Proliferating cell nuclear antigen-dependent abasic site repair in Xenopus laevis oocytes: an alternative pathway of base excision DNA repair.
Proliferating cell nuclear antigen-dependent abasic site repair in Xenopus laevis oocytes: an alternative pathway of base excision DNA repair.
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爪蟾卵母细胞增殖细胞核抗原依赖性脱碱基位点修复：碱基切除 DNA 修复的替代途径。
DOI：
10.1128/mcb.14.9.6187-6197.1994
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发表时间：
1994
影响因子：
5.3
通讯作者：
Bogenhagen,DF
中科院分区：
文献类型：
--
作者：
Matsumoto,Y;Kim,K;Bogenhagen,DF
DNA损伤经常导致apurinic/ ap嘧啶（AP）位点的产生，这些位点被认为是通过碱基切除途径修复的。为了详细分析这种修复机制，在模型系统中使用了AP位点的合成类似物3-羟基-2-羟甲基四氢呋喃（tetrahydrofuran）。四氢呋喃残基在青蛇卵细胞提取物中被有效修复，其中大多数修复事件涉及不超过四个核苷酸的atp依赖性结合（Y. Matsumoto和D. F. Bogenhagen, Mol. Cell）。9:3750-3757, 1989；Y. Matsumoto和D. F. Bogenhagen, Mol Cell。《圣经》11:44 . 41 - 44 . 47,1991)。使用一系列的柱层析程序来分馏atex。我们开发了一个重组的四氢呋喃修复系统，包括五个部分，其中三个部分纯化到接近均匀：增殖细胞核抗原（PCNA）， AP内切酶和DNA聚合酶δ。这个依赖于pcna的系统修复了天然的AP位点和四氢fiiran残基。在不需要PCNA的反应中，DNA聚合酶β能够代替DNA聚合酶δ修复天然AP位点。DNA聚合酶α不支持两种AP位点的修复。这一结果表明，AP位点可以通过两种不同的途径进行修复，即pcna依赖途径和DNA聚合酶β依赖途径。
DNA damage frequently leads to the production of apurinic/apyrimidinic (AP) sites, which are presumed to be repaired through the base excision pathway. For detailed analyses of this repair mechanism, a synthetic analog of an AP site, 3-hydroxy-2-hydroxymethyltetrahydrofuran (tetrahydrofuran), has been employed in a model system. Tetrahydrofuran residues are efficiently repaired in aXenopus laevisoocyte extract in which most repair events involve ATP-dependent incorporation of no more than four nucleotides (Y. Matsumoto and D. F. Bogenhagen, Mol. Cell. Biol. 9:3750–3757, 1989; Y. Matsumoto and D. F. Bogenhagen, Mol. Cell. Biol. 11:4441–4447, 1991). Using a series of column chromatography procedures to fractionateX. laevisovarian extracts, we developed a reconstituted system of tetrahydrofuran repair with five fractions, three of which were purified to near homogeneity: proliferating cell nuclear antigen (PCNA), AP endonuclease, and DNA polymerase δ. This PCNA-dependent system repaired natural AP sites as well as tetrahydrofiiran residues. DNA polymerase β was able to replace DNA polymerase δ only for repair of natural AP sites in a reaction that did not require PCNA. DNA polymerase α did not support repair of either type of AP site. This result indicates that AP sites can be repaired by two distinct pathways, the PCNA-dependent pathway and the DNA polymerase β-dependent pathway.