2. Academic Validation
  3. Proteomic analyses identify ARH3 as a serine mono-ADP-ribosylhydrolase

Proteomic analyses identify ARH3 as a serine mono-ADP-ribosylhydrolase

  • Nat Commun. 2017 Dec 12;8(1):2055. doi: 10.1038/s41467-017-02253-1.
Jeannette Abplanalp 1 2,  Mario Leutert 1 2,  Emilie Frugier 3,  Kathrin Nowak 1 2,  Roxane Feurer 1,  Jiro Kato 4,  Hans V A Kistemaker 5,  Dmitri V Filippov 5,  Joel Moss 4,  Amedeo Caflisch 3,  Michael O Hottiger 6
Affiliations

Affiliations

  • 1 Department of Molecular Mechanisms of Disease, University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
  • 2 Molecular Life Science PhD Program of the Life Science Zurich Graduate School, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
  • 3 Department of Biochemistry, University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
  • 4 Laboratory of Translational Research, National Heart, Lung and Blood Institute, NIH, Bethesda, MD, 20892-1590, USA.
  • 5 Leiden Institute of Chemistry, Department of Bio-organic Synthesis, Leiden University, Einsteinweg 55, 2333 CC, Leiden, The Netherlands.
  • 6 Department of Molecular Mechanisms of Disease, University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland. [email protected].
PMID: 29234005 DOI: 10.1038/s41467-017-02253-1
Abstract

ADP-ribosylation is a posttranslational modification that exists in monomeric and polymeric forms. Whereas the writers (e.g. ARTD1/PARP1) and erasers (e.g. PARG, ARH3) of poly-ADP-ribosylation (PARylation) are relatively well described, the enzymes involved in mono-ADP-ribosylation (MARylation) have been less well investigated. While erasers for the MARylation of glutamate/aspartate and arginine have been identified, the respective enzymes with specificity for serine were missing. Here we report that, in vitro, ARH3 specifically binds and demodifies proteins and Peptides that are MARylated. Molecular modeling and site-directed mutagenesis of ARH3 revealed that numerous residues are critical for both the mono- and the poly-ADP-ribosylhydrolase activity of ARH3. Notably, a mass spectrometric approach showed that ARH3-deficient mouse embryonic fibroblasts are characterized by a specific increase in serine-ADP-ribosylation in vivo under untreated conditions as well as following hydrogen peroxide stress. Together, our results establish ARH3 as a serine mono-ADP-ribosylhydrolase and as an important regulator of the basal and stress-induced ADP-ribosylome.

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