Nucleotide excision repair in Human cell lines lacking both XPC and CSB proteins.
Nucleotide excision repair in Human cell lines lacking both XPC and CSB proteins.
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DOI：
10.1093/nar/gkad334
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发表时间：
2023-07-07
影响因子：
14.9
通讯作者：
Sancar, Aziz
中科院分区：
文献类型：
--
作者：
Lindsey-Boltz, Laura A.;Yang, Yanyan;Kose, Cansu;Deger, Nazli;Eynullazada, Khagani;Kawara, Hiroaki;Sancar, Aziz
核苷酸切除修复通过两种不同的子途径（全局修复和转录偶联修复 (TCR)）消除紫外线诱导的 DNA 损伤。大量研究表明，在人类和其他哺乳动物细胞系中，XPC 蛋白是通过全局修复修复非转录 DNA 造成的 DNA 损伤所必需的，而 CSB 蛋白是通过 TCR 修复转录 DNA 造成的损伤所必需的。因此，通常认为用 XPC−/−/CSB−/− 双突变体废除两条子通路将消除所有核苷酸切除修复。在这里，我们描述了三种不同的 XPC−/−/CSB−/− 人类细胞系的构建，与预期相反，它们执行 TCR。 XPC 和 CSB 基因在源自色素性干皮病患者以及正常人成纤维细胞的细胞系中发生突变，并使用非常灵敏的 XR-seq 方法在全基因组水平上分析修复情况。正如预测的那样，XPC−/− 细胞仅表现出 TCR，而 CSB−/− 细胞仅表现出全局修复。然而，XPC−/−/CSB−/− 双突变细胞系虽然修复大大减少，但仍表现出 TCR。突变 CSA 基因以产生三重突变体 XPC−/−/CSB−/−/CSA−/− 细胞系，消除了所有残留的 TCR 活性。总之，这些发现为哺乳动物核苷酸切除修复的机制特征提供了新的见解。敏感的全基因组修复图谱揭示了 XPC-/-/CSB-/- 细胞中意想不到的修复活性，这些细胞是为了消除核苷酸切除修复的两条子途径而构建的。
Nucleotide excision repair removes UV-induced DNA damage through two distinct sub-pathways, global repair and transcription-coupled repair (TCR). Numerous studies have shown that in human and other mammalian cell lines that the XPC protein is required for repair of DNA damage from nontranscribed DNA via global repair and the CSB protein is required for repair of lesions from transcribed DNA via TCR. Therefore, it is generally assumed that abrogating both sub-pathways with an XPC−/−/CSB−/− double mutant would eliminate all nucleotide excision repair. Here we describe the construction of three different XPC−/−/CSB−/− human cell lines that, contrary to expectations, perform TCR. The XPC and CSB genes were mutated in cell lines derived from Xeroderma Pigmentosum patients as well as from normal human fibroblasts and repair was analyzed at the whole genome level using the very sensitive XR-seq method. As predicted, XPC−/− cells exhibited only TCR and CSB−/− cells exhibited only global repair. However, the XPC−/−/CSB−/− double mutant cell lines, although having greatly reduced repair, exhibited TCR. Mutating the CSA gene to generate a triple mutant XPC−/−/CSB−/−/CSA−/− cell line eliminated all residual TCR activity. Together, these findings provide new insights into the mechanistic features of mammalian nucleotide excision repair. Sensitive whole-genome repair mapping reveals unexpected repair activity in XPC-/-/CSB-/- cells constructed to eliminate both sub pathways of nucleotide excision repair.
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DOI：
10.1016/0921-8777(91)90032-k
发表时间：
1991-11-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
BARRETT, SF;ROBBINS, JH;KRAEMER, KH
通讯作者：
KRAEMER, KH
影响因子：
4.8
作者：
Mu, D;Sancar, A
通讯作者：
Sancar, A
影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
16.6
作者：
Sancar, Aziz
通讯作者：
Sancar, Aziz
影响因子：
4.8
作者：
Mu, D;Hsu, DS;Sancar, A
通讯作者：
Sancar, A