title: Recognition of mono-ADP-ribosylated ARTD10 substrates by ARTD8 macrodomains.
abstract: ADP-ribosyltransferases (ARTs) catalyze the transfer of ADP-ribose from NAD(+) onto substrates. Some ARTs generate in an iterative process ADP-ribose polymers that serve as adaptors for distinct protein domains. Other ARTs, exemplified by ARTD10, function as mono-ADP-ribosyltransferases, but it has been unclear whether this modification occurs in cells and how it is read. We observed that ARTD10 colocalized with ARTD8 and defined its macrodomains 2 and 3 as readers of mono-ADP-ribosylation both in vitro and in cells. The crystal structures of these two ARTD8 macrodomains and isothermal titration calorimetry confirmed their interaction with ADP-ribose. These macrodomains recognized mono-ADP-ribosylated ARTD10, but not poly-ADP-ribosylated ARTD1. This distinguished them from the macrodomain of macroH2A1.1, which interacted with poly- but not mono-ADP-ribosylated substrates. Moreover, Ran, an ARTD10 substrate, was also read by ARTD8 macrodomains. This identifies readers of mono-ADP-ribosylated proteins, defines their structures, and demonstrates the presence of this modification in cells.
authors.affiliation: Institute of Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Aachen, Germany.
authors.forename: Alexandra H
authors.initials: AH
authors.lastname: Forst
authors.forename: Tobias
authors.initials: T
authors.lastname: Karlberg
authors.forename: Nicolas
authors.initials: N
authors.lastname: Herzog
authors.forename: Ann-Gerd
authors.initials: AG
authors.lastname: Thorsell
authors.forename: Annika
authors.initials: A
authors.lastname: Gross
authors.forename: Karla L H
authors.initials: KL
authors.lastname: Feijs
authors.forename: Patricia
authors.initials: P
authors.lastname: Verheugd
authors.forename: Petri
authors.initials: P
authors.lastname: Kursula
authors.forename: Bianca
authors.initials: B
authors.lastname: Nijmeijer
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Kremmer
authors.forename: Henning
authors.initials: H
authors.lastname: Kleine
authors.forename: Andreas G
authors.initials: AG
authors.lastname: Ladurner
authors.forename: Herwig
authors.initials: H
authors.lastname: Schüler
authors.forename: Bernhard
authors.initials: B
authors.lastname: Lüscher
mesh_terms: D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000235:genetics; D000246:Adenosine Diphosphate Ribose / Q000737:chemistry*; D000818:Animals; D001665:Binding Sites; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics; D057809:HEK293 Cells; D006657:Histones / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D051379:Mice; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D009154:Mutation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D020931:ran GTP-Binding Protein / Q000737:chemistry* / Q000235:genetics
chemical_list: D006657:Histones; D007527:Isoenzymes; D011994:Recombinant Proteins; C120756:macroH2A histone; D000246:Adenosine Diphosphate Ribose; D036002:ADP Ribose Transferases; D020931:ran GTP-Binding Protein