p25。这些结果表明人类细胞有一种酶，它可以在由活性氧引起的诱变DNA损伤处启动碱基切除修复。
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Cloning and expression in Escherichia coli of the OGG1 gene of Saccharomyces cerevisiae, which codes for a DNA glycosylase that excises 7,8-dihydro-8-oxoguanine and 2,6-diamino-4-hydroxy-5-N-methylformamidopyrimidine.酿酒酵母OGG1基因在大肠杆菌中的克隆与表达，该基因编码一种DNA糖基化酶，可切除7,8-二氢-8-氧代鸟嘌呤和2,6-二氨基-4-羟基-5-N-甲基甲酰胺基嘧啶。
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