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首页> 外文期刊>Biochemistry >In vitro replication and repair of DNA containing a C2'-oxidized abasic site.
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In vitro replication and repair of DNA containing a C2'-oxidized abasic site.

机译：体外复制和修复含有C2'-氧化脱脂位点的DNA。

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摘要

Abasic lesions are unable to form Watson-Crick hydrogen bonds with nucleotides. Nonetheless, polymerase and repair enzymes distinguish between various oxidized abasic lesions, as well as from nonoxidized abasic sites (AP). The C2-AP lesion is produced when DNA is exposed to gamma-radiolysis. Its effects on polymerases and repair enzymes are unknown. A recently reported method for the chemical synthesis of oligonucleotides containing C2-AP at a defined site was utilized for studying the activity of Klenow exo(-) and repair enzymes on templates containing the lesion. The C2-AP lesion has a similar effect on Klenow exo(-) as do AP and C4-AP sites. Deoxyadenosine is preferentially incorporated opposite C2-AP, but extension of the primer past the lesion is strongly blocked. C2-AP is incised less efficiently by exonuclease III and endonuclease IV than are other abasic lesions. Furthermore, although a Schiff base between C2-AP and endonuclease III can be chemically trapped, the location of the 3'-phosphate alpha with respect to the aldehyde prevents beta-elimination associated with the lyase activity of type I base excision repair enzymes. The interactions of the C2'-oxidized abasic site with Klenow exo(-) and repair enzymes suggest that the lesion will be mutagenic and that it will be removed by strand displacement synthesis and flap endonuclease processing via a long patch repair mechanism.
机译：脱脂病变不能与核苷酸形成Watson-Crick氢键。尽管如此，聚合酶和修复酶区分各种氧化的饮料病变，以及非氧化释放位点（AP）。当DNA暴露于γ-辐射溶解时，产生C2-AP病变。它对聚合酶和修复酶的影响是未知的。最近报道了含有在限定位点的C2-AP的寡核苷酸化学合成方法，用于研究Klenow Exo（ - ）和修复酶的含有病变的模板的活性。 C2-AP病变对Klenow Exo（ - ）和C4-AP位点具有类似的效果。脱氧腺苷优先掺入C2-AP相对，但底漆过底漆过损伤的延伸强烈堵塞。通过外切核酸酶III和内切核酸酶IV，C2-AP效率较低，而不是其他易行的病变。此外，尽管C2-AP和内​​切核酸酶III之间的Schiff基碱可以进行化学捕获，但是3'-磷酸α相对于醛的位置可防止与I型基本切除修复酶的裂解酶活性相关的β-消除。 C2'-氧化的脱水位点与Klenow EXO（ - - ）和修复酶的相互作用表明病变将是致突变性的，并且通过长贴片修复机制将通过链置换合成和皮瓣内切核酸酶加工除去。

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