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. 2004 Jan 30;173(2):664–673. doi: 10.1016/0042-6822(89)90579-5

Sequence analysis reveals extensive polymorphism and evidence of deletions within the E2 glycoprotein gene of several strains of murine hepatitis virus

Suezanne E Parker 1, Thomas M Gallagher 1, Michael I Buchmeier 1,1
PMCID: PMC7130524  PMID: 2556846

Abstract

Direct RNA sequence analysis of the E2 gene of wild-type MHV-4 and of neutralization resistant, neuroattenuated variants has identified a polymorphic region with respect to deletions. These variants had large deletions of 142 to 159 amino acids mapping to a localized region in the amino-terminal domain of the peplomer glycoprotein. The nucleotide sequence of the E2 gene for wild-type strain MHV-4 was found to be very similar to that of MHV-JHM but had an insertion of 423 nucleotides resulting in the addition of a stretch of 141 unique amino acids in the amino-terminal domain of E2. We propose that deletions reflect a major source of heterogeneity in the E2 protein of MHV.

References

  1. Bark R.S., Chien C.-K., Stohlman S.A., Lai M.M.C. Analysis of intracellular small RNAs of mouse hepatitis virus: Evidence for discontinuous transcription. Virology. 1987;156:342–354. doi: 10.1016/0042-6822(87)90414-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cavanagh D. Coronavirus IBV: Structural characterization of the spike protein. J. Gen. Virol. 1983;64:2577–2583. doi: 10.1099/0022-1317-64-12-2577. [DOI] [PubMed] [Google Scholar]
  3. Chirgwin J.M., Przyblya A.E., MacDonald R.J., Rutter W.J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979;18:5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Devereux J., Haberli P., Smithies 0. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984;12:387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubois-Dalcq M.E., Doller E.W., Haspel M.V., Holmes K.W. Cell tropism and expression of mouse hepatitis virus (MHV) in mouse spinal cord cultures. Virology. 1982;119:317–331. doi: 10.1016/0042-6822(82)90092-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fazakerley J.F., Ross A.M. Computer analysis suggests a role for signal sequences in processing polyproteins of enveloped RNA viruses and as a mechanism of viral fusion. Virus Genes. 1989;2:219–235. doi: 10.1007/BF00125340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fleming J.O., Stohlman S.A., Harmon R.C., Lai M.M.C., Frelinger J.A., Weiner L.P. Antigenic relationships of murine coronaviruses: Analysis using monoclonal antibodies to JHM (MHV-4) virus. Virology. 1983;131:296–307. doi: 10.1016/0042-6822(83)90498-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fleming J.O., Trousdale M.D., El-Zaatari F.A.K., Stohlman S.A., Weiner L.P. 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]
  10. Hamlyn P.H., Gait M.J., Milstein C. Complete sequence of an immunoglobulin mRNA using specific priming and the dideoxynucleotide method of RNA sequencing. Nucleic Acids Res. 1981;9:4485–4494. doi: 10.1093/nar/9.18.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Haspel M.V., Lampert P.W., Oldstone M.B.A. Vol. 75. 1978. Temperature sensitive mutants of mouse hepatitis virus produce a high incidence of demyelination; pp. 4033–4036. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Keck J.G., Matsushima G.K., Making S., Fleming J.O., Vannier D.M., Stohlman S.A., Lai M.M.C. In vivo RNA-RNA recombination of coronavirus in mouse brain. J. Virol. 1988;62:1810–1813. doi: 10.1128/jvi.62.5.1810-1813.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knobler R.L., Tunison L.A., Lampert P.W., Oldstone M.B.A. Selected mutants of mouse hepatitis virus type 4 (JHM strain) induce different CNS diseases. Amer. J. Pathol. 1982;109:157–168. [PMC free article] [PubMed] [Google Scholar]
  14. Kyte J., Doolittle R.F. A simple method for displaying the hydropathic character of a protein. J. Mol. Biol. 1982;157:105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  15. Lai M.M.C., Baric R.S., Making S., Keck J.G., Egbert J., Leibowitz J.L., Stohlman S.A. Recombination between nonsegmented RNA genomes of murine coronaviruses. J. Virol. 1985;56:449–456. doi: 10.1128/jvi.56.2.449-456.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lampert P.W., Sims J.K., Kniazeff A.J. Mechanism of demyelination in JHM virus encephalomyelitis: Electron microscopic studies. Acta Neuropathol. 1973;24:76–85. doi: 10.1007/BF00691421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Luytjes W., Sturman L.S., Bredenbeek 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]
