Pathway of vesicular stomatitis virus entry leading to infection
- 1 April 1982
- journal article
- research article
- Published by Elsevier BV in Journal of Molecular Biology
- Vol. 156 (3), 609-631
- https://doi.org/10.1016/0022-2836(82)90269-8
Abstract
The entry of vesicular stomatitis virus into Madin-Darby canine kidney (MDCK) cells was examined both biochemically and morphologically. At low multiplicity and 0 °C, viruses bound to the cell surface but were not internalized. Binding was very dependent on pH. More than ten times more virus bound at pH 6.5 than at higher pH values. At the optimal pH, binding failed to reach equilibrium after more than two hours. The proportion of virus bound was irreproducible and low, relative to the binding of other enveloped viruses. Over 90% of the bound viruses were removed by proteases. When cells with pre-bound virus were warmed to 37 °C, a proportion of the bound virus became protease-resistant with a half-time of about 30 minutes. After a brief lag period, degraded viral material was released into the medium. The protease-resistant virus was capable of infecting the cells and probably did so by an intracellular route, since ammonium chloride blocked the infection and slightly reduced the degradation of viral protein. When the entry process was observed by electron microscopy, viruses were seen bound to the cell surface at 0 °C and, after warming at 37 °C, within coated pits, coated vesicles and larger, smooth-surfaced vesicles. No fusion of the virus with the plasma membrane was observed at pH 7.4. When pre-bound virus was incubated at a pH below 6 for 30 seconds at 37 °C, about 40 to 50% of the pre-bound virus became protease-resistant. On the basis of this result and previously published experiments (White et al., 1981), it was concluded that vesicular stomatitis virus fuses to the MDCK cell plasma membrane at low pH. These experiments suggest that vesicular stomatitis virus enters MDCK cells by endocytosis in coated pits and coated vesicles, and is transported to the lysosome where the low pH triggers a fusion reaction ultimately leading to the transfer of the genome into the cytoplasm. The entry pathway of vesicular stomatitis virus thus resembles that described earlier for both Semliki Forest virus and fowl plague virus.This publication has 51 references indexed in Scilit:
- The entry of diphtheria toxin into the mammalian cell cytoplasm: evidence for lysosomal involvement.The Journal of cell biology, 1980
- Polarized monolayers formed by epithelial cells on a permeable and translucent supportThe Journal of cell biology, 1978
- Role of the coated endocytic vesicle in the uptake of receptor-bound low density lipoprotein in human fibroblastsCell, 1977
- Immunofluorescence using dichlorotriazinylaminofluorescein (DTAF) I. Preparation and fractionation of labelled IgGJournal of Immunological Methods, 1976
- Lysosomotropic agentsBiochemical Pharmacology, 1974
- Electron Microscopic Observations on the Entry of Influenza Virus into Susceptible CellsJournal of General Virology, 1974
- Quantitative electron microscopic analysis of the penetration of VSV into L cellsVirology, 1974
- Early events in cell-animal virus interactionsMicrobiology and Molecular Biology Reviews, 1973
- Conjugates of Immunoglobulin G with Different Fluorochromes. I. Characterization by Anionic‐exchange ChromatographyScandinavian Journal of Immunology, 1973
- Buffer Combinations for Mammalian Cell CultureScience, 1971