A method for evaluating nucleosome stability with a protein-binding fluorescent dye

Hiroyuki Taguchi; Naoki Horikoshi; Yasuhiro Arimura; Hitoshi Kurumizaka

doi:10.1016/j.ymeth.2014.08.019

A method for evaluating nucleosome stability with a protein-binding fluorescent dye

Methods. 2014 Dec;70(2-3):119-26. doi: 10.1016/j.ymeth.2014.08.019. Epub 2014 Sep 8.

Authors

Hiroyuki Taguchi¹, Naoki Horikoshi², Yasuhiro Arimura¹, Hitoshi Kurumizaka³

Affiliations

¹ Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan.
² Research Institute for Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan.
³ Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan; Research Institute for Science and Engineering, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480, Japan. Electronic address: kurumizaka@waseda.jp.

PMID: 25220913
DOI: 10.1016/j.ymeth.2014.08.019

Abstract

Nucleosomes are extremely stable histone-DNA complexes that form the building blocks of chromatin, which accommodates genomic DNA within the nucleus. The dynamic properties of chromatin play essential roles in regulating genomic DNA functions, such as DNA replication, recombination, repair, and transcription. Histones are the protein components of nucleosomes, and their diverse modifications and variants increase the versatility of nucleosome structures and their dynamics in chromatin. Therefore, a technique to evaluate the physical properties of nucleosomes would facilitate functional studies of the various nucleosomes. In this report, we describe a convenient assay for evaluating the thermal stability of nucleosomes in vitro.

Keywords: Chromatin; Histone octamer; Histone variant; Nucleosome; SYPRO Orange; Tetrasome.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Histones / chemistry*
Histones / metabolism
Microscopy, Fluorescence / methods*
Models, Chemical
Models, Molecular
Nucleosomes / chemistry*
Nucleosomes / metabolism
Temperature

Substances

Histones
Nucleosomes