  18. Luytjes W., Bredenbeek P.J., Noten A.F.H., Horzinek M.C., Spaan W.J.M. Sequence of mouse hepatitis virus A59 mRNA2: Indications for RNA recombination between coronaviruses and influenza C virus. Virology. 1988;166:415–422. doi: 10.1016/0042-6822(88)90512-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Makino S., Taguchi F., Fujiwara K. Defective interfering particles of mouse hepatitis virus. Virology. 1984;133:9–17. doi: 10.1016/0042-6822(84)90420-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Makino S., Keck J.G., Stohlman S.A., Lai M.M.C. High frequency of RNA recombination of murine coronaviruses. J. Virol. 1986;57:729–737. doi: 10.1128/jvi.57.3.729-737.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Makino S., Fleming J.O., Keck J.G., Stohlman S.A., Lai M.M.C. Vol. 84. 1987. RNA recombination of coronaviruses: Localization of neutralizing epitopes and neuropathogenic determinants on the carboxy terminus of peplomers; pp. 6567–6571. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morris V.L., Tieszer C., Mackinnon J., Percy D. Characterization of coronavirus JHM variants isolated from Wistar Furth rats with a viral induced demyelinating disease. Virology. 1989;169:127–136. doi: 10.1016/0042-6822(89)90048-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Neuberger A., Gottschalk A., Marshall R.O., Spiro R.G. Carbohydrate-peptide linkages in glycoproteins and methods for their elucidation. In: Gottschalk A., editor. The Glycoproteins: Their Composition, Structure and Function. Elsevier; Amsterdam: 1972. pp. 450–490. [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A.R. Vol. 74. 1977. DNA sequencing with chain terminating inhibitors; pp. 5463–5467. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schmidt I., Skinner M., Siddel S. Nucleotide sequence of the gene encoding the surface projection glycoprotein of coronavirus MHV-JHM. J. Gen. Virol. 1987;68:47–56. doi: 10.1099/0022-1317-68-1-47. [DOI] [PubMed] [Google Scholar]
  26. Siddel S., Wege H., ter Meulen V. The structure and replication of coronaviruses. Curr. Topics Microbiol. Immunol. 1982;99:131–163. doi: 10.1007/978-3-642-68528-6_4. [DOI] [PubMed] [Google Scholar]
  27. Skinner M.A., Ebner D., Siddel S.G. Coronavirus MHV-JHM mRNA 5 has a sequence arrangement which potentially allows a translation of a second downstream open reading frame. J. Gen. Virol. 1985;66:581–592. doi: 10.1099/0022-1317-66-3-581. [DOI] [PubMed] [Google Scholar]
  28. Skinner M.A., Siddel S.G. Coding sequence of coronavirus MHV-JHM mRNA 4. J. Gen. Virol. 1985;66:593–596. doi: 10.1099/0022-1317-66-3-593. [DOI] [PubMed] [Google Scholar]
  29. Stohlman S.A., Brayton P.R., Fleming J.O., Weiner L.P., Lai M.M.C. Murine coronaviruses: Isolation and characterization of two plaque morphology variants of the JHM neurotropic strain. J. Gen. Virol. 1982;63:265–275. doi: 10.1099/0022-1317-63-2-265. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Taguchi F., Siddel S.G., Wege H., ter Meulen V. Characterization of a variant virus selected in rat brains after infection by coronavirus mouse hepatitis virus JHM. J. Virol. 1985;54:429–435. doi: 10.1128/jvi.54.2.429-435.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Taguchi F., Massa P.T., ter Meulen V.C. Characterization of a variant virus isolated from neural cell culture after infection of mouse coronavirus JHMV. Virology. 1986;155:267–270. doi: 10.1016/0042-6822(86)90187-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Taguchi F., Fleming J.O. Comparison of six different murine coronavirus JHM variants by monoclonal antibodies against the E2 glycoprotein. Virology. 1989;169:233–235. doi: 10.1016/0042-6822(89)90061-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Talbot P.J., Buchmeier M.J. Antigenic variation among murine coronaviruses: Evidence for polymorphism on the peplomer glycoprotein, E2. Virus Res. 1985;2:317–328. doi: 10.1016/0168-1702(85)90028-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986;14:4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wege H., Siddel S., ter Meulen V. The biology and pathogenesis of coronaviruses. Curr. Topics Microbiol. Immunol. 1982;99:165–200. doi: 10.1007/978-3-642-68528-6_5. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Weiner L.P. Pathogenesis of demyelination induced by a mouse hepatitis virus (JHM virus) Arch. Neurol. 1973;28:298–303. doi: 10.1001/archneur.1973.00490230034003. [DOI] [PubMed] [Google Scholar]

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