Protein-Synthetic Activity of Ribosomes from Interferon-Treated Cells

Abstract

Cell-free protein-synthetic systems from normal and interferon-treated chick cells were compared. No difference was found in the amino acid incorporation activities of such ribosome-cell sap systems or in their response to polyuridylic acid. Throughout a variety of experiments we failed to detect the formation of a discrete peak of virus-specific polysomes, when ribosome monomers and subunits (from interferon-treated or control cells) were incubated with labeled Sindbis or Semliki Forest virus ribonucleic acid (RNA). Some binding of viral RNA did occur, but the complexes formed were evident in sucrose gradients as a broad, rapidly sedimenting shoulder on the ribosome monomer peak. Interferon pretreatment of cells did not affect the formation of these complexes in vitro, nor did it alter their rate of breakdown on incubation under amino acid incorporation conditions. Experiments with inhibitors of protein synthesis showed that such “breakdown” was not dependent upon amino acid incorporation and was not an index of translation. In these respects, our results are in marked contrast to those of Marcus and Salb. These results, together with our failure to detect any significant change in the protein composition of ribosomes from interferon-treated cells, suggest that such treatment does not result in a modification of the ribosome per se. They do not, however, rule out the involvement of a factor(s) required for ribosomes and viral RNA to function in viral protein synthesis. Indeed, it remains likely that interferon acts through such a mechanism, although the precise level at which the inhibition occurs remains to be elucidated.