A protein-protein interaction underlies the molecular basis for substrate recognition by an adenosine-to-inosine RNA-editing enzyme

Nucleic Acids Res. 2018 Oct 12;46(18):9647-9659. doi: 10.1093/nar/gky800.

Abstract

Adenosine deaminases that act on RNA (ADARs) convert adenosine to inosine within double-stranded regions of RNA, resulting in increased transcriptomic diversity, as well as protection of cellular double-stranded RNA (dsRNA) from silencing and improper immune activation. The presence of dsRNA-binding domains (dsRBDs) in all ADARs suggests these domains are important for substrate recognition; however, the role of dsRBDs in vivo remains largely unknown. Herein, our studies indicate the Caenorhabditis elegans ADAR enzyme, ADR-2, has low affinity for dsRNA, but interacts with ADR-1, an editing-deficient member of the ADAR family, which has a 100-fold higher affinity for dsRNA. ADR-1 uses one dsRBD to physically interact with ADR-2 and a second dsRBD to bind to dsRNAs, thereby tethering ADR-2 to substrates. ADR-2 interacts with >1200 transcripts in vivo, and ADR-1 is required for 80% of these interactions. Our results identify a novel mode of substrate recognition for ADAR enzymes and indicate that protein-protein interactions can guide substrate recognition for RNA editors.

Publication types

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

MeSH terms

  • Adenosine / metabolism
  • Adenosine Deaminase / genetics*
  • Adenosine Deaminase / metabolism
  • Animals
  • Animals, Genetically Modified
  • Binding, Competitive
  • Caenorhabditis elegans / enzymology
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Deamination
  • Gene Expression Profiling
  • Inosine / metabolism
  • Mutation
  • Protein Binding
  • RNA Editing*
  • RNA, Double-Stranded / genetics*
  • RNA, Double-Stranded / metabolism
  • Substrate Specificity

Substances

  • Caenorhabditis elegans Proteins
  • RNA, Double-Stranded
  • Inosine
  • Adenosine Deaminase
  • Adr-1 protein, C elegans
  • Adr-2 protein, C elegans
  • Adenosine