Skip to main content
PMC home page
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 1994;135(3):319–331. doi: 10.1007/BF01310017

Comparative sequence analysis of a polymorphic region of the spike glycoprotein S1 subunit of enteric bovine coronavirus isolates

M R Rekik 1, S Dea 1
PMCID: PMC7086735  PMID: 7979970

Summary

Complementary oligonucleotide primers which flank a 1146-nucleotide gene fragment (S1B: nt 1185 to 2333) encompassing a polymorphic region (nt 1368 to 1776) of the S1 subunit of bovine coronavirus spike glycoprotein were used for enzymatic amplification by PCR. We chose four clinical isolates, recovered from cases of epidemic diarrhea in neonatal calves in Québec dairy herds between 1987–1990, to specifically amplify and analyze their sequences in the selected genomic area. Nucleotide sequence analysis of the four clinical isolates indicated that their S1B gene fragments were highly conserved. We also compared the S1B gene sequences of the Québec BCV isolates to the published corresponding sequences from BCV-L9 [37], BCV-MEB [1], and BCV-F15 [3] reference strains. A high degree of similarity was demonstrated for all viruses, no deletions or insertions were observed, and the only variations that were identified consisted of nucleotide substitutions. The differing nucleotides and amino acids (aa) were not distributed randomly over the entire sequence but rather were clustered in the polymorphic region. Of these, four sporadic aa changes were located in antigenic domain II (aa residues 517 to 720) of S1. This correlates with varied antigenicity observed among the BCV Québec isolates when reacting with MAbs directed against the S glycoprotein of the Mebus strain. The other mutations seem to be fixed in all Québec isolates.

Keywords: Nucleotide Sequence Analysis, Dairy Herd, Comparative Sequence Analysis, Polymorphic Region, Complementary Oligonucleotide

