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. 2000 Feb 23;35(3):277–289. doi: 10.1016/0168-1702(94)00089-U

Molecular characterization of the S proteins of two enterotropic murine coronavirus strains

Satoshi Kunita a,1, Linong Zhang b, Felix R Homberger b, Susan R Compton a,∗
PMCID: PMC7134003  PMID: 7785316

Abstract

Enterotropic strains of murine coronaviruses (MHV-Y and MHV-RI) differ extensively in their pathogenesis from the prototypic respiratory strains of murine coronaviruses. In an effort to determine which viral proteins might be determinants of enterotropism, immunoblots of MHV-Y and MHV-RI virions using anti-S, -N and -M protein-specific antisera were performed. The uncleaved MHV-Y and MHV-RI S proteins migrated slightly faster than the MHV-A59 S protein. The MHV-Y S protein was inefficiently cleaved. The MHV-Y, MHV-RI and MHV-A59 N and M proteins showed only minor differences in their migration. The S genes of MHV-Y and MHV-RI were cloned, sequenced and found to encode 1361 and 1376 amino acid long proteins, respectively. The presence of several amino acids changes upstream from the predicted cleavage site of the MHV-Y S protein may contribute its inefficient cleavage. A high degree of homology was found between the MHV-RI and MHV-4 S proteins, whereas the homology between the MHV-Y S protein and the S proteins of other MHV strains was much lower. These results indicate that the enterotropism of MHV-RI and MHV-Y may be determined by different amino acid changes in the S protein and/or by changes in other viral proteins.

