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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References
Feng FY, de Bono JS, Rubin MA, Knudsen KE (2015) Chromatin to clinic: the molecular rationale for PARP1 inhibitor function. Mol Cell 58:925–934
Thomas C, Tulin A (2013) Poly-ADP-ribose polymerase: machinery for nuclear processes. Mol Asp Med 34:1124–1137
Pinnola A, Naumova N, Shah M, Tulin AV (2007) Nucleosomal core histones mediate dynamic regulation of poly(ADP-ribose) polymerase 1 protein binding to chromatin and induction of its enzymatic activity. J Biol Chem 282:32511–32519
Tulin A, Spradling A (2003) Chromatin loosening by poly(ADP)-ribose polymerase (PARP) at drosophila puff loci. Science 299:560–562
Kotova E et al (2011) Drosophila histone H2A variant (H2Av) controls poly(ADP-ribose) polymerase 1 (PARP1) activation in chromatin. Proc Natl Acad Sci 108:6205–6210
Thomas CJ et al (2014) Kinase-mediated changes in nucleosome conformation trigger chromatin decondensation via poly(ADP-ribosyl)ation. Mol Cell 53:831–842
Luger K, Rechsteiner TJ, Richmond TJ (1999) Expression and purification of recombinant histones and nucleosome reconstitution. Methods Mol Biol 119:1–16
Jin Y et al (1999) JIL-1: a novel chromosomal tandem kinase implicated in transcriptional regulation in Drosophila. Mol Cell 4:129–135
Cirillo LA et al (2002) Opening of compacted chromatin by early developmental transcription factors HNF3 (FoxA) and GATA-4. Mol Cell 9:279–289
Kirsanov KI et al (2014) Minor groove binding ligands disrupt PARP-1 activation pathways. Oncotarget 5:428–437
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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