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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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Correspondence to Michael O. Hottiger .

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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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