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. 2002 May 25;294(1):222–236. doi: 10.1006/viro.2001.1307

The Spike but Not the Hemagglutinin/Esterase Protein of Bovine Coronavirus Is Necessary and Sufficient for Viral Infection

Rada Popova 1, Xuming Zhang 1,1
PMCID: PMC7131450  PMID: 11886280

Abstract

The spike (S) and hemagglutinin/esterase (HE) of bovine coronavirus (BCV) are the two envelope proteins that recognize the same receptor-determinant of 9-O-acetylneuraminic acid on host cells. However, the precise and relative roles of the two proteins in BCV infectivity remain elusive. To unequivocally determine their roles in viral cytopathogenicity, we developed a system in which phenotypically chimeric viruses were generated by infecting a closely related mouse hepatitis virus (MHV) in cells that stably express an individual BCV protein (S or HE). The chimeric viruses were then used to infect human rectal tumor (HRT)-18 cells that are permissive to BCV but are nonsusceptible to MHV. Using this approach, we found that the chimeric virus containing the BCV S protein on the virion surface entered and replicated in HRT-18 cells; this was specifically blocked by prior treatment of the virus with a neutralizing antibody specific to the BCV S protein, indicating that the BCV S protein is responsible for initiating chimeric virus infection. In contrast, chimeric viruses that contain biologically active and functional BCV HE protein on the surface failed to enter HRT-18 cells, indicating that the BCV HE protein alone is not sufficient for BCV infection. Taken together, these results demonstrate that the S protein but not the HE protein of BCV is necessary and sufficient for infection of the chimeric viruses in HRT-18 cells, suggesting that BCV likely uses the S protein as a primary vehicle to infect permissive cells.

