HPF1 and nucleosomes mediate a dramatic switch in activity of PARP1 from polymerase to hydrolase

  1. Johannes Rudolph
  2. Genevieve Roberts
  3. Uma M Muthurajan
  4. Karolin Luger  Is a corresponding author
  1. University of Colorado, Boulder, United States
  2. HHMI and University of Colorado, Boulder, United States

Abstract

Poly(ADP-ribose) polymerase 1 (PARP1) is an important player in the response to DNA damage. Recently, histone PARylation factor (HPF1) was shown to be a critical modulator of the activity of PARP1 by facilitating PARylation of histones and redirecting the target amino acid specificity from acidic to serine residues. Here we investigate the mechanism and specific consequences of HPF1-mediated PARylation using nucleosomes as both activators and substrates for PARP1. HPF1 provides that catalytic base Glu284 to substantially redirect PARylation by PARP1 such that the histones in nucleosomes become the primary recipients of PAR chains. Surprisingly, HPF1 partitions most of the reaction product to free ADPR, resulting in much shorter PAR chains compared to reactions in the absence of HPF1. This HPF1-mediated switch from polymerase to hydrolase has important implications for the PARP1-mediated response to DNA damage and raises interesting new questions about the role of intracellular ADPR and depletion of NAD+.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for all Figures.

Article and author information

Author details

  1. Johannes Rudolph

    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0230-3323
  2. Genevieve Roberts

    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Uma M Muthurajan

    Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Karolin Luger

    Department of Chemistry and Biochemistry, HHMI and University of Colorado, Boulder, Boulder, United States
    For correspondence
    karolin.luger@colorado.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5136-5331

Funding

National Cancer Institute (CA218255)

  • Karolin Luger

Howard Hughes Medical Institute

  • Karolin Luger

National Institutes of Health (T32GM008759)

  • Genevieve Roberts

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Maria Spies, University of Iowa, United States

Version history

  1. Received: December 15, 2020
  2. Accepted: March 7, 2021
  3. Accepted Manuscript published: March 8, 2021 (version 1)
  4. Version of Record published: March 31, 2021 (version 2)

Copyright

© 2021, Rudolph et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Johannes Rudolph
  2. Genevieve Roberts
  3. Uma M Muthurajan
  4. Karolin Luger
(2021)
HPF1 and nucleosomes mediate a dramatic switch in activity of PARP1 from polymerase to hydrolase
eLife 10:e65773.
https://doi.org/10.7554/eLife.65773

Share this article

https://doi.org/10.7554/eLife.65773

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