Keywords: Mouse hepatitis virus, Coronavirus, S glycoprotein

References

  1. Abraham S., Kienzle T.E., Lapps W., Brian D.A. Deduced sequence of the bovine coronavirus spike protein and identification of the internal proteolytic cleavage site. Virology. 1990;176:296–301. doi: 10.1016/0042-6822(90)90257-R. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armstrong J., Niemann H., Smeekens S., Rottier P., Warren G. Sequence and topology of a model intracellular membrane protein, E1 glycoprotein, from a coronavirus. Nature. 1984;308:751–752. doi: 10.1038/308751a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barthold S.W. In: Viral and Mycoplasmal Infections of Laboratory Rodents. Bhatt P.N., Jacoby R.O., editors. Academic Press; New York: 1986. Mouse hepatitis virus biology and epizootiology; pp. 571–601. [Google Scholar]
  4. Barthold S.W. Host age and genotypic effects on enterotropic mouse hepatitis virus infection. Lab. Anim. Sci. 1987;37:36–40. [PubMed] [Google Scholar]
  5. Barthold S.W., Smith A.L. Response of genetically susceptible and resistant mice to intranasal inoculation with mouse hepatitis virus JHM. Virus Res. 1987;7:225–239. doi: 10.1016/0168-1702(87)90030-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barthold S.W., Smith A.L., Lord P.F.S., Bhatt P.N., Jacoby R.O., Main A.J. Epizootic coronaviral typhlocolitis in suckling mice. Lab. Anim. Sci. 1982;32:376–383. [PubMed] [Google Scholar]
  7. Barthold S.W., Smith A.L., Povar M.L. Enterotropic mouse hepatitis virus infection in nude mice. Lab. Anim. Sci. 1985;35:613–618. [PubMed] [Google Scholar]
  8. Barthold S.W., Beck D.S., Smith A.L. Enterotropic coronavirus (mouse hepatitis virus) in mice: influence of host age and strain on infection and disease. Lab. Anim. Sci. 1993;43:276–284. [PubMed] [Google Scholar]
  9. Binns M.M., Boursnell M.E.G., Cavanaugh D., Pappin D.J.C., Brown T.D.K. Cloning and sequencing of the gene encoding the spike protein of the coronavirus IBV. J. Gen. Virol. 1985;66:719–726. doi: 10.1099/0022-1317-66-4-719. [DOI] [PubMed] [Google Scholar]
  10. Boireau P., Crucière C., LaPorte J. Nucleotide sequence of the glycoprotein S gene of bovine enteric coronavirus and comparison with the S proteins of two mouse hepatitis virus strains. J. Gen. Virol. 1990;71:487–492. doi: 10.1099/0022-1317-71-2-487. [DOI] [PubMed] [Google Scholar]
  11. Collins A.R., Knobler R.L., Powell H., Buchmeier M.J. Monoclonal antibodies to murine hepatitis virus 4 (strain JHM) define the viral glycoprotein responsible for attachment and cell fusion. Virology. 1982;119:358–371. doi: 10.1016/0042-6822(82)90095-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Compton S.R. Enterotropic strains of mouse coronavirus differ in their use of murine carcinoembryonic antigen-related glycoprotein receptors. Virology. 1994;203:197–201. doi: 10.1006/viro.1994.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Compton S.R., Barthold S.W., Smith A.L. The cellular and molecular pathogenesis of coronaviruses. Lab. Anim. Sci. 1993;43:15–28. [PubMed] [Google Scholar]
  14. Dalziel R.G., Lampert P.W., Talbot P.J., Buchmeier M.J. Site-specific alteration of murine hepatitis virus type 4 peplomer glycoprotein E2 results in reduced neurovirulence. J. Virol. 1986;59:463–471. doi: 10.1128/jvi.59.2.463-471.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. De Groot R.J., Maduro J., Lenstra J.A., Horzinek M.C., van der Zeijst B.A.M., Spaan W.J.M. cDNA cloning and sequence analysis of the gene encoding the peplomer protein of feline infectious peritonitis virus. J. Gen. Virol. 1987;68:2639–2646. doi: 10.1099/0022-1317-68-10-2639. [DOI] [PubMed] [Google Scholar]
  16. Dveksler G.S., Pensiero M.N., Cardellichio C.B., Williams R.K., Jiang G.-S., Holmes K.V., Dieffenbach C.W. Cloning of the mouse hepatitis virus (MHV) receptor: expression in human and hamster cell lines confers susceptibility to MHV. J. Virol. 1991;65:6881–6891. doi: 10.1128/jvi.65.12.6881-6891.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fazakerley J.K., Parker S.E., Bloom F., Buchmeier M.J. The V5A13.1 envelope glycoprotein deletion mutant of mouse hepatitis virus type-4 is neuroattenuated by its reduced rate of spread in the central nervous system. Virology. 1992;187:178–188. doi: 10.1016/0042-6822(92)90306-A. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fleming J.O., Trousdale M.D., El-Zaatari F.A.K., Stohlman S.A., Weiner L.A. Pathogenicity of antigenic variants of murine coronavirus JHM selected with monoclonal antibodies. J. Virol. 1986;58:869–875. doi: 10.1128/jvi.58.3.869-875.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Frana M.F., Behnke J.N., Sturman L.S., Holmes K.V. Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: host-dependent differences in proteolytic cleavage and cell fusion. J. Virol. 1985;56:912–920. doi: 10.1128/jvi.56.3.912-920.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gallagher T.M., Parker S.E., Buchmeier M.J. Neutralization-resistant variants of a neurotropic coronavirus are generated by deletions within the amino-terminal half of the spike glycoprotein. J. Virol. 1990;64:731–741. doi: 10.1128/jvi.64.2.731-741.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Genetics Computer Group, Inc. Genetics Computer Group, Inc; 575 Science Drive, Madison, WI 53711: 1991. (Genetics Computer Group Sequence Analysis Software Package Version 7.0 Program Manual). [Google Scholar]
  22. Gombold J.L., Hingley S.T., Weiss S.R. Fusion-defective mutants of mouse hepatitis virus A59 contain a mutation in the spike protein cleavage signal. J. Virol. 1993;67:4504–4512. doi: 10.1128/jvi.67.8.4504-4512.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hingley S.T., Gombold J.L., Lavi E., Weiss S.R. MHV-A59 fusion mutants are attenuated and display altered hepatotropism. Virology. 1994;200:1–10. doi: 10.1006/viro.1994.1156. [DOI] [PubMed] [Google Scholar]
