An ADAR that edits transcripts encoding ion channel subunits functions as a dimer

EMBO J. 2003 Jul 1;22(13):3421-30. doi: 10.1093/emboj/cdg327.

Abstract

In this report, we establish that Drosophila ADAR (adenosine deaminase acting on RNA) forms a dimer on double-stranded (ds) RNA, a process essential for editing activity. The minimum region required for dimerization is the N-terminus and dsRNA-binding domain 1 (dsRBD1). Single point mutations within dsRBD1 abolish RNA-binding activity and dimer formation. These mutations and glycerol gradient analysis indicate that binding to dsRNA is important for dimerization. However, dimerization can be uncoupled from dsRNA-binding activity, as a deletion of the N-terminus (amino acids 1-46) yields a monomeric ADAR that retains the ability to bind dsRNA but is inactive in an editing assay, demonstrating that ADAR is only active as a dimer. Different isoforms of ADAR with different editing activities can form heterodimers and this can have a significant effect on editing in vitro as well as in vivo. We propose a model for ADAR dimerization whereby ADAR monomers first contact dsRNA; however, it is only when the second monomer binds and a dimer is formed that deamination occurs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Deaminase / chemistry
  • Adenosine Deaminase / genetics
  • Adenosine Deaminase / metabolism*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • DNA Primers
  • Dimerization
  • Drosophila
  • Ion Channels / genetics*
  • Molecular Sequence Data
  • Protein Binding
  • RNA Editing*
  • RNA, Messenger / genetics*
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • Ion Channels
  • RNA, Messenger
  • Adenosine Deaminase