Expression of non-ADP-ribosylatable, diphtheria toxin-resistant elongation factor 2 in Saccharomyces cerevisiae

Biochem Biophys Res Commun. 1993 Mar 31;191(3):1145-51. doi: 10.1006/bbrc.1993.1336.


Eucaryotic elongation factor 2 (EF-2) contains a post-translationally modified histidine residue termed diphthamide that is specifically ADP-ribosylated by diphtheria toxin (DT) or Pseudomonas exotoxin A. To analyze the potential physiological role of ADP-ribosylation of EF-2 by cellular ADP-ribosyl transferase, we constructed DT-resistant, non-ADP-ribosylatable Saccharomyces cerevisiae EF-2 by site-directed mutagenesis and expressed the mutant EF-2 in yeast. Substitution of Arg for Gly(701) in yeast EF-2 conferred complete resistance to DT in vivo and in vitro. However, when only non-ribosylatable EF-2 was expressed in cells using genetic manipulation, the mutated EF-2 did not affect vegetative cell growth, mating, sporulation and germination of ascospores.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Diphosphate Ribose / metabolism*
  • Base Sequence
  • Cloning, Molecular
  • DNA Mutational Analysis
  • Diphtheria Toxin / toxicity*
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • Mutagenesis, Site-Directed
  • Oligodeoxyribonucleotides / chemistry
  • Peptide Elongation Factor 2
  • Peptide Elongation Factors / genetics
  • Peptide Elongation Factors / metabolism*
  • Saccharomyces cerevisiae / physiology*
  • Spores, Fungal


  • Diphtheria Toxin
  • Oligodeoxyribonucleotides
  • Peptide Elongation Factor 2
  • Peptide Elongation Factors
  • Adenosine Diphosphate Ribose