Inhibition of vaccinia virus late protein synthesis by isatin-β-thiosemicarbazone: Characterization and in vitro translation of viral mRNA

Thespecific effect of istin-βthiosemicarbozone (IBT) was manifested after vaccinia virus late protein synthesis had commenced. At 6 hr after infection, viral protein synthesis was inhibited by about 9596. We confirmed that λ portion of the virus-specific RNA appears to be degraded (B. Woodson and W. K. Joklik, 1965, Proc. Nat. Acad. Sci. USA 54,946-953). Nevertheless, the amount of viral RNA that was capped, properly methylated, and polyadenylylated, was reduced by only about 50%. Moreover, RNA from IBT-treated cells stimulated cell-free protein synthesis to one-half the level obtained with RNA from control cells. Polyacrylamide gel electrophoretic analysis further demonstrated that RNA from IBT-treated cells was translated into late viral proteins in vitro. Thus, it seems possible that the inhibition of protein synthesis in IBT-treated cells does not result entirely or directly from either an inhibition of mRNA synthesis or from λ depletion of mRNA caused by accelerated degradation. An alternative possibility, that accelerated degradation is secondary to λ more immediate effect of the drug on protein synthesis, was considered.