1. Recognition of mono-ADP-ribosylated ARTD10 substrates by ARTD8 macrodomains
- Author
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Tobias Karlberg, Henning Kleine, Petri Kursula, Annika Gross, Karla L. H. Feijs, Nicolas Herzog, Elisabeth Kremmer, Andreas G. Ladurner, Bernhard Lüscher, Patricia Verheugd, Bianca Nijmeijer, Herwig Schüler, Alexandra H. Forst, and Ann-Gerd Thorsell
- Subjects
Protein domain ,Plasma protein binding ,Molecular Dynamics Simulation ,Biology ,Crystallography, X-Ray ,Protein Structure, Secondary ,Histones ,Mice ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Protein structure ,Structural Biology ,Escherichia coli ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Binding site ,Molecular Biology ,030304 developmental biology ,ADP Ribose Transferases ,Adenosine Diphosphate Ribose ,0303 health sciences ,Binding Sites ,HEK 293 cells ,Isothermal titration calorimetry ,Recombinant Proteins ,Isoenzymes ,Molecular Docking Simulation ,Kinetics ,HEK293 Cells ,ran GTP-Binding Protein ,Biochemistry ,030220 oncology & carcinogenesis ,Mutation ,Ran ,Biophysics ,Thermodynamics ,NAD+ kinase ,Protein Binding - Abstract
Summary 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.
- Published
- 2013