References

  • 1.Abraham S, Kienzle TE, Lapps W, Brian DA. Deduced sequence of the bovine coronavirus spike protein and identification of the internal proteolytic cleavage site. Virology. 1991;176:296–301. doi: 10.1016/0042-6822(90)90257-R. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Benfield DA, Saif L. Cell culture propagation of a coronavirus isolated from cows with winter dysentery. J Clin Microbiol. 1990;28:1454–1457. doi: 10.1128/jcm.28.6.1454-1457.1990. [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.Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform. Anal Biochem. 1987;162:156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  • 5.Cyr-Coats K, Storz J, Hussain KA, Schnorr KL. Structural proteins of bovine coronavirus strain L9: Effects of the host cell and trypsin treatment. Arch Virol. 1988;103:35–45. doi: 10.1007/BF01319807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Dea S, Verbeek AJ, Tijssen P. Antigenic and genomic relationships among turkey and bovine enteric coronaviruses. J Virol. 1990;64:3112–3118. doi: 10.1128/jvi.64.6.3112-3118.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Dea S, Roy RS, Begin ME. Bovine coronavirus isolation and cultivation in continuous cell lines. Am J Vet Res. 1980;41:30–38. [PubMed] [Google Scholar]
  • 8.Dea S, Garzon S, Tijssen P. Isolation and trypsin-enhanced propagation of turkey enteric (bluecomb) coronaviruses in a continuous human rectal adenocarcinoma cell line. Am J Vet Rec. 1989;50:1310–1318. [PubMed] [Google Scholar]
  • 9.Deregt D, Babiuk LA. 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]
  • 10.Deregt D, Parker MD, Cox GC, Babiuk LA. Mapping of neutralizing epitopes to fragments of the bovine coronavirus E2 protein by proteolysis of antigen-antibody complexes. J Gen Virol. 1989;70:647–658. doi: 10.1099/0022-1317-70-3-647. [DOI] [PubMed] [Google Scholar]
  • 11.Deregt D, Sabara M, Babiuk LA. Structural proteins of bovine coronavirus and their intracellular processing. J Gen Virol. 1987;68:2863–2877. doi: 10.1099/0022-1317-68-11-2863. [DOI] [PubMed] [Google Scholar]
  • 12.Gallagher TM, Parker SE, Buchmeier MJ. Neutralization-resistant variants of aneurotropic 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]
  • 13.Hogue BG, King B, Brian DA. Antigenic relationships among proteins of bovine coronavirus, human respiratory coronavirus OC43, and mouse hepatitis coronavirus A59. J Virol. 1984;51:384–388. doi: 10.1128/jvi.51.2.384-388.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Hsu M-C, Scheid A, Choppin PW. Protease activation mutants of Sendai viruses: sequence analysis of the mRNA of the fusion protein (F) gene and direct identification of the cleavage-activation site. Virology. 1987;156:84–90. doi: 10.1016/0042-6822(87)90438-7. [DOI] [PubMed] [Google Scholar]
  • 15.Hussain KA, Storz J, Kousoulas KG. Comparison of bovine coronavirus (BCV) antigens: monoclonal antibodies to the spike protein distinguish between vaccine and wild-type strains. Virology. 1991;183:442–445. doi: 10.1016/0042-6822(91)90163-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.King B, Potts BJ, Brian DA. 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]
  • 17.Laporte J, Bobulesco P, Rossi F. Une lignée particulièrement sensible à la réplication du coronavirus entéritique bovin: les cellules HRT-18. CR Acad Sci (III) Paris. 1980;290D:623–626. [PubMed] [Google Scholar]
  • 18.Mebus CA, Stair EL, Rhodes MB, Twiehaus MJ. Neonatal calf diarrhoea: propagation, attenuation, and characteristics of a coronavirus-like agent. Am J Vet Res. 1973;34:145–150. [PubMed] [Google Scholar]
  • 19.Michaud L, Dea S. Characterization of monoclonal antibodies to bovine enteric coronavirus and antigenic variability among Quebec isolates. Arch Virol. 1993;131:455–465. doi: 10.1007/BF01378646. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Parker MD, Yoo D, Cox GJ, Babiuk LA. 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.Parker SE, Gallagher TM, Buchmeier MJ. Sequence analysis reveals extensive polymorphism and evidence of deletion within 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]
  • 22.Reynolds DJ, Debney TG, Hall GA, Thomas LH, Parsons KR. Studies on the relationship between coronaviruses from the intestinal and respiratory tracts of calves. Arch Virol. 1985;85:71–83. doi: 10.1007/BF01317007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Rott R, Orlich M, Klenk H-D, Wang ML, Skehel JJ, Wiley DC. Studies on the adaptation of influenza viruses to MDCK cells. EMBO J. 1984;3:3329–3332. doi: 10.1002/j.1460-2075.1984.tb02299.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Saif LJ, Redman DR, Moorhead PD, Theil KW. Experimentally-induced coronavirus infections in calves: viral replication in the respiratory and intestinal tracts. Am J Vet Res. 1986;47:1426–1432. [PubMed] [Google Scholar]
  • 25.Saif LJ, Brock KV, Redman DR, Kohler EM. Winter dysentery in dairy herds: electron microscopic and serological evidence for an association with coronavirus infection. Vet Rec. 1991;128:447–449. doi: 10.1136/vr.128.19.447. [DOI] [PubMed] [Google Scholar]
  • 26.Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA. 1977;74:5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Sharpee RL, Mebus CA, Bass EP. Characterization of a calf diarrheal coronavirus. Am J Vet Res. 1976;37:1031–1041. [PubMed] [Google Scholar]
  • 28.Siddell S, Wege H, Ter Meulen V. The biology of coronaviruses. J Gen Virol. 1983;64:761–776. doi: 10.1099/0022-1317-64-4-761. [DOI] [PubMed] [Google Scholar]
  • 29.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]
  • 30.Sturman LS, Ricard CS, Holmes KV. 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]
  • 31.Sturman LS, Holmes KV. The molecular biology of coronaviruses. Adv Virus Res. 1983;28:35–112. doi: 10.1016/S0065-3527(08)60721-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Tompkins WAF, Watrach AW, Schmale JD, Schultz RM, Harris JA. Cultural and antigenic properties of newly established cell strains derived from adenocarcinomas of the human colon and rectum. J Natl Cancer Inst. 1974;52:101–106. doi: 10.1093/jnci/52.4.1101. [DOI] [PubMed] [Google Scholar]
  • 33.Vautherot JF, Laporte J, Madelaine MF, Bobulesco P, Roseto A. Antigenic and polypeptide structure of bovine enteric coronavirus as defined by monoconal antibodies. Adv Exp Med Biol. 1984;173:117–132. doi: 10.1007/978-1-4615-9373-7_11. [DOI] [PubMed] [Google Scholar]
  • 34.Vautherot JF, Madelaine MF, Boireau P, Laporte J. Bovine coronavirus peplomer glycoproteins: detailed antigenic analysis of S1, S2 and HE. J Gen Virol. 1992;73:1725–1737. doi: 10.1099/0022-1317-73-7-1725. [DOI] [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.Yoo D, Parker MD, Song J, Cox GJ, Deregt D, Babiuk LA. Structural analysis of the conformational domains involved in neutralization of bovine coronavirus using deletion mutants of the spike glycoprotein S1 subunit expressed by recombinant baculoviruses. Virology. 1991;183:91–98. doi: 10.1016/0042-6822(91)90121-Q. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Zhang X, Kousoulas KG, 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]

Articles from Archives of Virology are provided here courtesy of Nature Publishing Group

RESOURCES