Craig Ashley L, Moser Sandra C, Bailly Aymeric P, Gartner Anton
Wellcome Trust Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Scotland, United Kingdom.
Methods Cell Biol. 2012;107:321-52. doi: 10.1016/B978-0-12-394620-1.00011-4.
In response to genotoxic insults, cells activate DNA damage response pathways that either stimulate transient cell cycle arrest and DNA repair or induce apoptosis. The Caenorhabditis elegans germ line is now well established as a model system to study these processes in a genetically tractable, multicellular organism. Upon treatment with genotoxic agents, premeiotic C. elegans germ cells transiently halt cell cycle progression, whereas meiotic prophase germ cells in the late-pachytene stage undergo apoptosis. Further, accumulation of unrepaired meiotic recombination intermediates can also lead to apoptosis of affected pachytene cells. DNA damage-induced cell death requires key components of the evolutionarily conserved apoptotic machinery. Moreover, both cell cycle arrest and pachytene apoptosis responses depend on conserved DNA damage checkpoint proteins. Genetics- and genomics-based approaches that have demonstrated roles for conserved checkpoint proteins have also begun to uncover novel components of these response pathways. In this chapter, we briefly review the C. elegans DNA damage response field, discuss in detail methods currently used to assay DNA damage responses in C. elegans, and describe the development of new experimental tools that will facilitate a more comprehensive understanding of the DNA damage response.
作为对基因毒性损伤的反应，细胞会激活DNA损伤反应通路，该通路要么刺激短暂的细胞周期停滞和DNA修复，要么诱导细胞凋亡。秀丽隐杆线虫的生殖系现已成为一个成熟的模型系统，用于在遗传上易于处理的多细胞生物体中研究这些过程。在用基因毒性试剂处理后，减数分裂前的秀丽隐杆线虫生殖细胞会短暂地停止细胞周期进程，而处于粗线期后期的减数分裂前期生殖细胞则会发生凋亡。此外，未修复的减数分裂重组中间体的积累也会导致受影响的粗线期细胞凋亡。DNA损伤诱导的细胞死亡需要进化上保守的凋亡机制的关键成分。此外，细胞周期停滞和粗线期凋亡反应都依赖于保守的DNA损伤检查点蛋白。基于遗传学和基因组学的方法已证明保守检查点蛋白的作用，这些方法也已开始揭示这些反应通路的新成分。在本章中，我们简要回顾秀丽隐杆线虫DNA损伤反应领域，详细讨论目前用于检测秀丽隐杆线虫DNA损伤反应的方法，并描述新实验工具的开发，这些工具将有助于更全面地理解DNA损伤反应。
