title: Macrodomain-containing proteins are new mono-ADP-ribosylhydrolases.
abstract: ADP-ribosylation is an important post-translational protein modification (PTM) that regulates diverse biological processes. ADP-ribosyltransferase diphtheria toxin-like 10 (ARTD10, also known as PARP10) mono-ADP-ribosylates acidic side chains and is one of eighteen ADP-ribosyltransferases that catalyze mono- or poly-ADP-ribosylation of target proteins. Currently, no enzyme is known that reverses ARTD10-catalyzed mono-ADP-ribosylation. Here we report that ARTD10-modified targets are substrates for the macrodomain proteins MacroD1, MacroD2 and C6orf130 from Homo sapiens as well as for the macrodomain protein Af1521 from archaebacteria. Structural modeling and mutagenesis of MacroD1 and MacroD2 revealed a common core structure with Asp102 and His106 of MacroD2 implicated in the hydrolytic reaction. Notably, MacroD2 reversed the ARTD10-catalyzed, mono-ADP-ribose-mediated inhibition of glycogen synthase kinase 3β (GSK3β) in vitro and in cells, thus underlining the physiological and regulatory importance of mono-ADP-ribosylhydrolase activity. Our results establish macrodomain-containing proteins as mono-ADP-ribosylhydrolases and define a class of enzymes that renders mono-ADP-ribosylation a reversible modification.
authors.affiliation: Institute of Veterinary Biochemistry and Molecular Biology, University of Zurich, Zurich, Switzerland.
authors.forename: Florian
authors.initials: F
authors.lastname: Rosenthal
authors.forename: Karla L H
authors.initials: KL
authors.lastname: Feijs
authors.forename: Emilie
authors.initials: E
authors.lastname: Frugier
authors.forename: Mario
authors.initials: M
authors.lastname: Bonalli
authors.forename: Alexandra H
authors.initials: AH
authors.lastname: Forst
authors.forename: Ralph
authors.initials: R
authors.lastname: Imhof
authors.forename: Hans C
authors.initials: HC
authors.lastname: Winkler
authors.forename: David
authors.initials: D
authors.lastname: Fischer
authors.forename: Amedeo
authors.initials: A
authors.lastname: Caflisch
authors.forename: Paul O
authors.initials: PO
authors.lastname: Hassa
authors.forename: Bernhard
authors.initials: B
authors.lastname: Lüscher
authors.forename: Michael O
authors.initials: MO
authors.lastname: Hottiger
mesh_terms: D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D009699:N-Glycosyl Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000246:Adenosine Diphosphate Ribose; D009699:N-Glycosyl Hydrolases; C046422:ADP-ribosylarginine hydrolase