Generation and Analysis of dsDNA Breaks for Checkpoint and Repair Studies in Fission Yeast.
Generation and Analysis of dsDNA Breaks for Checkpoint and Repair Studies in Fission Yeast.
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用于裂殖酵母检查点和修复研究的 dsDNA 断裂的产生和分析。
DOI：
10.1007/978-1-0716-1217-0_13
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
O'Connell,MatthewJ
中科院分区：
文献类型：
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作者：
Ramalingam,Rohana;O'Connell,MatthewJ
DNA的损伤会导致检查点和修复反应。这些都是涉及许多基因的复杂事件，这些基因的产物在病变处组装并形成信号级联以募集额外的因子并调节细胞周期。裂殖酵母已被证明是一个很好的模型来研究这些事件，并导致基因和途径发现的努力。最近的进展涉及对病变最早期事件的更详细分析，特别是双链DNA断裂（DSB）。在这里，我们描述了几种方法来分析事件在DSB，无论是对DNA和招募蛋白质这些病变，使用S。pombeas一个模型。然而，这些方法中的每一种都很容易适用于对协议进行微小修改的任何实验系统。
Damage to DNA elicits both checkpoint and repair responses. These are complex events that involve many genes whose products assemble at lesions and form signaling cascades to recruit additional factors and regulate the cell cycle. The fission yeastSchizosaccharomyces pombehas proven to be an excellent model to study these events, and has led gene and pathway discovery efforts. Recent progress has involved a more detailed analysis of the earliest events at lesions, particularly double-stranded DNA breaks (DSBs). Here we describe several methods for the analysis of events at DSBs, both on the DNA and the recruitment of proteins to these lesions, usingS. pombeas a model. However, each of these methods is easily applicable to any experimental system with minor modifications to the protocols.