References

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.Boireau P., Cruciere 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]
  • 4.Cavanagh D. The coronavirus surface glycoprotein. In: Siddell S.G., editor. The Coronaviridae. Plenum; New York: 1995. pp. 73–113. [Google Scholar]
  • 5.Deregt D., Babiuk L.A. Monoclonal antibodies to bovine coronavirus: Characteristics and topographical mapping of neutralizing epitopes on the E2 and E3 glycoproteins. Virology. 1987;161:410–420. doi: 10.1016/0042-6822(87)90134-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Deregt D., Gifford G.A., Ijaz M.K., Watts T.C., Gilchrist J.E., Haines D.M., Babiuk L.A. Monoclonal antibodies to bovine coronavirus glycoproteins E2 and E3: Demonstration of in vivo virus-neutralizing activity. J. Gen. Virol. 1989;70:993–998. doi: 10.1099/0022-1317-70-4-993. [DOI] [PubMed] [Google Scholar]
  • 7.Gagneten S., Gout O., Dubois-Dalco M., Rottier P., Rossen J., Holmes K.V. Interaction of mouse hepatitis virus (MHV) spike glycoprotein with receptor glycoprotein MHVR is required for infection with an MHV strain that expresses the hemagglutinin-esterase glycoprotein. J. Virol. 1995;69:889–895. doi: 10.1128/jvi.69.2.889-895.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Godeke G.-J., de Haan C.A.M., Rossen J.W.A., Vennema H., Rottier P.J.M. Assembly of spikes into coronavirus particles is mediated by the carboxy-terminal domain of the spike protein. J. Virol. 2000;74:1566–1571. doi: 10.1128/jvi.74.3.1566-1571.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Herrler G., Rott R., Klenk H.D., Muller H.P., Shukda A.K., Schauer R. The receptor-destroying enzyme of influenza C is virus neuraminidate-O-acetyl-esterase. EMBO J. 1985;4:1503–1506. doi: 10.1002/j.1460-2075.1985.tb03809.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Herrler G., Durkop I., Becht H., Klenk H.D. The glycoprotein of influenza C virus is the haemagglutinin, esterase and fusion factor. J. Gen. Virol. 1988;69:839–846. doi: 10.1099/0022-1317-69-4-839. [DOI] [PubMed] [Google Scholar]
  • 11.Herrler G., Klenk H.D. Structure and function of the HEF glycoprotein of influenza C virus. Adv. Virus Res. 1991;40:213–234. doi: 10.1016/S0065-3527(08)60280-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Hirano N., Fujiwara K., Hino S., Matsumoto M. Replication and plaque formation of mouse hepatitis virus (MHV-2) in mouse cell line DBT culture. Arch. Gesamte Virusforsch. 1974;44:298–302. doi: 10.1007/BF01240618. [DOI] [PubMed] [Google Scholar]
  • 13.Hussain K.A., Storz J., Kousoulas K.G. Comparison of bovine coronavirus (BCV) antigens: Monoclonal antibodies to the spike glycoprotein distinguish between vaccine and wild-type strains. Virology. 1991;183:225–442. doi: 10.1016/0042-6822(91)90163-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.King B., Potts B.J., Brian D.A. Bovine coronavirus hemagglutinin protein. Virus Res. 1985;2:53–59. doi: 10.1016/0168-1702(85)90059-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Kuo L., Godeke G.J., Raamsman M.J., Masters P.S., Rottier P.J. Retargeting of coronavirus by substitution of the spike glycoprotein ectodomain: Crossing the host cell species barrier. J. Virol. 2000;74:1393–1406. doi: 10.1128/jvi.74.3.1393-1406.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Lai M.M.C., Cavanagh D. The molecular biology of coronaviruses. Adv. Virus Res. 1997;48:1–100. doi: 10.1016/S0065-3527(08)60286-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Liao C.L., Lai M.M.C. Requirement of the 5′-end genomic sequence as an upstream cis-acting element for coronavirus subgenomic mRNA transcription. J. Virol. 1994;68:4727–4737. doi: 10.1128/jvi.68.8.4727-4737.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Liao C.-L., Zhang X.M., Lai M.M.C. Coronavirus defective-interfering RNA as an expression vector: The generation of a pseudorecombinant mouse hepatitis virus expressing hemagglutinin-esterase. Virology. 1995;208:319–327. doi: 10.1006/viro.1995.1155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Luytjes W., Bredenbeek P.J., Noten A.F.H., Horzinek M.C., Spaan W.J.M. Sequence of mouse hepatitis virus A59 mRNA 2: Indications for RNA-recombination between coronavirus and influenza C virus. Virology. 1988;166:415–422. doi: 10.1016/0042-6822(88)90512-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Parker M.D., Yoo D., Cox G.J., Babiuk L.A. Primary structure of the S peplomer gene of bovine coronavirus and surface expression in insect cells. J. Gen. Virol. 1990;71:263–270. doi: 10.1099/0022-1317-71-2-263. [DOI] [PubMed] [Google Scholar]
  • 21.Rottier P., Branderberg D., Armstrong J., van der Zeijst B., Warren G. Assembly in vitro of a spanning membrane protein of the endoplasmic reticulum: The E1 glycoprotein of coronavirus mouse hepatitis virus A59. Proc. Natl. Acad. Sci. USA. 1984;81:1421–1425. doi: 10.1073/pnas.81.5.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Sanchez C.M., Izeta A., Sanchez-Morgado J.M., Alonso S., Sola I., Balasch M., Plana-Duran J., Enjuanes L. Targeted recombination demonstrates that the spike gene of transmissible gastroenteritis coronavirus is a determinant of its enteric tropism and virulence. J. Virol. 1999;73:7607–7618. doi: 10.1128/jvi.73.9.7607-7618.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Schultze B., Gross H.J., Brossmer R., Klenk H.D., Herrler G. Hemagglutinating encephalomyelitis virus attaches to N-acetyl-9-O-acetylneuraminic acid-containing receptors on erythrocytes: Comparison with bovine coronavirus and influenza C virus. Virus Res. 1990;16:185–194. doi: 10.1016/0168-1702(90)90022-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Schultze B., Gross H.J., Brossmer R., Herrler G. The S protein of bovine coronavirus is a hemagglutinin recognizing 9-O-acetylated sialic acid as a receptor determinant. J. Virol. 1991;65:6232–6237. doi: 10.1128/jvi.65.11.6232-6237.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Schultze B., Herrler G. Bovine coronavirus uses N-acetyl-9-O-acetylneuraminic acid as a receptor determinant to initiate the infection of cultured cells. J. Gen. Virol. 1992;73:901–906. doi: 10.1099/0022-1317-73-4-901. [DOI] [PubMed] [Google Scholar]
