Structural and biochemical studies of the 5'→3' exoribonuclease Xrn1

Nat Struct Mol Biol. 2011 Mar;18(3):270-6. doi: 10.1038/nsmb.1984. Epub 2011 Feb 6.

Abstract

The 5'→3' exoribonucleases (XRNs) have important functions in transcription, RNA metabolism and RNA interference. The structure of Rat1 (also known as Xrn2) showed that the two highly conserved regions of XRNs form a single, large domain that defines the active site of the enzyme. Xrn1 has a 510-residue segment after the conserved regions that is required for activity but is absent from Rat1/Xrn2. Here we report the crystal structures of Kluyveromyces lactis Xrn1 (residues 1-1,245, E178Q mutant), alone and in complex with a Mn(2+) ion in the active site. The 510-residue segment contains four domains (D1-D4), located far from the active site. Our mutagenesis and biochemical studies show that their functional importance results from their ability to stabilize the conformation of the N-terminal segment of Xrn1. These domains might also constitute a platform that interacts with protein partners of Xrn1.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Catalytic Domain
  • Exoribonucleases / chemistry*
  • Exoribonucleases / genetics
  • Exoribonucleases / metabolism
  • Kluyveromyces / chemistry
  • Kluyveromyces / enzymology*
  • Kluyveromyces / genetics
  • Kluyveromyces / metabolism
  • Manganese / chemistry
  • Manganese / metabolism*
  • Models, Molecular
  • Mutation
  • Protein Structure, Tertiary

Substances

  • Manganese
  • Exoribonucleases