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

STAR Protocols (Jun 2021)

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

Affiliations

Sheng Li: School of Forensic Sciences and Laboratory Medicine, Jining Medical University, Jining, Shandong 272067, PR China
Guoan Zhang: School of Forensic Sciences and Laboratory Medicine, Jining Medical University, Jining, Shandong 272067, PR China
Yanping Liu: School of Forensic Sciences, Xi’an Jiao Tong University Health Science Center, Xi’an, Shaanxi 710061, PR China
Qinru Sun: School of Forensic Sciences, Xi’an Jiao Tong University Health Science Center, Xi’an, Shaanxi 710061, PR China
Lu Yang: Laboratory of Molecular and Cellular Biology, School of Basic Medical Sciences, Henan University School of Medicine, Kaifeng, Henan 475004, PR China
Junli Dong: Laboratory of Molecular and Cellular Biology, School of Basic Medical Sciences, Henan University School of Medicine, Kaifeng, Henan 475004, PR China
Bowen Zhang: Laboratory of Molecular and Cellular Biology, School of Basic Medical Sciences, Henan University School of Medicine, Kaifeng, Henan 475004, PR China
Su Chen: Laboratory of Molecular and Cellular Biology, School of Basic Medical Sciences, Henan University School of Medicine, Kaifeng, Henan 475004, PR China; Corresponding author

Journal volume & issue: Vol. 2, no. 2
p. 100601

Abstract

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Summary: 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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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