A combinatorial system to examine the enzymatic repair of multiply damaged DNA substrates.
A combinatorial system to examine the enzymatic repair of multiply damaged DNA substrates.
复制标题
用于检查多重损伤 DNA 底物酶促修复的组合系统。
DOI：
10.1093/nar/gkac530
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发表时间：
2022-07-22
影响因子：
14.9
通讯作者：
Sowers, Lawrence C.
中科院分区：
文献类型：
--
作者：
Hsu, Chia Wei;Conrad, James W.;Sowers, Mark L.;Baljinnyam, Tuvshintugs;Herring, Jason L.;Hackfeld, Linda C.;Hatch, Sandra S.;Sowers, Lawrence C.
DNA损伤驱动基因突变，这些基因突变是人类癌症发展的基础。在哺乳动物细胞中已经描述了多种可以修复这种损伤的途径。然而，迄今为止，大多数工作都集中在DNA中的单个病变上。我们在这里提出了一个组合系统，它允许通过连接在一起的六个寡核苷酸含有损坏或修饰的碱基含有单一或多种类型的损伤的双链体的组装。该组合系统具有双荧光标记，允许同时检查两条链，以研究不同DNA修复途径之间的相互作用或竞争。使用这个系统，我们展示了如何修复一个DNA链中的氧化损伤可以将错配的T：G脱氨基中间体转化为T：A突变。我们还表明，缓慢修复的T：G错配，相对于U：G错配，由人类甲基结合结构域4 DNA糖基化酶提供了一个竞争优势，竞争修复途径，并可以解释为什么CpG二核苷酸是热点的C到T突变在人类肿瘤。数据还表明，修复相对链的紧密间隔的病变可以修复的组合，短和长补丁的基础切除修复和多重损伤位点的同时修复可能会导致致命的双链断裂。
DNA damage drives genetic mutations that underlie the development of cancer in humans. Multiple pathways have been described in mammalian cells which can repair this damage. However, most work to date has focused upon single lesions in DNA. We present here a combinatorial system which allows assembly of duplexes containing single or multiple types of damage by ligating together six oligonucleotides containing damaged or modified bases. The combinatorial system has dual fluorescent labels allowing examination of both strands simultaneously, in order to study interactions or competition between different DNA repair pathways. Using this system, we demonstrate how repair of oxidative damage in one DNA strand can convert a mispaired T:G deamination intermediate into a T:A mutation. We also demonstrate that slow repair of a T:G mispair, relative to a U:G mispair, by the human methyl-binding domain 4 DNA glycosylase provides a competitive advantage to competing repair pathways, and could explain why CpG dinucleotides are hotspots for C to T mutations in human tumors. Data is also presented that suggests repair of closely spaced lesions in opposing strands can be repaired by a combination of short and long-patch base excision repair and simultaneous repair of multiply damage sites can potentially lead to lethal double strand breaks.
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影响因子：
14.9
作者：
Grin I;Ishchenko AA
通讯作者：
Ishchenko AA
DOI：
10.1016/j.jbc.2022.101638
发表时间：
2022-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hsu CW;Sowers ML;Baljinnyam T;Herring JL;Hackfeld LC;Tang H;Zhang K;Sowers LC
通讯作者：
Sowers LC
DOI：
10.1073/pnas.131077798
发表时间：
2001-06-19
影响因子：
11.1
作者：
Blaisdell, JO;Wallace, SS
通讯作者：
Wallace, SS
影响因子：
4.1
作者：
Gates, Kent S.
通讯作者：
Gates, Kent S.
影响因子：
7.4
作者：
Cadet J;Davies KJA;Medeiros MH;Di Mascio P;Wagner JR
通讯作者：
Wagner JR