Recognition of mono-ADP-ribosylated ARTD10 substrates by ARTD8 macrodomains

Structure. 2013 Mar 5;21(3):462-75. doi: 10.1016/j.str.2012.12.019.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / chemistry*
  • ADP Ribose Transferases / genetics
  • Adenosine Diphosphate Ribose / chemistry*
  • Animals
  • Binding Sites
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • HEK293 Cells
  • Histones / chemistry*
  • Histones / genetics
  • Humans
  • Isoenzymes / chemistry
  • Isoenzymes / genetics
  • Kinetics
  • Mice
  • Molecular Docking Simulation
  • Molecular Dynamics Simulation
  • Mutation
  • Protein Binding
  • Protein Interaction Domains and Motifs
  • Protein Structure, Secondary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Structure-Activity Relationship
  • Thermodynamics
  • ran GTP-Binding Protein / chemistry*
  • ran GTP-Binding Protein / genetics

Substances

  • Histones
  • Isoenzymes
  • Recombinant Proteins
  • macroH2A histone
  • Adenosine Diphosphate Ribose
  • ADP Ribose Transferases
  • ran GTP-Binding Protein

Associated data

  • PDB/3Q6Z
  • PDB/3Q71
  • PDB/3V2B
  • PDB/3VFQ
  • PDB/4ABK
  • PDB/4ABL
  • PDB/4D86