Serine ADP-ribosylation in Drosophila provides insights into the evolution of reversible ADP-ribosylation signalling

  • Nat Commun. 2023 Jun 2;14(1):3200. doi: 10.1038/s41467-023-38793-y.
Pietro Fontana  #  1  2  3 ,  Sara C Buch-Larsen  #  4 ,  Osamu Suyari  #  1 ,  Rebecca Smith  1 ,  Marcin J Suskiewicz  1  5 ,  Kira Schützenhofer  1 ,  Antonio Ariza  6  7 ,  Johannes Gregor Matthias Rack  8  9 ,  Michael L Nielsen  10 ,  Ivan Ahel  11
Affiliations
  • 1. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
  • 2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
  • 3. Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
  • 4. Proteomics program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
  • 5. Centre de Biophysique Moléculaire, UPR4301 CNRS, rue Charles Sadron, CEDEX 2, F-45071, Orléans, France.
  • 6. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK. [email protected].
  • 7. School of Biosciences, University of Sheffield, Western Bank, Sheffield, S10 2TN, UK. [email protected].
  • 8. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK. [email protected].
  • 9. MRC Centre for Medical Mycology, School of Biosciences, University of Exeter, Geoffrey Pope Building, Exeter, EX4 4QD, UK. [email protected].
  • 10. Proteomics program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark. [email protected].
  • 11. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK. [email protected].
  • # Contributed equally.
PMID: 37268618 DOI: 10.1038/s41467-023-38793-y
Abstract

In the mammalian DNA damage response, ADP-ribosylation signalling is of crucial importance to MARK sites of DNA damage as well as recruit and regulate repairs factors. Specifically, the PARP1:HPF1 complex recognises damaged DNA and catalyses the formation of serine-linked ADP-ribosylation marks (mono-Ser-ADPr), which are extended into ADP-ribose Polymers (poly-Ser-ADPr) by PARP1 alone. Poly-Ser-ADPr is reversed by PARG, while the terminal mono-Ser-ADPr is removed by ARH3. Despite its significance and apparent evolutionary conservation, little is known about ADP-ribosylation signalling in non-mammalian Animalia. The presence of HPF1, but absence of ARH3, in some insect genomes, including Drosophila species, raises questions regarding the existence and reversal of serine-ADP-ribosylation in these species. Here we show by quantitative proteomics that Ser-ADPr is the major form of ADP-ribosylation in the DNA damage response of Drosophila melanogaster and is dependent on the dParp1:dHpf1 complex. Moreover, our structural and biochemical investigations uncover the mechanism of mono-Ser-ADPr removal by Drosophila Parg. Collectively, our data reveal PARP:HPF1-mediated Ser-ADPr as a defining feature of the DDR in Animalia. The striking conservation within this kingdom suggests that organisms that carry only a core set of ADP-ribosyl metabolising Enzymes, such as Drosophila, are valuable model organisms to study the physiological role of Ser-ADPr signalling.