Abstract
Poly(ADP-ribose) Polymerase 1 (PARP-1) is an abundant chromatin associated protein, typical for most eukaryotic nuclei. The localization of PARP-1 in chromatin and its enzymatic activation involves multiple interactions of PARP-1 with nucleosomal histones, other proteins, and DNA. We report a set of methods designed to reconstitute PARP-1 regulation in vitro. These methods involve the expression of PARP-1 and PARP-1-regulating proteins using bacterial and eukaryotic systems, purification of these proteins using chromatography, testing of individual interactions in vitro, assembly of active complexes, and reconstitution of PARP-1 regulating reactions in vitro.
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Acknowledgements
Research was supported by grant from the National Science Foundation MCB-1616740 to A.V.T.
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Thomas, C., Kotova, E., Tulin, A.V. (2017). PARP-1 Interaction with and Activation by Histones and Nucleosomes. In: Tulin, A. (eds) Poly(ADP-Ribose) Polymerase. Methods in Molecular Biology, vol 1608. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6993-7_17
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DOI: https://doi.org/10.1007/978-1-4939-6993-7_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6992-0
Online ISBN: 978-1-4939-6993-7
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