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Subcellular Quantitation of ADP-Ribosylation by High-Content Microscopy

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Poly(ADP-Ribose) Polymerase

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2609))

Abstract

ADP-ribosylation is a posttranslational modification with many functions ranging from the DNA damage response to transcriptional regulation. While nuclear ADP-ribosylation has been extensively studied in the context of genotoxic stress mediated by PARP1, signaling by other members of the family and in other cellular compartments is still not as well understood. In recent years, however, progress has been made with the development of new tools for detection of ADP-ribosylation by immunofluorescence, which allows for a spatial differentiation of signal intensity for different cellular compartments. Here, we present our method for the detection and quantification of compartment-specific ADP-ribosylation by immunofluorescence and show why the engineered macrodomain eAf5121 might be the best tool to date.

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Acknowledgments

We thank Tobias Suter and Dr. Lorenza Ferretti (both from the Department of Molecular Mechanisms of Disease, University of Zurich) for the helpful discussion, thorough proofreading, and for editorial assistance. ADP-ribosylation research in the laboratory of M.O.H. is funded by the Kanton of Zurich and the Swiss National Science Foundation (grant no. 310030_205202 and IZLIZ3_200237).

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Author notes

  1. Authors Lukas Muskalla and Anka Güldenpfennig have contributed equally to this chapter.

Authors and Affiliations

  1. Department of Molecular Mechanisms of Disease, Vetsuisse Faculty and Faculty of Science, University of Zurich, Zurich, Switzerland

    Lukas Muskalla, Anka Güldenpfennig & Michael O. Hottiger

  2. Life Science Zurich Graduate School, Cancer Biology PhD program, University of Zurich, Zurich, Switzerland

    Lukas Muskalla

  3. Life Science Zurich Graduate School, Molecular Life Science PhD program, University of Zurich, Zurich, Switzerland

    Anka Güldenpfennig

Authors
  1. Lukas Muskalla
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  2. Anka Güldenpfennig
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  3. Michael O. Hottiger
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Corresponding author

Correspondence to Michael O. Hottiger .

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Editors and Affiliations

  1. Department of Biomedical Sciences, University of North Dakota, Grand Forks, ND, USA

    Alexei V. Tulin

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Muskalla, L., Güldenpfennig, A., Hottiger, M.O. (2023). Subcellular Quantitation of ADP-Ribosylation by High-Content Microscopy. In: Tulin, A.V. (eds) Poly(ADP-Ribose) Polymerase. Methods in Molecular Biology, vol 2609. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2891-1_7

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  • DOI: https://doi.org/10.1007/978-1-0716-2891-1_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2890-4

  • Online ISBN: 978-1-0716-2891-1

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