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首页> 外文期刊>Biochemistry >In vitro reconstitution of the Schizosaccharomyces pombe alternative excision repair pathway.
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In vitro reconstitution of the Schizosaccharomyces pombe alternative excision repair pathway.

机译：粟酒裂殖酵母的体外重建替代切除修复途径。

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摘要

Schizosaccharomyces pombe alternative excision repair has been shown genetically and biochemically to be involved in the repair of a wide variety of DNA lesions. AER is initiated by a damage-specific endonuclease (Uve1p) that recognizes UV-induced photoproducts, base mispairs, abasic sites, and platinum G-G diadducts and cleaves the DNA phosphodiester backbone 5' to a lesion. Several models exist that employ various mechanisms for damage removal based on the activities of Rad2p, a nuclease thought to be responsible for damage excision in AER. This study represents the first report of the biochemical reconstitution of the AER pathway. A base mispair-containing substrate is repaired in a reaction requiring S. pombe Uve1p, Rad2p, DNA polymerase delta, replication factor C, proliferating cell nuclear antigen, and T4 DNA ligase. Surprisingly, damage is removed exclusively by the 5' to 3' exonuclease activity of Rad2p and not its "flap endonuclease" activity and is absolutely dependent upon the presence of the 5'-phosphoryl moiety at the Uve1p cleavage site.
机译：粟酒裂殖酵母的替代性切除修复已在遗传和生物化学方面显示出与多种DNA损伤的修复有关。 AER由损伤特异性核酸内切酶（Uve1p）启动，该酶识别UV诱导的光产物，碱基错配，无碱基位点和铂G-G二加合物，并将DNA磷酸二酯主链5'裂解为病变。存在几种基于Rad2p活性采用多种机制消除损伤的模型，Rad2p是一种核酸酶，被认为是造成AER中损伤切除的原因。这项研究代表了AER途径的生化重构的首次报道。在需要粟酒裂殖酵母Uve1p，Rad2p，DNA聚合酶δ，复制因子C，增殖细胞核抗原和T4 DNA连接酶的反应中，修复了含有碱基配对错误的底物。出人意料的是，仅通过Rad2p的5'至3'核酸外切酶活性而不是其“瓣内切核酸酶”活性来清除损伤，并且完全取决于Uve1p切割位点上5'-磷酰基部分的存在。

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