In Vitro and In Vivo Assays for Studying Histone Ubiquitination and Deubiquitination

Joo, Heui-Yun; Wang, Hengbin; Zhai, Ling

doi:10.1007/978-1-59745-190-1_20

Cited by 2 publications

(1 citation statement)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this paper we have detailed the experimental procedures for biochemical purification of histone ubiquitin ligases from HeLa cells. Similar biochemical approaches have used to identify enzymes involved in histone methylation, demethylation, deubiquitination, and other processes (29–31). This approach is an unbiased, systemic search for novel histone modifying enzyme activities.…”

Section: Discussionmentioning

confidence: 99%

Purification of histone ubiquitin ligases from HeLa cells

Jones¹,

Joo²,

Robbins³

et al. 2011

Methods

Self Cite

View full text Add to dashboard Cite

Posttranslational histone modifications play an important role in regulating chromatin based nuclear processes including transcription. Of these modifications, histone ubiquitination is among the least understood. Histone ubiquitination predominately targets histones H2A and H2B. While ubiquitination of H2B is evolutionarily conserved from budding yeast to mammals, ubiquitination of H2A has not been detected in budding yeast, worms, or plants. Until recently, studies of histone ubiquitination lagged far behind the study of other histone modifications, largely because antibodies specific for ubiquitinated histones are difficult to generate. Despite this obstacle, the identification of the enzymatic machineries involved in histone ubiquitination, together with the successful use of a combination of genetic and immunoblot approaches to detect ubiquitinated histones, have helped to reveal important regulatory roles for this modification in transcriptional initiation and elongation, cell cycle progression, and DNA damage response. With the aid of the recently developed ubiquitinated histone-specific antibodies, an intriguing link between histone ubiquitination and cancer development has been established. While the enzymes involved in H2B ubiquitination were identified first in budding yeast and subsequently in higher organisms based on gene homology, the identification of the enzymatic machineries involved in H2A ubiquitination largely depended on a biochemical purification approach. The unbiased search for ubiquitin ligases targeting histones also led to the identification of a H3 and H4 ubiquitin ligase. Here we detail a protocol for the biochemical approach to identify histone ubiquitin ligase(s) from HeLa cells. Similar approaches have been successfully used to identify histone methyltransferases, histone demethylases, chromatin remodeling factors, and general transcription factors. So long as an in vitro enzymatic assay can be established, the approach we describe can be easily adapted to identify other histone and non-histone modifying enzymes.

show abstract

Section: Discussionmentioning

confidence: 99%

Purification of histone ubiquitin ligases from HeLa cells

Jones¹,

Joo²,

Robbins³

et al. 2011

Methods

Self Cite

View full text Add to dashboard Cite

show abstract

An Overview of HDAC Inhibitors and their Synthetic Routes

Peng

Liao

Sun

et al. 2019

CTMC

View full text Add to dashboard Cite

Epigenetics play a key role in the origin, development and metastasis of cancer. Epigenetic processes include DNA methylation, histone acetylation, histone methylation, and histone phosphorylation, among which, histone acetylation is the most common one that plays important roles in the regulation of normal cellular processes, and is controlled by histone deacetylases (HDACs) and histone acetyltransferases (HATs). HDACs are involved in the regulation of many key cellular processes, such as DNA damage repair, cell cycle control, autophagy, metabolism, senescence and chaperone function, and can lead to oncogene activation. As a result, HDACs are considered to be an excellent target for anti-cancer therapeutics like histone deacetylase inhibitors (HDACi) which have attracted much attention in the last decade. A wide-ranging knowledge of the role of HDACs in tumorigenesis, and of the action of HDACi, has been achieved. The primary purpose of this paper is to summarize recent HDAC inhibitors and the synthetic routes as well as to discuss the direction for the future development of new HDAC inhibitors.

show abstract

In Vitro and In Vivo Assays for Studying Histone Ubiquitination and Deubiquitination

Cited by 2 publications

References 35 publications

Purification of histone ubiquitin ligases from HeLa cells

Purification of histone ubiquitin ligases from HeLa cells

An Overview of HDAC Inhibitors and their Synthetic Routes

Contact Info

Product

Resources

About