  24. Homberger F.R. Nucleotide sequence comparison of membrane protein genes of three enterotropic strains of mouse hepatitis virus. Virus Res. 1994;31:49–56. doi: 10.1016/0168-1702(94)90070-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Homberger F.R., Smith A.L., Barthold S.W. Detection of rodent coronaviruses in tissues and cell culture by using polymerase chain reaction. J. Clin. Microbiol. 1991;29:2789–2793. doi: 10.1128/jcm.29.12.2789-2793.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Korner J., Schliephake A., Winter J., Zimprich F., Lassman H., Sedgwick J., Siddell S., Wege H. Nucleocapsid or spike protein-specific CD4+ T lymphocytes protect against coronavirus-induced encephalitis in the absence of CD8+ T cells. J. Immunol. 1991;147:2317–2323. [PubMed] [Google Scholar]
  27. Kunkel F., Herrler G. Structural and functional analysis of the surface protein of human coronavirus OC43. Virology. 1993;195:195–202. doi: 10.1006/viro.1993.1360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lai M.M.C. RNA recombination in animal and plant viruses. Microbiol. Rev. 1992;56:61–79. doi: 10.1128/mr.56.1.61-79.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Luytjes W., Sturman L.S., Breedenbeek P.J., Charite J., van der Zeijst B.A.M., Horzinek M.C., Spaan W.J.M. Primary structure of the glycoprotein E2 of coronavirus MHV-A59 and identification of the trypsin cleavage site. Virology. 1987;161:479–487. doi: 10.1016/0042-6822(87)90142-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mobley L., Evans G., Dailey M.O., Perlman S. Immune response to a murine coronavirus: identification of a homing receptor-negative CD4+ T cell subset that responds to viral glycoproteins. Virology. 1992;187:443–452. doi: 10.1016/0042-6822(92)90446-V. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Parker S.E., Gallagher T.M., Buchmeier M.J. Sequence analysis reveals extensive polymorphism and evidence of deletions within the E2 glycoprotein gene of several strains of murine hepatitis virus. Virology. 1989;173:664–673. doi: 10.1016/0042-6822(89)90579-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Raabe T., Schelle-Prinz B., Siddell S.G. Nucleotide sequence of the gene encoding the spike glycoprotein of human coronavirus HCV 229E. J. Gen. Virol. 1990;71:1065–1073. doi: 10.1099/0022-1317-71-5-1065. [DOI] [PubMed] [Google Scholar]
  33. Rasschaert D., Laude H. The predicted primary structure of the peplomer protein E2 of the porcine coronavirus transmissible gastroenteritis virus. J. Gen. Virol. 1987;68:1883–1890. doi: 10.1099/0022-1317-68-7-1883. [DOI] [PubMed] [Google Scholar]
  34. Robbins S.G., Frana M.F., McGowan J.J., Boyle J.F., Holmes K.V. RNA-binding proteins of coronavirus MHV: detection of monomeric and multimeric N protein with an RNA overlay-protein blot assay. Virology. 1986;150:402–410. doi: 10.1016/0042-6822(86)90305-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schmidt I., Skinner M.A., Siddell S.G. Nucleotide sequence of the gene encoding the surface projection glycoprotein of the coronavirus MHV-JHM. J. Gen. Virol. 1987;68:47–56. doi: 10.1099/0022-1317-68-1-47. [DOI] [PubMed] [Google Scholar]
  36. Spaan W., Cavanaugh D., Horzinek M.C. Coronaviruses: structure and genome expression. J. Gen. Virol. 1988;69:2939–2952. doi: 10.1099/0022-1317-69-12-2939. [DOI] [PubMed] [Google Scholar]
  37. Stauber R., Pfleiderer M., Siddell S. Proteolytic cleavage of the murine coronavirus surface glycoprotein is not required for fusion activity. J. Gen. Virol. 1993;74:183–191. doi: 10.1099/0022-1317-74-2-183. [DOI] [PubMed] [Google Scholar]
  38. Stohlman S.A., Lai M.M.C. Phosphoproteins of murine hepatitis viruses. J. Virol. 1979;32:672–675. doi: 10.1128/jvi.32.2.672-675.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sturman L.S., Holmes K.V., Behnke J. Isolation of coronavirus envelope glycoproteins and interaction with the viral nucleocapsid. J. Virol. 1980;33:449–462. doi: 10.1128/jvi.33.1.449-462.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sturman L.S., Ricard C.S., Holmes K.V. Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: activation of cell-fusing activity of virions by trypsin and separation of two different 90K cleavage fragments. J. Virol. 1985;56:904–911. doi: 10.1128/jvi.56.3.904-911.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Taguchi F. Fusion formation by the uncleaved spike protein of murine coronavirus JHMV variant cl-2. J. Virol. 1993;67:1195–1202. doi: 10.1128/jvi.67.3.1195-1202.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tooze J., Tooze S., Warren G. Replication of coronavirus MHV-A59 in sac-cells: determination of the first site of budding of progeny virions. Eur. J. Cell Biol. 1984;33:281–293. [PubMed] [Google Scholar]
  43. Wang F.-I., Fleming J.O., Lai M.M.C. Sequence analysis of the spike protein gene of murine coronavirus variants: study of genetic sites affecting neuropathogenicity. Virology. 1992;186:742–749. doi: 10.1016/0042-6822(92)90041-M. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wege H., Dorries R. Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes on the peplomer protein (E2) J. Gen. Virol. 1984;65:1931–1942. doi: 10.1099/0022-1317-65-11-1931. [DOI] [PubMed] [Google Scholar]
  45. Wege H., Winter J., Meyermann R. The peplomer protein E2 of coronavirus JHM as a determinant of neurovirulence: definition of critical epitopes by variant analysis. J. Gen. Virol. 1988;69:87–98. doi: 10.1099/0022-1317-69-1-87. [DOI] [PubMed] [Google Scholar]
  46. Williams R.K., Jiang G.-S., Holmes K.V. Vol. 88. 1991. Receptor for mouse hepatitis virus is a member of the carcinoembryonic antigen family of glycoproteins; pp. 5533–5536. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Zhang X., Herbst W., Kousoulas K.G., Storz J. Comparison of the S genes and biological properties of respiratory and enteropathogenic bovine coronaviruses. Arch. Virol. 1994;134:421–426. doi: 10.1007/BF01310579. [DOI] [PMC free article] [PubMed] [Google Scholar]

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