A lysine substitution in the ATP-binding site of eucaryotic initiation factor 4A abrogates nucleotide-binding activity

Mol Cell Biol. 1989 Sep;9(9):4061-3. doi: 10.1128/mcb.9.9.4061-4063.1989.

Abstract

Eucaryotic initiation factor 4A (eIF-4A) is a member of a family of proteins believed to be involved in the ATP-dependent melting of RNA secondary structure. These proteins contain a derivative of the consensus ATP-binding site AXXGXGKT. To assess the importance of the consensus amino acid sequence in eIF-4A for ATP binding, we mutated the consensus amino-proximal glycine and lysine to isoleucine and asparagine, respectively. The effect of the mutations was examined by UV-induced cross-linking of [alpha-32P]dATP to eIF-4A. Mutation of the lysine residue (but not of the glycine residue) resulted in the loss of [alpha-32P]dATP cross-linking to eIF-4A, suggesting that the lysine is an important determinant in ATP binding to eIF-4A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Eukaryotic Initiation Factor-4A
  • In Vitro Techniques
  • Lysine
  • Mice
  • Molecular Sequence Data
  • Mutation
  • Nucleotides / metabolism
  • Peptide Initiation Factors / genetics
  • Peptide Initiation Factors / metabolism*

Substances

  • Nucleotides
  • Peptide Initiation Factors
  • Adenosine Triphosphate
  • Eukaryotic Initiation Factor-4A
  • Lysine