Solution structure of the RWD domain of the mouse GCN2 protein

Protein Sci. 2004 Aug;13(8):2089-100. doi: 10.1110/ps.04751804.

Abstract

GCN2 is the alpha-subunit of the only translation initiation factor (eIF2alpha) kinase that appears in all eukaryotes. Its function requires an interaction with GCN1 via the domain at its N-terminus, which is termed the RWD domain after three major RWD-containing proteins: RING finger-containing proteins, WD-repeat-containing proteins, and yeast DEAD (DEXD)-like helicases. In this study, we determined the solution structure of the mouse GCN2 RWD domain using NMR spectroscopy. The structure forms an alpha + beta sandwich fold consisting of two layers: a four-stranded antiparallel beta-sheet, and three side-by-side alpha-helices, with an alphabetabetabetabetaalphaalpha topology. A characteristic YPXXXP motif, which always occurs in RWD domains, forms a stable loop including three consecutive beta-turns that overlap with each other by two residues (triple beta-turn). As putative binding sites with GCN1, a structure-based alignment allowed the identification of several surface residues in alpha-helix 3 that are characteristic of the GCN2 RWD domains. Despite the apparent absence of sequence similarity, the RWD structure significantly resembles that of ubiquitin-conjugating enzymes (E2s), with most of the structural differences in the region connecting beta-strand 4 and alpha-helix 3. The structural architecture, including the triple beta-turn, is fundamentally common among various RWD domains and E2s, but most of the surface residues on the structure vary. Thus, it appears that the RWD domain is a novel structural domain for protein-binding that plays specific roles in individual RWD-containing proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Mice
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Kinases / chemistry*
  • Protein Serine-Threonine Kinases
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sequence Alignment*
  • Structural Homology, Protein*
  • eIF-2 Kinase / chemistry

Substances

  • Protein Kinases
  • Eif2ak4 protein, mouse
  • Protein Serine-Threonine Kinases
  • eIF-2 Kinase

Associated data

  • PDB/1UKX