Protein synthesis, amino acid uptake, membrane potential, cell volume, Na+ and K+ levels, and ATPase (Na+,K+ activated; EC 220.127.116.11) activity were investigated in control and poliovirus-infected HeLa cells. Inhibition of protein synthesis was first observed 60 min postinfection and reached a maximum at 120 min. The onset of protein synthesis inhibition coincided with a decrease in cell volume and with an elevation of ATPase activity in isolated HeLa cell membranes. Some 3 h after virus adsorption, ATPase activity was inhibited, the Na+-K+ gradient of the cell collapsed, both membrane potential-dependent tetraphenylphosphonium ion uptake and amino acid uptake were reduced, and the cell volume increased. These results provide further experimental support for the hypothesis that modification of the cell membrane plays an important role in the strategy of cytopathogenic viruses in the shutoff of host metabolism and cell death.