检测与双链断裂的基因转换修复相关的突变

Assaying Mutations Associated With Gene Conversion Repair of a Double-Strand Break.

作者信息

Dwivedi Gajendrahar, Haber James E

机构信息

Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA, United States.

Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA, United States.

出版信息

Methods Enzymol. 2018;601:145-160. doi: 10.1016/bs.mie.2017.11.029. Epub 2018 Feb 28.

Abstract

DNA double-strand break (DSB) is a cytotoxic lesion and needs to be repaired immediately. There are several metabolic pathways evolved to repair a DSB. Gene conversion is one of the least error-prone pathway for repair of a DNA DSB. Despite this there is nearly 1000-fold increase in mutation rate associated with gene conversion. Not only higher mutation rate is associated with gene conversion but also there is a very distinct mutation profile compared to spontaneous mutation events. Gene conversion is characterized by the presence of very high frameshift mutation events and other complex mutations that are not present during regular DNA replication. Another DNA DSB repair pathway widely studied is "break-induced replication" (BIR). BIR has been shown to be highly mutagenic in nature. BIR may lead to chromosomal rearrangement and has potential to cause cluster mutations with serious disease implications. In this chapter, the design of assay systems to study various mutation types and experimental procedures to measure specific mutation frequency associated with gene conversion are discussed.

摘要

DNA双链断裂(DSB)是一种细胞毒性损伤,需要立即修复。进化出了几种代谢途径来修复DSB。基因转换是修复DNA DSB最不易出错的途径之一。尽管如此,与基因转换相关的突变率仍增加了近1000倍。不仅基因转换与更高的突变率相关,而且与自发突变事件相比,还有非常独特的突变谱。基因转换的特征是存在非常高的移码突变事件和其他在常规DNA复制过程中不存在的复杂突变。另一种被广泛研究的DNA DSB修复途径是“断裂诱导复制”(BIR)。BIR已被证明在本质上具有高度致突变性。BIR可能导致染色体重排,并有可能引起具有严重疾病影响的簇状突变。在本章中,将讨论用于研究各种突变类型的检测系统设计以及测量与基因转换相关的特定突变频率的实验程序。

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