  • 26.Schultze B., Herrler G. Recognition of cellular receptors by bovine coronavirus. Arch. Virol. Suppl. 1994;9:451–459. doi: 10.1007/978-3-7091-9326-6_44. [DOI] [PubMed] [Google Scholar]
  • 27.Shieh C.K., Lee H.J., Yokomori K., La Monica N., Makino S., Lai M.M.C. Identification of a new transcriptional initiation site and the corresponding functional gene 2b in the murine coronavirus RNA genome. J. Virol. 1989;63:3729–3736. doi: 10.1128/jvi.63.9.3729-3736.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Spaan W.J.M., Rottier P.J.M., Horzinek M.C., van der Zeijst B.A.M. Isolation and identification of virus-specific mRNA in cells infected with mouse hepatitis virus (MHV-A59) Virology. 1981;108:424–434. doi: 10.1016/0042-6822(81)90449-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.St. Cyr-Coats K., Storz J. Bovine coronavirus induced cytopathic expression and plaque formation: Host cell and virus strain determine trypsin dependence. J. Vet. Med. 1988;35:48–56. doi: 10.1111/j.1439-0450.1988.tb00465.x. [DOI] [PubMed] [Google Scholar]
  • 30.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]
  • 31.Storz J., Rott R., Kaluza G. Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment. Infect. Immun. 1981;31:1214–1222. doi: 10.1128/iai.31.3.1214-1222.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Storz J., Zhang X.M., Rott R. Comparison of hemagglutination, receptor-destroying, and acetylesterase activities of avirulent and virulent bovine coronavirus strains. Arch. Virol. 1992;125:193–204. doi: 10.1007/BF01309637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Sturman L.S., Holmes K.V., Behnke J. Isolation of coronavirus envelop 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]
  • 34.Vennema H., Godeke G.J., Rossen J.W.A., Voorhout W.F., Horzinek M.C., Opstelten D.J.E., Rottier P.J.M. Nucleocapsid-independent assembly of coronavirus-like particles by co-expression of viral envelope protein genes. EMBO J. 1996;15:2020–2028. doi: 10.1002/j.1460-2075.1996.tb00553.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Vlasak R., Luytjes W., Leider J., Spaan W., Palese P. The E3 protein of bovine coronavirus is a receptor-destroying enzyme with acetylesterase activity. J. Virol. 1988;62:4686–4690. doi: 10.1128/jvi.62.12.4686-4690.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Wang Y., Zhang X.M. The nucleocapsid protein of coronavirus mouse hepatitis virus interacts with the cellular heterogeneous nuclear ribonucleoprotein A1 in vitro and in vivo. Virology. 1999;265:96–109. doi: 10.1006/viro.1999.0025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Yokomori K., Monica N.L., Makino S., Shieh C.K., Lai M.M.C. Biosynthesis, structure, and biological activities of envelope protein gp65 of murine coronavirus. Virology. 1989;173:683–691. doi: 10.1016/0042-6822(89)90581-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 38.Yokomori K., Banner L.R., Lai M.M.C. Heterogeneity of gene expression of hemagglutinin-esterase (HE) protein of murine coronaviruses. Virology. 1991;183:647–657. doi: 10.1016/0042-6822(91)90994-M. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Yoo D.W., Parker M.D., Babiuk L.A. The S2 subunit of the spike glycoprotein of bovine coronavirus mediates membrane fusion in insect cells. Virology. 1991;180:395–399. doi: 10.1016/0042-6822(91)90045-D. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Yu X., Bi W., Weiss S.R., Leibowitz J.L. Mouse hepatitis virus gene 5b protein is a new virion envelope protein. Virology. 1994;202:1018–1023. doi: 10.1006/viro.1994.1430. [DOI] [PubMed] [Google Scholar]
  • 41.Zhang X.M., Kousoulas K.G., Storz J. The hemagglutinin/esterase glycoprotein of bovine coronaviruses: Sequence and functional comparisons between virulent and avirulent strains. Virology. 1991;185:847–852. doi: 10.1016/0042-6822(91)90557-R. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Zhang X.M., Kousoulas K.G., Storz J. Comparison of the nucleotide and deduced amino acid sequences of the S genes specified by virulent and avirulent strains of bovine coronaviruses. Virology. 1991;183:397–404. doi: 10.1016/0042-6822(91)90154-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Zhang X.M., Herbst W., Kousoulas K.G., Storz J. Biological and genetic characterization of a hemagglutinating coronavirus isolated from a diarrhoeic child. J. Med. Virol. 1994;44:152–161. doi: 10.1002/jmv.1890440207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Zhang X.M., Hinton D.R., Cue D., Stohlman S.A., Lai M.M.C. Expression of gamma interferon by a coronavirus defective-interfering RNA vector and its effect on viral replication, spread and pathogenicity. Virology. 1997;233:327–338. doi: 10.1006/viro.1997.8598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Zhang X.M., Hinton D.R., Parra B., Park S., Liao C.L., Lai M.M.C., Stohlman S.A. Expression of hemagglutinin/esterase by a mouse hepatitis virus coronavirus defective-interfering RNA alters viral pathogenesis. Virology. 1998;242:170–183. doi: 10.1006/viro.1997.8993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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