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PARP14 is a PARP with both ADP-ribosyl transferase and hydrolase activities

Nina Đukić1, Øyvind Strømland1,2, Jonas Damgaard Elsborg3

  • 1Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, UK.

Science Advances
|September 13, 2023
PubMed

Insights

Poly (ADP-ribose) polymerase 14 (PARP14) is a dual-function enzyme with hydrolase activity. Its macrodomain 1 reverses PARP14 ADP-ribosylation, impacting cellular processes and antiviral responses.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Poly (ADP-ribose) polymerase 14 (PARP14) regulates immunity, transcription, and DNA replication stress.
  • The enzymatic activities and substrates of PARP14, particularly its ADP-ribosylation reversal mechanisms, remain largely uncharacterized.

Purpose of the Study:

  • To elucidate the enzymatic functions of PARP14, focusing on its ADP-ribosyl transferase and hydrolase activities.
  • To identify the substrates and regulatory mechanisms of PARP14-mediated ADP-ribosylation.
  • To investigate the role of PARP14's macrodomain 1 in its enzymatic activity and cellular function.

Main Methods:

  • Biochemical assays to assess ADP-ribosyl transferase and hydrolase activities of PARP14 and its mutants.
  • Cellular expression of PARP14 mutants to analyze in vivo ADP-ribosylation levels and cellular phenotypes.
  • In vitro and cellular assays to evaluate the interaction and activity of SARS-CoV-2 Nsp3 Mac1 domain on PARP14.

Main Results:

  • PARP14 exhibits dual enzymatic functions, acting as both an ADP-ribosyl transferase and a hydrolase on protein and nucleic acid substrates.
  • PARP14's macrodomain 1 possesses active ADP-ribosyl hydrolase activity, as does the macrodomain of PARP9.
  • Inactivation of macrodomain 1 in PARP14 leads to increased protein mono(ADP-ribosyl)ation in human cells, affecting PARP14, PARP13, and inducing specific cellular phenotypes.
  • The SARS-CoV-2 Nsp3 Mac1 domain efficiently reverses PARP14 ADP-ribosylation in vitro and in cells.

Conclusions:

  • PARP14 is a bifunctional enzyme critical for regulating cellular ADP-ribosylation.
  • The hydrolase activity of PARP14's macrodomain 1 plays a key role in reversing its own ADP-ribosylation.
  • Viral macrodomains, like SARS-CoV-2 Nsp3 Mac1, can counteract host PARP14-mediated antiviral responses, suggesting an evolutionary adaptation.

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