Structure and regulation of eukaryotic initiation factor eIF-2. Sequence of the site in the alpha subunit phosphorylated by the haem-controlled repressor and by the double-stranded RNA-activated inhibitor

Eur J Biochem. 1987 Jul 15;166(2):357-63. doi: 10.1111/j.1432-1033.1987.tb13523.x.

Abstract

Eukaryotic protein synthesis initiation factor 2 (eIF-2) can be phosphorylated on its alpha subunit by two well-characterised protein kinases, termed the haem-controlled repressor (HCR) and the double-stranded RNA-activated inhibitor (dsI). Phosphorylation of eIF-2 by these kinases is thought to be important in the regulation of peptide-chain initiation. We report the location of the serine residue in the alpha subunit, which is phosphorylated by both these enzymes. Limited tryptic digestion and subsequent cyanogen bromide treatment of rat liver eIF-2 phosphorylated by HCR yielded one major phosphopeptide. This peptide had the sequence Ile-Leu-Leu-Ser-Glu-Leu-Ser(P)-Arg-Arg. The same major phosphopeptide was obtained from rabbit reticulocyte eIF-2 phosphorylated by HCR or dsI as judged by its behaviour on two-dimensional mapping and reverse-phase chromatography. In all cases the phosphorylated residue was found to be serine-7, and not serine-4, of the above sequence as determined from sequence analysis and by subdigestion of the peptide with Staphylococcus aureus V8 proteinase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Eukaryotic Initiation Factor-2
  • Heme / pharmacology*
  • Liver / metabolism
  • Macromolecular Substances
  • Peptide Initiation Factors / isolation & purification
  • Peptide Initiation Factors / metabolism*
  • Phosphopeptides / isolation & purification
  • Phosphorylation
  • Proteins / isolation & purification
  • Proteins / metabolism*
  • RNA, Double-Stranded / metabolism*
  • Rabbits
  • Rats
  • Repressor Proteins / metabolism*
  • Reticulocytes / metabolism
  • Transcription Factors / metabolism*

Substances

  • Eukaryotic Initiation Factor-2
  • Macromolecular Substances
  • Peptide Initiation Factors
  • Phosphopeptides
  • Proteins
  • RNA, Double-Stranded
  • Repressor Proteins
  • Transcription Factors
  • Heme