Mechanism of entry into the cytosol of poliovirus type 1: requirement for low pH.

Abstract

The effect of a number of drugs and culture conditions on the entry into human cervical carcinoma HeLa S3 cells of a strain of poliovirus 1 (Brunende) was tested. The cells were exposed in the dark to light-sensitive, neutral red-containing virus, in the presence of the drug to be tested. Then the cells were exposed to light, transferred to normal medium and incubated overnight. Cytopathogenic effect was measured as inhibition of [3H]leucine incorporation. Compounds that dissipate proton gradients across membranes, like monensin, protonophores and amines, and compounds that inhibit the acidification process, such as N,N''-dicyclohexylcarbodiimide (DCCD) and tributyltin, inhibited the entry of virus, but not virus binding. This was also the case with metabolic inhibitors that deplete cells for ATP. The same compounds also inhibited the cell-induced alteration of the virus particles. When cells with surface-bound virus were exposed to low pH, the virus entered efficiently, even in the presence of monensin and DCCD. Evidently, acidification somehow facilitates the entry of the virus RNA into the cytosol and under normal conditions the entry occurs from intracellular acidic vesicles.