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PARP-1 Interaction with and Activation by Histones and Nucleosomes

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1608)

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.

Key words

PARP-1 Nucleosomal histones Nucleosome Jil-1 kinase PARP-1 binding PARP-1 activation Poly(ADP-ribose) 

Notes

Acknowledgements

Research was supported by grant from the National Science Foundation MCB-1616740 to A.V.T.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Cancer Biology Program, Fox Chase Cancer CenterPhiladelphiaUSA

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