Recognition and reaction mechanisms of the (6-4) photolyase as determined by using a (6-4) photoproduct analog.

Junpei Yamamoto, Kenichi Hitomi, Ryosuke Hayashi, Elizabeth D Getzoff, Shigenori Iwai
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Abstract

The (6-4) photoproduct, which is one of the major UV-induced DNA lesions formed at bipyrimidine sites, causes carcinogenesis at high frequency. The (6-4) photolyases restore the (6-4) photoproducts to their intact bases in a light-dependent manner, but its overall repair mechanism remains obscure. To investigate the light-dependent conversion of the (6-4) photoproduct, we prepared a (6-4) photoproduct analog with modification at 3' pyrimidone ring, in which the carbonyl group was replaced with an imine to apply to the (6-4) photolyase assay. The (6-4) photolyase shows affinity to an oligonucleotide carrying this imine analog of the (6-4) photoproduct, though the imine analog is not repaired by the (6-4) photolyase.

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(6-4)光解酶的识别和反应机制由(6-4)光产物类似物确定。
(6-4)光产物是在联嘧啶位点形成的紫外线诱导的主要DNA损伤之一,可导致高频率的致癌。(6-4)光解酶以光依赖的方式将(6-4)光产物恢复到完整的碱基,但其整体修复机制尚不清楚。为了研究(6-4)光产物的光依赖性转化,我们制备了一个在3'嘧啶环上修饰的(6-4)光产物类似物,其中羰基被亚胺取代,用于(6-4)光解酶测定。(6-4)光解酶对携带(6-4)光产物亚胺类似物的寡核苷酸表现出亲和力,尽管亚胺类似物不能被(6-4)光解酶修复。
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