Shiga Toxin from 'Shigella dysenteriae' 1 Inhibits Protein Synthesis in Reticulocyte Lysates by Inactivation of Aminoacyl-tRNA Binding

摘要

Inhibition of the peptide elongation cycle of eukaryotic protein synthesis by Shiga toxin from Shigella dysenteriae 1 was examined in toxin-treated reticulocyte lysate mixtures. Peptidyl transferase activity of toxin-treated ribosomes was measured by following the decrease in peptidyl-tRNA concentrations when puromycin was added after incubation with toxin. Concentrations of Tritium leucine-labeled peptidyl-tRNA were measured by extraction with cetyltrimethylammonium bromide. The data suggest that Shiga toxin inhibited aminoacyl-tRNA binding. Toxin-treated ribosomes retained peptidyl transferase activity, and toxin did not block translocation. Furthermore, no inhibition of initiation of protein synthesis could be observed. Finally, Shiga toxin had no detectable nucleolytic effect on polysomal 28S rRNA, nor was hydrolysis of 5.8S or 5S rRNA observed. (Reprints).