Recognition of Mono-ADP-Ribosylated ARTD10 Substrates by ARTD8 Macrodomains

• Published in: Structure (London) Vol. 21; no. 3; pp. 462 - 475
• Main Authors: Forst, Alexandra H., Karlberg, Tobias, Herzog, Nicolas, Thorsell, Ann-Gerd, Gross, Annika, Feijs, Karla L.H., Verheugd, Patricia, Kursula, Petri, Nijmeijer, Bianca, Kremmer, Elisabeth, Kleine, Henning, Ladurner, Andreas G., Schüler, Herwig, Lüscher, Bernhard
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05-03-2013
• Subjects: Adaptors; Adenosine Diphosphate Ribose - chemistry; ADP Ribose Transferases - chemistry; ADP Ribose Transferases - genetics; Animals; Arts; Binding Sites; Crystal structure; Crystallography, X-Ray; Escherichia coli - genetics; HEK293 Cells; Histones - chemistry; Histones - genetics; Humans; Isoenzymes - chemistry; Isoenzymes - genetics; Kinetics; Medicin och hälsovetenskap; Mice; Molecular Docking Simulation; Molecular Dynamics Simulation; Mutation; Polymers; Protein Binding; Protein Interaction Domains and Motifs; Protein Structure, Secondary; Proteins; ran GTP-Binding Protein - chemistry; ran GTP-Binding Protein - genetics; Readers; Recognition; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Structure-Activity Relationship; Thermodynamics; Titration calorimetry
• ISSN: 0969-2126; 1878-4186; 1878-4186
• DOI: 10.1016/j.str.2012.12.019

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.
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► ARTD8 macrodomains read ARTD10-dependent mono-ADP-ribosylation ► The structure of ARTD8 macrodomains reveals a conserved fold for ADP-ribose binding ► Distinct macrodomains read mono- and poly-ADP-ribosylation selectively ► ARTD8 macrodomains can be used to visualize mono-ADP-ribosylation in cells
Frost et al. demonstrate that the ADP-ribosyltransferase ARTD10 mono-ADP-ribosylates target proteins, which are interaction motifs of ARTD8 macrodomains that interact with mono- but not poly-ADP-ribosylated substrates. This demonstrates mono-ADP-ribosylated proteins in cells and defines specific readers.