Evidence that plasma membrane electrical potential is required for vesicular stomatitis virus infection of MDCK cells: a study using fluorescence measurements through polycarbonate supports.
We used fluorescence microscopy of Madin-Darby Canine Kidney (MDCK) cells grown on polycarbonate filters to study a possible link between plasma membrane electrical potential (delta psi pm) and infectivity of vesicular stomatitis virus (VSV). Complete substitution of K+ for extracellular Na+ blocks VSV infection of MDCK cells as well as baby hamster kidney (BHK) cells. When we independently perfused the apical and basal-lateral surfaces of high resistance monolayers, high K+ inhibited VSV infection of MDCK cells only when applied to the basal-lateral side; high K+ applied apically had no effect on VSV infection. This morphological specificity correlates with a large decrease in delta psi pm of MDCK cells when high K+ buffer is perfused across the basal-lateral surface. Depolarization of the plasma membrane by 130 mM basal K+ causes a sustained increase of cytosol pH in MDCK cells from 7.3 to 7.5 as reported by the fluorescent dye BCECF. Depolarization also causes a transient increase of cytosol Ca2+ from 70 to 300 nM as reported by the dye Fura-2. Neither increase could explain the block of VSV infectivity by plasma membrane depolarization. One alternative hypothesis is that delta psi pm facilitates membrane translocation of viral macromolecules as previously described for colicins, mitochondrial import proteins, and proteins secreted by Escherichia coli.
Akeson
M
M
Laboratory of Molecular Biology, National Institute of Mental Health, Bethesda, Maryland 20892.
Scharff
J
J
Sharp
C M
CM
Neville
D M
DM
D000818:Animals; D001464:Barium / Q000494:pharmacology; D017609:Barium Compounds; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D002712:Chlorides; D006224:Cricetinae; D004285:Dogs; D008564:Membrane Potentials; D008856:Microscopy, Fluorescence; D011076:Polycarboxylate Cement; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development / Q000502:physiology*; D010948:Viral Plaque Assay
D017609:Barium Compounds; D002712:Chlorides; D011076:Polycarboxylate Cement; C024986:barium chloride; D001464:Barium; C005506:polycarbonate; D012964:Sodium; D011188:Potassium; D002118:Calcium