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PARP10 Multi-Site Auto- and Histone MARylation Visualized by Acid-Urea Gel Electrophoresis

García-Saura, Antonio Ginés and Schüler, Herwig LU orcid (2021) In Cells 10(3).
Abstract

Poly-ADP-ribose polymerase (PARP)-family ADP-ribosyltransferases function in various signaling pathways, predominantly in the nucleus and cytosol. Although PARP inhibitors are in clinical practice for cancer therapy, the enzymatic activities of individual PARP family members are yet insufficiently understood. We studied PARP10, a mono-ADP-ribosyltransferase and potential drug target. Using acid-urea gel electrophoresis, we found that the isolated catalytic domain of PARP10 auto-ADP-ribosylates (MARylates) at eight or more acceptor residues. We isolated individual species with either singular or several modifications and then analyzed them by mass spectrometry. The results confirmed multi-site MARylation in a random order and identified... (More)

Poly-ADP-ribose polymerase (PARP)-family ADP-ribosyltransferases function in various signaling pathways, predominantly in the nucleus and cytosol. Although PARP inhibitors are in clinical practice for cancer therapy, the enzymatic activities of individual PARP family members are yet insufficiently understood. We studied PARP10, a mono-ADP-ribosyltransferase and potential drug target. Using acid-urea gel electrophoresis, we found that the isolated catalytic domain of PARP10 auto-ADP-ribosylates (MARylates) at eight or more acceptor residues. We isolated individual species with either singular or several modifications and then analyzed them by mass spectrometry. The results confirmed multi-site MARylation in a random order and identified four acceptor residues. The mutagenesis of singular acceptor residues had a minor impact on the overall auto-MARylation level and no effect on the MARylation of histone H3.1. Together, our results suggest that PARP10 automodification may have functions in the regulation of intramolecular or partner binding events, rather than of its enzymatic catalysis. This contributes to a better understanding of PARP10 functions, and, in the long run, to gauging the consequences of PARP inhibitor actions.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
acid-urea PAGE, ADP-ribosylation, ARTD10, histone H3, MARylation, mass spectrometry, PARP10
in
Cells
volume
10
issue
3
publisher
MDPI AG
external identifiers
  • pmid:33804157
  • scopus:85103863349
ISSN
2073-4409
DOI
10.3390/cells10030654
language
English
LU publication?
yes
id
fb82deef-8c28-4afa-ad85-d96e29e6719a
date added to LUP
2021-04-19 09:42:11
date last changed
2024-09-07 18:04:59
@article{fb82deef-8c28-4afa-ad85-d96e29e6719a,
  abstract     = {{<p>Poly-ADP-ribose polymerase (PARP)-family ADP-ribosyltransferases function in various signaling pathways, predominantly in the nucleus and cytosol. Although PARP inhibitors are in clinical practice for cancer therapy, the enzymatic activities of individual PARP family members are yet insufficiently understood. We studied PARP10, a mono-ADP-ribosyltransferase and potential drug target. Using acid-urea gel electrophoresis, we found that the isolated catalytic domain of PARP10 auto-ADP-ribosylates (MARylates) at eight or more acceptor residues. We isolated individual species with either singular or several modifications and then analyzed them by mass spectrometry. The results confirmed multi-site MARylation in a random order and identified four acceptor residues. The mutagenesis of singular acceptor residues had a minor impact on the overall auto-MARylation level and no effect on the MARylation of histone H3.1. Together, our results suggest that PARP10 automodification may have functions in the regulation of intramolecular or partner binding events, rather than of its enzymatic catalysis. This contributes to a better understanding of PARP10 functions, and, in the long run, to gauging the consequences of PARP inhibitor actions.</p>}},
  author       = {{García-Saura, Antonio Ginés and Schüler, Herwig}},
  issn         = {{2073-4409}},
  keywords     = {{acid-urea PAGE; ADP-ribosylation; ARTD10; histone H3; MARylation; mass spectrometry; PARP10}},
  language     = {{eng}},
  number       = {{3}},
  publisher    = {{MDPI AG}},
  series       = {{Cells}},
  title        = {{PARP10 Multi-Site Auto- and Histone MARylation Visualized by Acid-Urea Gel Electrophoresis}},
  url          = {{http://dx.doi.org/10.3390/cells10030654}},
  doi          = {{10.3390/cells10030654}},
  volume       = {{10}},
  year         = {{2021}},
}