In vitro ubiquitination and degradation of bacteria-purified human histone H2B by the nucleoli fractions

Sheng Li; Guoan Zhang; Yanping Liu; Qinru Sun; Lu Yang; Junli Dong; Bowen Zhang; Su Chen

doi:10.1016/j.xpro.2021.100601

In vitro ubiquitination and degradation of bacteria-purified human histone H2B by the nucleoli fractions

STAR Protoc. 2021 Jun 11;2(2):100601. doi: 10.1016/j.xpro.2021.100601. eCollection 2021 Jun 18.

Authors

Sheng Li¹, Guoan Zhang¹, Yanping Liu², Qinru Sun², Lu Yang³, Junli Dong³, Bowen Zhang³, Su Chen³

Affiliations

¹ School of Forensic Sciences and Laboratory Medicine, Jining Medical University, Jining, Shandong 272067, PR China.
² School of Forensic Sciences, Xi'an Jiao Tong University Health Science Center, Xi'an, Shaanxi 710061, PR China.
³ Laboratory of Molecular and Cellular Biology, School of Basic Medical Sciences, Henan University School of Medicine, Kaifeng, Henan 475004, PR China.

Abstract

Histones are the main components of chromatin, and the protein levels of histones significantly affect chromatin assembly. Here, we describe detailed protocols for histone H2B purification from bacteria and for the separation of nucleolar fractions and cytoplasmic and nucleoplasmic fractions. Finally, the in vitro ubiquitination and degradation of H2B by distinct cellular fractions are described. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).

Keywords: Cancer; Cell separation/fractionation; Molecular Biology; Protein expression and purification.

Publication types

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

MeSH terms

Bacteria / metabolism*
Cell Nucleolus / metabolism*
Histones / metabolism*
Humans
Hydrolysis
In Vitro Techniques
Ubiquitination

Substances

Histones