Poliovirus-induced alterations in HeLa cell membrane functions

Abstract

Protein synthesis, amino acid uptake, membrane potential, cell volume, Na+ and K+ levels, and ATPase (Na+,K+ activated; EC 3.6.1.3) 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.

Evidence for poliovirus‐induced cytoplasmic alkalinization in Hela cells

“…The replication of many viruses in cell cultures is associated with alterations in cellular cation levels (reviewed in Nair, 1984). In HeLa cells infected with poliovirus, these changes first appear around 3 h postinfection (PI) and are characterized by Na + accumulation and K + leakage, resulting in part from the inhibition of Na+/Kf ATPase activity and in part from increased permeability of the plasma membrane (Nair et al, 1979;Nair, 1981;Schaefer et al, 1982). Between 1 and 1.5 h PI, when Na+/K+ contents are unaltered, cellular Na+/K+ ATPase activity is transiently stimulated in the absence of any increase in ATP content (Nair et al, 1979;Schaefer et al, 1982).…”

“…In HeLa cells infected with poliovirus, these changes first appear around 3 h postinfection (PI) and are characterized by Na + accumulation and K + leakage, resulting in part from the inhibition of Na+/Kf ATPase activity and in part from increased permeability of the plasma membrane (Nair et al, 1979;Nair, 1981;Schaefer et al, 1982). Between 1 and 1.5 h PI, when Na+/K+ contents are unaltered, cellular Na+/K+ ATPase activity is transiently stimulated in the absence of any increase in ATP content (Nair et al, 1979;Schaefer et al, 1982). In suspension cultures of HeLa cells, this virus-induced stimulation of Na+/K+ ATPase activity appears to coincide with a modest decrease in cell volume (Schaefer et al, 1982).…”

A reduction in the activity of the NA+, K+ ‐pump in dimethylsulfoxide‐treated friend erythroleukemia cells is not due to partial inactivation of the NA+, K+ ‐ATPase

“…Structural changes taking place in the membrane after DMSO addition mi h t be responsible for this transient increase in Na+, K -ATPase activity. A similar phenomenon could also be demonstrated in HeLa cells after infection with poliovirus (Schaefer et al, 1982). Whether this latent stimulation of the Na+, K+-ATPase has any functional significance or if it should be considered as an indication for structural changes after DMSO addition remains to be elucidated.…”

“…Na+-preloading of control and DMSO-induced cells was performed by the method of Pietrzyk et a1 (1978) with the alteration that incubations in ice-bath and 37°C were carried out in isotonic K'-free EBSS for 20 minutes. After this treatment, the concentration of Na+ increased and the concentration of K + decreased by at least 100 mM in cells as determined by methods described previously (Schaefer et al, 1982). "Rb+-uptake in Naf-preloaded cells was measured at elevated K f -concentration to provide more favorable conditions for the function of the Na+, K+-ATPase (Pietrzyk et al, 1978).…”

Poliovirus‐induced intracellular alkalinization involves a proton ATPase and protein phosphorylation

“…These alterations could have pathophysiological significance (Pasternak and Micklem, 1981;Nair, 1984). In poliovirus-infected HeLa cells, virus-induced early changes in plasma membrane (P)-mediated functions produce transient increases in Na ' /K ' ATPase activity (Nair et al, 1979;Schaefer et al, 1982) and membrane permeability (Carrasco et al, 1991) and a transient decrease in cellular volume (Schaefer et al, 1982). Later changes evidenced by inhibition of Na+/K+ ATPase activity (Nair et al, 1979;Schaefer et al, 19821, reciprocal alterations in intracellular K' and Na' (Nair, 1981;Schaefer et al, 19821, membrane depolarization (Schaefer et al, 1982), and leakiness (Nair et al, 1979;Schaefer et al, 1982;Lopez-Rivas et al, 1987) are irreversible.…”