Methylglyoxal, an endogenous, aldehyde, crosslinks DNA polymerase and the substrate DNA
Methylglyoxal, an endogenous, aldehyde, crosslinks DNA polymerase and the substrate DNA
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DOI：
10.1093/nar/29.16.3433
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发表时间：
2001-08-15
影响因子：
14.9
通讯作者：
Kamiya, H
中科院分区：
文献类型：
--
作者：
Murata-Kamiya, N;Kamiya, H
丙酮醛是一种已知的内源性和环境诱变剂，是一种反应性α-酮醛，可以修饰DNA和蛋白质。为了研究甲基乙二醛诱导DNA和DNA聚合酶之间交联的可能性，我们在体外用甲基乙二醛处理了“引物模板”DNA和DNA聚合酶I的核酸外切酶缺陷型Klenow片段（KFexo-）。当通过SDS-PAGE分析反应混合物时，我们发现甲基乙二醛诱导DNA-KFex-交联。KFexo-和“引物模板”DNA的特异性结合复合物对于DNA-KFexo-交联的形成是必需的。甲基乙二醛与模板DNA的单链部分中的鸟嘌呤残基反应。当2 '-脱氧鸟苷与N-α-乙酰赖氨酸或N-乙酰半胱氨酸在甲基乙二醛存在下孵育时，形成交联产物。没有其他测试的氨基酸衍生物可以产生交联产物。这些结果表明，甲基乙二醛交联的底物DNA的鸟嘌呤残基和赖氨酸和半胱氨酸残基附近的DNA聚合酶的结合位点在DNA合成过程中，DNA复制受到严重抑制的甲基乙二醛诱导的DNA-DNA聚合酶交联。
Methylglyoxal, a known endogenous and environmental mutagen, is a reactive alpha -ketoaldehyde that can modify both DNA and proteins. To investigate the possibility that methylglyoxal induces a crosslink between DNA and DNA polymerase, we treated a 'primed template' DNA and the exonuclease-deficient Klenow fragment (KFexo-) of DNA polymerase I with methylglyoxal in vitro. When the reaction mixtures were analyzed by SDS-PAGE, we found that methylglyoxal induced a DNA-KFexo- crosslink. The specific binding complex of KFexo- and 'primed template' DNA was necessary for formation of the DNA-KFexo- crosslink. Methylglyoxal reacted with guanine residues in the single-stranded portion of the template DNA. When 2'-deoxyguanosine was incubated with N alpha -acetyllysine or N-acetylcysteine in the presence of methylglyoxal, a crosslinked product was formed. No other amino acid derivatives tested could generate a crosslinked product. These results suggest that methylglyoxal crosslinks a guanine residue of the substrate DNA and lysine and cysteine residues near the binding site of the DNA polymerase during DNA synthesis and that DNA replication is severely inhibited by the methylglyoxal-induced DNA-DNA polymerase crosslink.