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.
CITATION STYLE
Thomas, C., Kotova, E., & Tulin, A. V. (2017). PARP-1 interaction with and activation by histones and nucleosomes. In Methods in Molecular Biology (Vol. 1608, pp. 255–267). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6993-7_17
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