2. Academic Validation
  3. The regulatory landscape of the human HPF1- and ARH3-dependent ADP-ribosylome

The regulatory landscape of the human HPF1- and ARH3-dependent ADP-ribosylome

  • Nat Commun. 2021 Oct 8;12(1):5893. doi: 10.1038/s41467-021-26172-4.
Ivo A Hendriks # 1,  Sara C Buch-Larsen # 1,  Evgeniia Prokhorova 2,  Jonas D Elsborg 1,  Alexandra K L F S Rebak 1,  Kang Zhu 2,  Dragana Ahel 2,  Claudia Lukas 3,  Ivan Ahel 2,  Michael L Nielsen 4
Affiliations

Affiliations

  • 1 Proteomics Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
  • 2 Sir William Dunn School of Pathology, University of Oxford, Oxford, OX1 3RE, UK.
  • 3 Protein Signaling Program, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200, Copenhagen, Denmark.
  • 4 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].
  • # Contributed equally.
PMID: 34625544 DOI: 10.1038/s41467-021-26172-4
Abstract

Despite the involvement of Poly(ADP-ribose) polymerase-1 (PARP1) in many important biological pathways, the target residues of PARP1-mediated ADP-ribosylation remain ambiguous. To explicate the ADP-ribosylation regulome, we analyze human cells depleted for key regulators of PARP1 activity, histone PARylation factor 1 (HPF1) and ADP-ribosylhydrolase 3 (ARH3). Using quantitative proteomics, we characterize 1,596 ADP-ribosylation sites, displaying up to 1000-fold regulation across the investigated knockout cells. We find that HPF1 and ARH3 inversely and homogenously regulate the serine ADP-ribosylome on a proteome-wide scale with consistent adherence to lysine-serine-motifs, suggesting that targeting is independent of HPF1 and ARH3. Notably, we do not detect an HPF1-dependent target residue switch from serine to glutamate/aspartate under the investigated conditions. Our data support the notion that serine ADP-ribosylation mainly exists as mono-ADP-ribosylation in cells, and reveal a remarkable degree of histone co-modification with serine ADP-ribosylation and other post-translational modifications.

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