SuaI限制修饰系统介导的胞嘧啶甲基化：对嗜热古菌遗传保真度的影响
Cytosine methylation by the SuaI restriction-modification system: implications for genetic fidelity in a hyperthermophilic archaeon.
作者信息
Grogan Dennis W
机构信息
New England Biolabs, Inc., Beverly, Massachusetts 01915, USA.
出版信息
J Bacteriol. 2003 Aug;185(15):4657-61. doi: 10.1128/JB.185.15.4657-4661.2003.
Abstract
5-methylcytosine in chromosomal DNA represents a potential source of frequent spontaneous mutation for hyperthermophiles. To determine the relevance of this threat for the archaeon Sulfolobus acidocaldarius, the mode of GGCC methylation by its restriction-modification system, SuaI, was investigated. Distinct isoschizomers of the SuaI endonuclease were used to probe the methylation state of GGCC in native S. acidocaldarius DNA. In addition, the methylation sensitivity of the SuaI endonuclease was determined with synthetic oligonucleotide substrates and modified natural DNAs. The results show that the SuaI system uses N(4) methylation to block cleavage of its recognition site, thereby avoiding the creation of G. T mismatches by spontaneous deamination at extremely high temperature.
摘要
染色体DNA中的5-甲基胞嘧啶是嗜热菌频繁自发突变的潜在来源。为了确定这种威胁对嗜热栖热放线菌的相关性，研究了其限制修饰系统SuaI对GGCC的甲基化模式。使用SuaI内切核酸酶的不同同裂酶来探测天然嗜热栖热放线菌DNA中GGCC的甲基化状态。此外，用合成寡核苷酸底物和修饰的天然DNA测定了SuaI内切核酸酶的甲基化敏感性。结果表明，SuaI系统利用N(4)甲基化来阻断其识别位点的切割，从而避免在极高温度下通过自发脱氨产生G.T错配。
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