Abstract
The catalytic activity of histone methyltransferases is not restricted to histones but also includes noncanonical substrates. Increasing evidence shows that histone methyltransferases methylate themselves, and automethylation has emerged as a self-regulatory mechanism. Here, we introduce experimental procedures to identify automethylation sites of histone methyltransferases and to investigate the function of automethylation in a reconstituted biochemical system and in cellular contexts.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Lee C-H, Yu J-R, Granat J et al (2019) Automethylation of PRC2 promotes H3K27 methylation and is impaired in H3K27M pediatric glioma. Genes Dev 33:1428–1440
Wang X, Long Y, Paucek RD et al (2019) Regulation of histone methylation by automethylation of PRC2. Genes Dev 33:1416–1427
Piao L, Nakakido M, Suzuki T, Dohmae N, Nakamura Y, Hamamoto R (2016) Automethylation of SUV39H2, an oncogenic histone lysine methyltransferase, regulates its binding affinity to substrate proteins. Oncotarget 7:22846–22856
Iglesias N, Currie MA, Jih G et al (2018) Automethylation-induced conformational switch in Clr4 (Suv39h) maintains epigenetic stability. Nature 560:504–508
Chin HG, Estève P-O, Pradhan M et al (2007) Automethylation of G9a and its implication in wider substrate specificity and HP1 binding. Nucleic Acids Res 35:7313–7323
Poulard C, Baulu E, Lee BH, Pufall MA, Stallcup MR (2018) Increasing G9a automethylation sensitizes B acute lymphoblastic leukemia cells to glucocorticoid-induced death. Cell Death Dis 9:1038
Nishioka K, Reinberg D (2003) Methods and tips for the purification of human histone methyltransferases. Methods 31:49–58
Yun M, Ruan C, Huh JW, Li B (2012) Reconstitution of modified chromatin templates for in vitro functional assays. Methods Mol Biol 833:237–253
Kurien BT, Scofield RH (2015) Multiple immunoblots by passive diffusion of proteins from a single SDS-PAGE gel. Methods Mol Biol 1312:77–86
Li F, Mullins JI (2002) Site-directed mutagenesis facilitated by DpnI selection on hemimethylated DNA. Methods Mol Biol 182:19–27
Acknowledgments
We thank Dr. D. Reinberg for critical reading of the manuscript. This work was supported by Research Resettlement Fund for the new faculty of Seoul National University, Creative-Pioneering Researchers Program through Seoul National University, grants from Seoul National University College of Medicine, National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2021R1C1C1013220), and BK21 Four Biomedical Science Program. L. Popoca is supported by National Institutes of Health (NIH) grant R01CA199652, the Howard Hughes Medical Institute (HHMI), and the Making Headway Foundation St. Baldrick’s Research Grant (189290).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Popoca, L., Lee, CH. (2022). Techniques to Study Automethylation of Histone Methyltransferases and its Functional Impact. In: Margueron, R., Holoch, D. (eds) Histone Methyltransferases. Methods in Molecular Biology, vol 2529. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2481-4_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2481-4_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2480-7
Online ISBN: 978-1-0716-2481-4
eBook Packages: Springer Protocols