Abstract
We describe here a genetic approach to the analysis of host cell functions involved in determining permissiveness to mouse hepatitis virus (MHV). Using the chemical mutagen, ethyl methane sulfonate (EMS), mouse fibroblast cell mutants were generated which were selected for resistance to cell-killing by MHV. These mutants were then screened for their susceptibility to MHV infection, ability to replicate MHV and relative sensitivity to MHV-induced cell fusion. In contrast to wild type L-2 cells which were acutely and terminally infected by MHV, all five mutants examined replicated MHV in a persistent manner. These mutants showed a reduced susceptibility to MHV infection and an increased resistance to MHV-induced cell fusion. Fusion resistance was specific to that mediated by the MHV E2 protein; mutant as well as wild type L-2 cells were equally sensitive to fusion by polyethylene glycol. The combined effect of reduced infectability and increased fusion resistance was to limit MHV infection to only a small percentage of the total cells in culture, thereby permitting survival of both virus and cells. The observed high rate of generation of the cell mutants suggests that the conversion of a fully MHV-susceptible cell to a semi-resistant one (capable of supporting a persistent infection) is a fairly common event, possibly involving a single mutation.
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References
Friedman, D.I., Olson, E.R., Georgopoulos, C, Tilly, K., Herskowitz, I. and Banuett, F. (1984) Interactions of bacteriophage and host macromolecules in the growth of bacteriophage lambda. Microbiol. Rev. 48, 299–325.
Toyama, S., Toyama, S. and Uetake, H. (1977) Altered cell-fusion capacity in lines of KB cells resistant to Sendai virus-induced cytolysis. Virology 76, 503–515.
Tufaro, F., Snider, M.D. and McKnight, S.L (1987) Identification and characterization of a mouse cell mutant defective in the intracellular transport of glycoproteins. J. Cell Biol. 105, 647–657.
Hara, T., Hattorl, S. and Kawakita, M. (1989) Isolation and characterization of mouse FM3A cell mutants which are devoid of Newcastle Disease Virus receptors. J. Virol. 63, 182–188.
Kaplan, G., Levy, A. and Racaniello, V.R. (1989) Isolation and characterization of HeLa cell lines blocked at different steps in the poliovirus life cycle. J. Virol. 63, 43–51.
Virelizier, J.L and Allison, A.C. (1976) Correlation of persistent mouse hepatitis virus (MHV-3) infection with its effects on mouse macrophage cultures. Arch. Virol. 50, 279–285.
MacNaughton, M.R. and Patterson, S. (1980) Mouse hepatitis virus strain 3 infection of C57, A/Sn and A/J strain mice and their macrophages. Arch. Virol. 66,71–75.
Arnheiter, H., Baechi, T. and Haller, O. (1982) Adult mouse hepatocytes in primary monolayer culture express genetic resistance to mouse hepatitis virus type 3. J. Immunol. 129, 1275–1281.
Lamontagne, L.M. and Dupuy, J.M. (1984) Natural resistance of mice to mouse hepatitis virus type 3 infection is expressed in embryonic fibroblast cells. J. Gen. Virol. 65, 1165–1171.
Tardieu, M., Boespflug, O., Barbe, T. (1986) Selective tropism of a neurotropic.Coronavirus for ependymal cells, neurons and meningeal cells. J. Virol. 60, 574–582.
Boyle, J.F., Weismiller, D.G. and Holmes, K.V. (1987) Genetic resistance to mouse hepatitis virus correlates with absence of virus-binding activity on target tissues. J. Virol. 61, 185–189.
Van Dinter, S. and Flintoff, W.F. (1987) Rat glial C6 cells are defective in murine Coronavirus internalization. J. Gen. Virol. 68,1677–1685.
Kooi, C., Mizzen, L, Alderson, C., Daya, M. and Anderson, R. (1988) Early events of importance in determining host cell permissiveness to mouse hepatitis virus infection. J. Gen. Virol. 69, 1125–1135.
Rothfels, K.H., Axelrad, A.A., Siminovitch, L, McCulloch, E.A. and Parker, R.C. (1959) The origin of altered cell lines from mouse, monkey and man as indicated by chromosome and transplantation studies. Canad. Cancer Conf. 3, 189–214.
Kit, S., Dubbs, D.R., Piekarski, L.J., Hsu, T.C. (1963) Deletion of thymidine kinase activity from L cells resistant to bromodeoxyuridine. Exp. Cell Res. 31, 297–312.
Manaker, R.A., Piczak, C.V., Miller, A.A. and Stanton, M.F. (1961) A hepatitis virus complicating studies with mouse leukemia. J. Natl. Cancer Inst. 27, 29–44.
Lucas, A., Flintoff, W., Anderson, R., Percy, D., Coulter, M. and Dales, S. (1977) In vivo and in vitro models of demyelinating diseases: tropism of the JHM strain of murine hepatitis virus for cells of glial origin. Cell 12, 553–560.
Mizzen, L., Macintyre, G., Wong, F. and Anderson, R. (1987) Translational regulation in mouse hepatitis virus infection is not mediated by altered intracellular ion concentrations. J. Gen. Virol. 68, 2143–2151.
Mizzen, L, Cheley, S., Rao, M., Wolf, R. and Anderson, R. (1983) Fusion resistance and decreased infectability as major host cell determinants of Coronavirus persistence. Virology 128, 407–417.
Mizzen, L, Daya, M. and Anderson, R. (1987) The role of protease-dependent cell membrane fusion in persistent and lytic infections of murine hepatitis virus. Adv. Exp. Med. Biol. 218, 175–186.
Lucas, A., Coulter, M., Anderson, R., Dales, S. and Flintoff, W. (1978) In vivo and in vitro models of demyelinating diseases. II. Persistence and host-regulated thermosensitivity in cells of neural derivation infected with mouse hepatitis and measles viruses. Virology 88, 325–337.
Knobler, R.L., Tunison, L.A. and Oldstone, M.B.A. (1984) Host genetic control of mouse hepatitis virus type 4 (JHM strain) replication. I. Restriction of virus amplification and spread in macrophages from resistant mice. J. Gen. Virol. 65, 1543–1548.
Wilson, G.A.R. and Dales, S. (1988) In vivo and in vitro models of demyelinating disease: efficiency of virus spread and formation of infectious centers among glial cells is genetically determined by the murine host. J. Virol. 62, 3371–3377.
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© 1990 Plenum Press, New York
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Daya, M. et al. (1990). Mouse Fibroblast Mutants Selected for Survival Against Mouse Hepatitis Virus Infection Show Increased Resistance to Infection and Virus-Induced Cell Fusion. In: Cavanagh, D., Brown, T.D.K. (eds) Coronaviruses and their Diseases. Advances in Experimental Medicine and Biology, vol 276. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-5823-7_9
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DOI: https://doi.org/10.1007/978-1-4684-5823-7_9
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