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首页> 外文期刊>Biochemistry >Mapping of peroxyl radical induced damage on genomic DNA.
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Mapping of peroxyl radical induced damage on genomic DNA.

机译：过氧自由基引发的基因组DNA损伤图谱。

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We have examined the DNA damage produced by reaction of peroxyl radicals with human fibroblast DNA. DNA damage consisted of both strand breaks and base modifications. The extent of strand breaks and base modifications induced as a function of peroxyl radical concentration was determined by quantitation of fragment size distributions using denaturing glyoxal-agarose gel electrophoresis. Both strand breaks and base modifications increased in a log linear fashion with respect to peroxyl radical concentration. Oxidative base modifications were observed to occur to a greater extent than strand breaks at every concentration measured. The sequence-specific distribution of peroxyl radical induced base damage was mapped for 803 nucleotide positions using the method of ligation mediated PCR. A total of 87% of all guanine positions in the examined sequences was found to be significantly oxidized. The order of reactivity of DNA bases toward oxidation by peroxyl radicals was found to be G C > T. Adenine is essentially unreactive. The yield of oxidative base modifications at guanines and cytosines by peroxyl radicals depends on the exact specification of 5' and 3' flanking bases in a polarity dependent manner. Every guanine in the 5'XGC3' motif was found to be oxidized, where X is any 5' neighbor. In contrast, 5' and 3' purine flanks drastically reduced the extent of peroxyl radical G oxidation. The pattern of base modification and the influence of nearest neighbors differs substantially from that previously reported for hydrogen peroxide damage mediated by low valent transition metal ions for the identical DNA sequences.
机译：我们已经检查了过氧自由基与人成纤维细胞DNA反应产生的DNA损伤。 DNA损伤包括链断裂和碱基修饰。通过使用变性乙二醛-琼脂糖凝胶电泳对片段大小分布进行定量，确定了作为过氧自由基浓度的函数而引起的链断裂和碱基修饰的程度。相对于过氧自由基浓度，链断裂和碱基修饰都以对数线性方式增加。在每种测得的浓度下，观察到氧化性碱基修饰的发生程度比链断裂更大。使用连接介导的PCR方法，将过氧自由基诱导的碱基损伤的序列特异性分布定位在803个核苷酸位置。在检查的序列中，总共87％的鸟嘌呤位置被发现被显着氧化。发现DNA碱基对过氧自由基氧化的反应顺序为G C>T。腺嘌呤基本上不反应。过氧自由基在鸟嘌呤和胞嘧啶上的氧化性碱基修饰的产率取决于极性依赖方式的5'和3'侧翼碱基的确切规格。发现5'XGC3'基序中的每个鸟嘌呤都被氧化，其中X是任何5'邻居。相反，5'和3'嘌呤侧翼大大降低了过氧自由基G的氧化程度。碱基修饰的模式和最近邻的影响与先前报道的相同DNA序列的低价过渡金属离子介导的过氧化氢损伤的模式有很大不同。

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