Toxicity of diphtheria toxin-related proteins produced by suppression of nonsense mutations

The Mr = 62,000 diphtheria toxin-related proteins produced from the suppression of nonsense mutations within the tox gene of corynephage beta were purified by affinity chromatography. Except for the toxin 111-sup2-62, the Mr = 62,000 polypeptides were found to have the same specific toxicity as does wild type toxin. 111-sup2-62 was found to have a prolonged lag period prior to the onset of inhibition of protein synthesis and ADP-ribosylation of elongation factor 2. 111-sup2-62 differs from wild type toxin by an amino acid substitution at a site approximatley 47,000 daltons from the NH2 terminus. The data presented provide genetic support for the Boquet-Pappenheimer model (Boquet, P., and Pappenheimer, A. M. Jr (1978) J. Biol. Chem. 251, 5770-5778) of fragment A translocation into the eukaryotic cell cytosol.