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Biochemical and Biophysical Methods for Analysis of Poly(ADP-Ribose) Polymerase 1 and Its Interactions with Chromatin

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Poly(ADP-Ribose) Polymerase

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

Abstract

Poly (ADP-Ribose) Polymerase I (PARP-1) is a first responder to DNA damage and participates in the regulation of gene expression. The interaction of PARP-1 with chromatin and DNA is complex and involves at least two different modes of interaction. In its enzymatically inactive state, PARP-1 binds native chromatin with similar affinity as it binds free DNA ends. Automodification of PARP-1 affects interaction with chromatin and DNA to different extents. Here we describe a series of biochemical and biophysical techniques to quantify and dissect the different binding modes of PARP-1 with its various substrates. The techniques listed here allow for high throughput and quantitative measurements of the interaction of different PARP-1 constructs (inactive and automodified) with chromatin and DNA damage models.

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Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Colorado at Boulder, 3415 Colorado Avenue, Boulder, CO, 80303, USA

    Maggie H. Chassé, Uma M. Muthurajan & Karolin Luger

  2. Van Andel Research Institute, Grand Rapids, MI, USA

    Maggie H. Chassé

  3. Amgen Inc., Thousand Oaks, CA, USA

    Nicholas J. Clark

  4. Children’s Mercy Hospital, Kansas City, MO, USA

    Michael A. Kramer

  5. BioCAT, CSRRI and Department BCS, Illinois Institute of Technology, Chicago, IL, USA

    Srinivas Chakravarthy & Thomas Irving

  6. Howard Hughes Medical Institute, Chevy Chase, MD, USA

    Karolin Luger

  7. Institute for Genome Architecture and Function, Colorado State University, Fort Collins, CO, USA

    Karolin Luger

Authors
  1. Maggie H. Chassé
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  2. Uma M. Muthurajan
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  3. Nicholas J. Clark
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  4. Michael A. Kramer
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  5. Srinivas Chakravarthy
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  6. Thomas Irving
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  7. Karolin Luger
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Corresponding author

Correspondence to Karolin Luger .

Editor information

Editors and Affiliations

  1. Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA

    Alexei V. Tulin

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Chassé, M.H. et al. (2017). Biochemical and Biophysical Methods for Analysis of Poly(ADP-Ribose) Polymerase 1 and Its Interactions with Chromatin. 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_16

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  • DOI: https://doi.org/10.1007/978-1-4939-6993-7_16

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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

  • eBook Packages: Springer Protocols

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