Effects of monovalent cations on Semliki Forest virus entry into BHK-21 cells.
Infection of mammalian cells with Semliki Forest virus requires the endocytosis of the virus, its delivery to prelysosomal endosomes, and fusion of the viral envelope with the endosome membrane. Previous studies have indicated that the low endosomal pH triggers a conformational change in the viral spike glycoproteins rendering them fusogenic. In this paper, we demonstrate an additional factor(s) which regulates virus fusion in endosomes. We found that Semliki Forest virus is unable to penetrate or infect baby hamster kidney (BHK-21) cells grown in medium containing reduced Na+ concentrations. Virus endocytosis and degradation are nearly normal, the virus is transported to endosomes where a characteristic low pH-induced loss of trypsin-sensitivity of the E1 spike glycoprotein occurs. Nevertheless, the viral envelope fails to fuse with the endosomal membrane and the viral RNA is not released into the cytosol. As judged by the uptake of the voltage-sensitive probe [3H]triphenylmethyl phosphonium we observed a close correlation between conditions which inhibit virus infection and which cause depolarization of the cells. We propose that in intact cells, the fusion of Semliki Forest virus with the endosome membrane depends not only on acidic endosomal pH, but also on the maintenance of the potential.
D000818:Animals; D002414:Cations, Monovalent / Q000494:pharmacology*; D002460:Cell Line; D002794:Choline / Q000494:pharmacology; D006224:Cricetinae; D004705:Endocytosis; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000166:cytology; D008081:Liposomes / Q000378:metabolism; D008247:Lysosomes / Q000032:analysis; D008564:Membrane Potentials; D008854:Microscopy, Electron; D011188:Potassium / Q000494:pharmacology; D012367:RNA, Viral / Q000032:analysis; D012260:Ribonucleases / Q000378:metabolism; D012672:Semliki forest virus / Q000378:metabolism / Q000472:pathogenicity*; D012964:Sodium / Q000494:pharmacology; D014634:Valinomycin / Q000494:pharmacology
D002414:Cations, Monovalent; D008081:Liposomes; D012367:RNA, Viral; D014634:Valinomycin; D012964:Sodium; D012260:Ribonucleases; D002794:Choline; D011188:Potassium