Functional Analysis of Histone ADP-Ribosylation In Vitro and in Cells

Dan Huang, Andrea D. Edwards, Xuan Gong, W. Lee Kraus

Research output: Chapter in Book/Report/Conference proceedingChapter


Gene regulation in the nucleus requires precise control of the molecular processes that dictate how, when, and which genes are transcribed. The posttranslational modification (PTM) of histones in chromatin is an effective means to link cellular signaling to gene expression outcomes. The repertoire of histone PTMs includes phosphorylation, acetylation, methylation, ubiquitylation, and ADP-ribosylation (ADPRylation). ADPRylation is a reversible PTM that results in the covalent transfer of ADP-ribose units derived from NAD+ to substrate proteins on glutamate, aspartate, serine, and other amino acids. Histones were the first substrate proteins identified for ADPRylation, over five decades ago. Since that time, histone ADPRylation has been shown to be a widespread and critical regulator of chromatin structure and function during transcription, DNA repair, and replication. Here, we describe a set of protocols that allow the user to investigate site-specific histone ADPRylation and its functional consequences in biochemical assays and in cells in a variety of biological systems. With the recent discovery that some cancer-causing histone mutations (i.e., oncohistone mutations) occur at functional sites of regulatory ADPRylation, these protocols may have additional utility in studies of oncology.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages36
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • ADP-ribose (ADPR)
  • ADP-ribosylation (ADPRylation)
  • Chromatin
  • DNA repair
  • Histone
  • Oncohistone
  • Poly(ADP-ribosyl)ation (PARylation)
  • Posttranslational modification (PTM)
  • Transcription

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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