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Modular antibodies reveal DNA damage-induced mono-ADP-ribosylation as a second wave of PARP1 signaling

Edoardo José Longarini1, Helen Dauben1, Carolina Locatelli1

  • 1Research Group of Proteomics and ADP-Ribosylation Signaling, Max Planck Institute for Biology of Ageing, 50931 Cologne, Germany.

Molecular Cell
|April 28, 2023
PubMed

Insights

Mono-ADP-ribosylation, a signaling pathway involving HPF1/PARP1, actively participates in cellular responses to DNA damage. This study developed new tools to detect this signal, revealing its role in DNA repair and telomere maintenance.

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Poly(ADP-ribosyl)transferase (PARP1) is a key regulator of DNA repair and an anti-cancer target.
  • Mono-ADP-ribosylation's role in DNA damage response was previously unclear, with its active function unknown.

Purpose of the Study:

  • To investigate the functional role of mono-ADP-ribosylation in cellular signaling, particularly after DNA damage.
  • To develop novel detection methods for mono-ADP-ribosylation, enabling live-cell imaging and proteomic analysis.

Main Methods:

  • Engineering SpyTag-based modular antibodies for sensitive mono-ADP-ribosylation detection.
  • Developing fluorescence-based sensors for live-cell imaging of mono-ADP-ribosylation.
  • Utilizing multilevel chromatin proteomics to identify mono-ADP-ribosylation readers.

Main Results:

  • Serine mono-ADP-ribosylation represents a distinct signaling wave mediated by the HPF1/PARP1 ratio.
  • Histone mono-ADP-ribosylation readers, such as RNF114, were identified and shown to be recruited to DNA lesions.
  • RNF114's role in modulating DNA damage response and telomere maintenance was demonstrated.

Conclusions:

  • Mono-ADP-ribosylation by HPF1/PARP1 is an active and important cellular signal, not merely a byproduct.
  • The developed technological framework facilitates broader research into ADP-ribosylation.
  • This work establishes a new understanding of DNA damage response pathways involving mono-ADP-ribosylation.
Keywords:
ADP-ribosylationDNA damage responseHPF1PARP1RNF114SpyTagantibodiestelomere

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