Studying the organization of DNA repair by single-cell and single-molecule imaging.
Studying the organization of DNA repair by single-cell and single-molecule imaging.
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DOI：
10.1016/j.dnarep.2014.02.015
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发表时间：
2014-08
期刊：
影响因子：
3.8
通讯作者：
Kapanidis, Achillefs N.
中科院分区：
文献类型：
--
作者：
Uphoff, Stephan;Kapanidis, Achillefs N.
关键词：
单细胞实验研究DNA修复中的随机事件和异质性。定量DNA修复蛋白在细胞中的浓度、扩散和定位。使用光活化单分子追踪直接观察DNA修复。DNA修复保护基因组免受多种DNA损伤剂的侵害。虽然许多修复蛋白的机制已经在体外分别进行了研究，但对整个修复机制在体内的协调功能知之甚少。此外，单细胞研究表明，DNA损伤反应在细胞间的修复活动中产生了很大的变化。这篇评论的重点是荧光成像方法，通过测量蛋白质浓度，扩散特性，定位，相互作用和酶促速率提供了一个定量描述的DNA修复在单细胞中。新兴的单分子和超分辨率显微镜方法现在允许直接观察体内的单个蛋白质和DNA修复事件。我们希望通过将细胞异质性与分子水平的机制观察联系起来，可以了解更多关于DNA修复组织的信息。
Single-cell experiments to study stochastic events and heterogeneity in DNA repair. Quantifying DNA repair protein concentration, diffusion, and localization in cells. Direct observation of DNA repair using photoactivated single-molecule tracking. DNA repair safeguards the genome against a diversity of DNA damaging agents. Although the mechanisms of many repair proteins have been examined separately in vitro, far less is known about the coordinated function of the whole repair machinery in vivo. Furthermore, single-cell studies indicate that DNA damage responses generate substantial variation in repair activities across cells. This review focuses on fluorescence imaging methods that offer a quantitative description of DNA repair in single cells by measuring protein concentrations, diffusion characteristics, localizations, interactions, and enzymatic rates. Emerging single-molecule and super-resolution microscopy methods now permit direct visualization of individual proteins and DNA repair events in vivo. We expect much can be learned about the organization of DNA repair by linking cell heterogeneity to mechanistic observations at the molecular level.
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