Reversible inhibition of induction of mengovirus RNA polymerase and of virus maturation in Novikoff rat hepatoma cells by phenethyl alcohol

Abstract

The addition of phenethyl alcohol (PEA) at 16–24 m M to suspension cultures of Novikoff rat hepatoma cells between 0.5 and 2.5 hours after infection with mengovirus completely prevents the induction of the viral RNA polymerase and the syntheses of virus-specific single-stranded and double-stranded RNA's. Each of these processes is inhibited about 80% by 12 m M PEA, but little affected by 4 m M. If cultures of infected cells are treated with 16 m M PEA at 0.5 hour after infection and the chemical is removed during the next hour, RNA polymerase activity appears in infected cells at the normal time (2.5–3 hours) and rate. If PEA is present between 1.5 and 2.5 hours after infection, on the other hand, the appearance of polymerase activity is delayed by 1 hour. Puromycin affects the formation of viral polymerase in a similar manner. It is concluded that the inhibition of RNA polymerase formation by PEA is a consequence of the PEA-induced inhibition of protein synthesis, and that the elicitation of viral RNA polymerase activity in infected cells requires continuous protein synthesis during the 1-hour period preceding the beginning of its appearance in the cell. The incorporation of viral RNA into virus particles is inhibited by PEA at concentrations (4–8 m M) which have little effect on the induction of viral RNA polymerase, the syntheses of virus-specific RNA's, or the synthesis of proteins by infected cells. Upon removal of PEA from the cells, virus maturation commences immediately at the normal rate.