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

• Published in: Nature communications Vol. 14; no. 1; pp. 3200 - 18
• Main Authors: Fontana, Pietro, Buch-Larsen, Sara C., Suyari, Osamu, Smith, Rebecca, Suskiewicz, Marcin J., Schützenhofer, Kira, Ariza, Antonio, Rack, Johannes Gregor Matthias, Nielsen, Michael L., Ahel, Ivan
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02.06.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 13/106; 13/89; 14/35; 631/1647/296; 631/181/2806; 631/337/1427/2566; 631/45/535/1266; 82/58; 82/83; Adenosine diphosphate; Adenosine Diphosphate Ribose - metabolism; ADP-Ribosylation; Animalia; Animals; Damage; Deoxyribonucleic acid; DNA; DNA damage; Drosophila; Drosophila - metabolism; Drosophila melanogaster - genetics; Drosophila melanogaster - metabolism; Evolutionary conservation; Fruit flies; Genomes; Humanities and Social Sciences; Insects; Life Sciences; Mammals; Mammals - metabolism; multidisciplinary; Organisms; Poly Adenosine Diphosphate Ribose - chemistry; Poly(ADP-ribose) polymerase; Polymers; Proteomics; Ribose; Ribosylation; Science; Science (multidisciplinary); Serine; Serine - metabolism; Signal transduction; Signaling; Wildlife conservation; DNA-DAMAGE, RESPONSEPOLY(ADP-RIBOSE) GLYCOHYDROLASE, CHROMATIN-STRUCTURE, POLYMERASE PARP, MODEL, MAINTENANCE, DOMAINS, SYSTEMS, TARGETRIBOSE
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-38793-y

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 d Parp1: d Hpf1 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. In the DNA damage response, ADP-ribosylation is an essential signaling pathway. Here the authors utilize a multidisciplinary approach to establish its molecular basis in fruit flies and provide evidence for Drosophila’s suitability as model organism.