TY - CHAP
T1 - Functional Analysis of Histone ADP-Ribosylation In Vitro and in Cells
AU - Huang, Dan
AU - Edwards, Andrea D.
AU - Gong, Xuan
AU - Kraus, W. Lee
N1 - Funding Information:
ADPRylation-related research in the Kraus lab is supported by grants from the NIH/NIDDK (R01 DK069710), NIH/NCI (R01 CA229487, R01 CA251943), US DOD Ovarian Cancer Research Program (OCRP) (W81XWH-20-OCRP-IIRA), and Cancer Prevention and Research Institute of Texas (CPRIT) (RP190236, RP220325), as well funds from the Cecil H. and Ida Green Center for Reproductive Biology Sciences Endowment to W.L.K.
Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - ADP-ribose (ADPR)
KW - ADP-ribosylation (ADPRylation)
KW - Chromatin
KW - DNA repair
KW - Histone
KW - Oncohistone
KW - Poly(ADP-ribosyl)ation (PARylation)
KW - Posttranslational modification (PTM)
KW - Transcription
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U2 - 10.1007/978-1-0716-2891-1_11
DO - 10.1007/978-1-0716-2891-1_11
M3 - Chapter
C2 - 36515836
AN - SCOPUS:85144586943
T3 - Methods in Molecular Biology
SP - 157
EP - 192
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -