Modification of membrane permeability during Semliki Forest virus infection

Modification of membrane permeability has been analyzed in Semliki Forest virus (SFV)-infected cells by means of translation inhibitors not permeable to normal cells. A higher inhibition of protein synthesis in the infected cells is only observed with those antibiotics that do not easily pass the cell membrane, but not with others, permeable to cells, such as anisomycin, cycloheximide, trichodermin, etc. It does not, therefore, seem that the suggestion of M. A. Gray, K. J. Micklem, and C. A. Pasternak [Eur. J. Biochem. 135, 299-302, (1983)] that protein synthesis in virus-infected cells is more susceptible to translation inhibitors in general is correct. Both low- and high-molecular weight compounds enter the cell very early during SFV infection. This permeabilization is blocked by compounds known to increase the pH of coated vesicles, such as NH4Cl and chloroquine. Inhibition of energy production by means of N3Na and 2'-deoxyglucose also blocks this process. The optimal external pH for this early permeabilization is around 7-8. Acidic pH inhibits the entry of these impermeant antibiotics promoted by SFV. Analysis of 86Rb+ content in SFV-infected HeLa cells also indicates that a drastic decline in this cation takes place, in agreement with previous findings, but disagreeing with the previous results. A parallel between the decrease in this cation and the blockade of protein synthesis is apparent, throughout the course of infection. In addition to the early permeabilization that takes place during virus entry, increased entry of hygromycin B and alpha-sarcin also occurs in SFV-infected cells from 2 to 3 hr postinfection, but not when late viral replication is blocked by means of interferon treatment.