Structure and specific RNA binding of ADAR2 double-stranded RNA binding motifs

Structure. 2006 Feb;14(2):345-55. doi: 10.1016/j.str.2005.11.013.

Abstract

Adenosine deaminases that act on RNA (ADARs) site-selectively modify adenosines to inosines within RNA transcripts, thereby recoding genomic information. How ADARs select specific adenosine moieties for deamination is poorly understood. Here, we report NMR structures of the two double-stranded RNA binding motifs (dsRBMs) of rat ADAR2 and an NMR chemical shift perturbation study of the interaction of the two dsRBMs with a 71 nucleotide RNA encoding the R/G site of the GluR-B. We have identified the protein and the RNA surfaces involved in complex formation, allowing us to present an NMR-based model of the complex. We have found that dsRBM1 recognizes a conserved pentaloop, whereas dsRBM2 recognizes two bulged bases adjacent to the editing site, demonstrating RNA structure-dependent recognition by the ADAR2 dsRBMs. In vitro mutagenesis studies with both the protein and the RNA further support our structural findings.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Deaminase / chemistry*
  • Adenosine Deaminase / metabolism
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Dimerization
  • Humans
  • Models, Molecular*
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular
  • Nucleic Acid Conformation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • RNA Editing
  • RNA, Double-Stranded / chemistry*
  • RNA, Double-Stranded / metabolism
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / metabolism
  • Rats
  • Sequence Alignment

Substances

  • RNA, Double-Stranded
  • RNA-Binding Proteins
  • ADARB1 protein, human
  • Adenosine Deaminase

Associated data

  • PDB/2B7T
  • PDB/2B7V