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Protein Synthesis and Membrane Integrity in Interferon-treated HeLa Cells Infected with Encephalomyocarditis Virus

Department of Microbiology, Centro de Biología Molecular, Universidad Autónoma, Canto Blanco, Madrid-34, Spain

The survival of interferon (IFN)-treated cells after encephalomyocarditis (EMC) virus infection depends on both the concentration of interferon and the multiplicity of infection (m.o.i.) used. The cell survived EMC infection if a high IFN/m.o.i. ratio was used in the experiment, whereas cell death took place at low IFN/m.o.i. ratios, even if IFN is also present during infection. Analysis by polyacrylamide gel electrophoresis of the proteins synthesized in IFN-treated cells subsequently infected with EMC indicated that no virus proteins were detected at either low or high multiplicities of infection. However, at low m.o.i. the cell survived and continued synthesizing cellular proteins exclusively, whereas at high m.o.i. a drastic shut-off of host protein synthesis took place. Virus which had been inactivated by u.v. irradiation was unable to cause the shut-off of host protein synthesis, either in control or in IFN-treated cells. This result suggests that some virus gene expression occurs in cells treated with IFN, although no virus protein synthesis was detected. The synthesis of virus RNA was also strongly inhibited after treatment of cells with IFN.

The integrity of the cell membrane in control and in IFN-treated cells was studied by analysing the ⁸⁶Rb⁺ ion leakage, the thymidine pool, the choline uptake and the entry of the translation inhibitor hygromycin B, to which cells are impermeable, at different times after EMC infection. The results obtained indicate that the early membrane leakiness observed after virus infection is not prevented by IFN treatment. However, the development of late leakiness to ⁸⁶Rb⁺ ions, thymidine and hygromycin B was not observed in IFN-treated cells.

Keywords: interferon, encephalomyocarditis virus, protein synthesis, membrane integrity

Received 9 March 1981; accepted 1 May 1981.