Abstract
In vitro histone modification (HM) assays are used to characterize the activity of chromatin-modifying enzymes. These assays provide information regarding the modification sites on histones, the product specificity, and the impact of other histone or nucleotide modifications on enzyme activity. In particular, histone methyltransferase (HMT) assays have been instrumental in elucidating the activity and site specificity of many plant HMT enzymes. In this chapter, we describe a general protocol that can be used to perform HMT assays using different chromatin substrates, detection methods, and enzymes directly purified from plant material or heterologous sources.
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Acknowledgments
We want to thank Jean-François Couture (University of Ottawa) for his advice on some of the assays described and Chantal LeBlanc for her helpful comments on this chapter. Work in the Voigt lab is supported by the Wellcome Trust and the Royal Society (joint Grant Ref. 104175/Z/14/Z, Sir Henry Dale Fellowship to PV) and has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (ERC-STG grant agreement No. 639253). The Wellcome Trust Centre for Cell Biology is supported by core funding from the Wellcome Trust (Grant Ref. 092076).
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Jacob, Y., Voigt, P. (2018). In Vitro Assays to Measure Histone Methyltransferase Activity Using Different Chromatin Substrates. In: Bemer, M., Baroux, C. (eds) Plant Chromatin Dynamics. Methods in Molecular Biology, vol 1675. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7318-7_20
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DOI: https://doi.org/10.1007/978-1-4939-7318-7_20
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