Abstract
Primary mouse glial cell cultures were infected with mouse hepatitis virus strain A59 (MHV-A59) and maintained over an 18 week period. Viruses isolated from these cultures 16–18 weeks postinfection produce small plaques on fibroblasts and cause only minimal levels of cell-to-cell fusion at times when wild type causes nearly complete cell fusion. However, when mutant-infected cultures were examined 24–36 hours postinfection approximately 90% of the cells were in syncytia showing that the fusion defect is not absolute but rather delayed. Addition of trypsin to mutant-infected cultures enhanced cell fusion a small (2- to 5-fold) but significant degree. Sequencing of portions of the spike genes of six fusion-defective mutants revealed that all contained the same single nucleotide mutation resulting in a substitution of aspartic acid for histidine in the spike cleavage signal. Mutant virions contained only the 180 kDa form of spike protein suggesting that this mutation prevented the normal proteolytic cleavage of the 180 kDa protein into the 90 kDa subunits. Examination of revertants of the mutants supports this hypothesis. Replacement of the negatively-charged aspartic acid with either the wild type histidine or a non-polar amino acid was associated with the restoration of spike protein cleavage and cell fusion.
Chapter PDF
Similar content being viewed by others
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
References
Spaan, W. J. M., Cavanagh, D., Horzinek, M. C. J. Gen. Virol. 69:2939–2952, 1988.
Gallagher, T. M., Escarmis, C., Buchmeier, M. J. J. Virol. 65:1916–1928, 1991.
Vennema, H., Heijne, L., Zijderveld, A, Horzinek, M. C., Spaan, W. J. M. J. Virol. 64:339–346, 1990.
Frana, M. F., Behnke, J. N., Sturman, S., Holmes, K. V. J. Virol. 56:912–920, 1985.
Luytjes, W., Sturman, L., Bredenbeck, P. J., Charite, J., van der Zeijst, B. A. M., Horzinek, M. C., Spaan, W. J. M. Virology 161:479–487, 1987.
Bosch, F. X., Garten, W., Klenk, H. D., Rott, R. Virology 113:725–735, 1981.
Glickman, R. L., Syddall, R. J., Iorio, R. M., Sheehan, J. P., Bratt, M. A. J. Virol. 62:354–356, 1988.
Perez, L. G., Hunter, E. J. Virol. 61:1609–1614, 1987.
Gombold, J.L., Weiss, S.R. Microb. Path. In Press.
Lavi, E., Suzumura, A., Hirayama, M., Highkin, M. K., Dambach, D. M., Silberberg, D. H.,Weiss, S. R. Microb. Path. 3:79–86, 1987.
Sawicki, S. G. Adv. Exp. Med. Biol. 218:169–174, 1987.
DeGroot, R. J., Luytjes, W., Horzinek, M. C., van der Zeijst, B. A. M., Spaan, W. J. M., Lenstra, J. A. J. Mol. Biol. 196:963–966, 1987.
Gallagher, T. M., Parker, S. E., Buchmeier, M. J. J. Virol. 64:731–741, 1990.
Parker, S.E., Gallagher, T. M., Buchmeier, M. J. Virology 173:664–673, 1989.
Morris, V.L., Tieszer, C., MacKinnon, J., Percy, D. Virology 169:127–136, 1989.
LaMonica, N., Banner, L., Morris, V. L., Lai, M. M. C. Virology 182:883–888, 1991.
Kawaoka, Y.,Webster, R. G. Proc. Natl. Acad. Sci. 85:324–328, 1988.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media New York
About this chapter
Cite this chapter
Gombold, J.L., Hingley, S.T., Weiss, S.R. (1994). Identification of Peplomer Cleavage Site Mutations Arising during Persistence of MHV-A59. In: Laude, H., Vautherot, JF. (eds) Coronaviruses. Advances in Experimental Medicine and Biology, vol 342. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2996-5_25
Download citation
DOI: https://doi.org/10.1007/978-1-4615-2996-5_25
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-6305-7
Online ISBN: 978-1-4615-2996-5
eBook Packages: Springer Book Archive