Using single-molecule approach to visualize the nucleosome assembly in yeast nucleoplasmic extracts

Xiuqiang Chen; Ershuang Zhao; Yu V Fu

doi:10.1016/j.scib.2017.02.011

Using single-molecule approach to visualize the nucleosome assembly in yeast nucleoplasmic extracts

Sci Bull (Beijing). 2017 Mar 30;62(6):399-404. doi: 10.1016/j.scib.2017.02.011. Epub 2017 Feb 24.

Authors

Xiuqiang Chen¹, Ershuang Zhao¹, Yu V Fu²

Affiliations

¹ State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China.
² State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: fuyu@im.ac.cn.

PMID: 36659283
DOI: 10.1016/j.scib.2017.02.011

Abstract

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.

Keywords: Microfluidic flow-cell; Nucleosome assembly; Single-molecule; Total internal reflection fluorescence microscopy; Yeast nucleoplasmic extracts.