Using single-molecule approach to visualize the nucleosome assembly in yeast nucleoplasmic extracts
Using single-molecule approach to visualize the nucleosome assembly in yeast nucleoplasmic extracts
复制标题
使用单分子方法可视化酵母核质提取物中的核小体组装
DOI：
10.1016/j.scib.2017.02.011
复制
发表时间：
2017-01-01
期刊：
影响因子：
18.9
通讯作者：
Fu, Yu V.
中科院分区：
文献类型：
--
作者：
Chen, Xiuqiang;Zhao, Ershuang;Fu, Yu V.
在真核细胞中，染色质的最小亚基是核小体，它由缠绕在组蛋白核心上的DNA片段组成。尽管经过多年的努力，核小体的组装和拆卸过程仍然不是很清楚。在这里，我们提出了一个方便的方法来研究核小体组装过程中的单分子水平。我们发明了一种新的酵母核质提取物系统（YNPE），并证明了YNPE在生理条件下支持核小体的组装。将全内反射荧光显微镜技术与微流控流动池技术相结合，从单分子水平观察了核小体在YNPE中组装的动态过程。我们的系统提供了一种新的体外单分子工具，研究在生理条件下的核小体组装的动力学。(C)中国科学出版社.爱思唯尔公司、科学中国出版社出版。All rights reserved.
In eukaryotic cells, the smallest subunit of chromatin is the nucleosome, which consists of a segment of DNA wound on histone protein cores. Despite many years of effort, the process of nucleosome assembly and disassembly is still not very clear. Here, we present a convenient method to investigate the process of nucleosome assembly at the single molecule level. We invented a novel system derived from the yeast nucleoplasmic extracts (YNPE), and demonstrated that the YNPE supports the nucleosome assembly under physiological condition. By combining the total internal reflection fluorescence microscopy with microfluidic flow-cell technique, the dynamic process of nucleosome assembly in YNPE was visualized at single-molecule level. Our system provides a novel in vitro single-molecule tool to investigate the dynamics of nucleosome assembly under physiological conditions. (C) 2017 Science China Press. Published by Elsevier B.V. and Science China Press. All rights reserved.