Skip to main content
Springer Nature - PMC COVID-19 Collection logoLink to Springer Nature - PMC COVID-19 Collection
. 1979;59(1):47–57. doi: 10.1007/BF01317894

Structural polypeptides of the enteropathogenic bovine coronavirus strain LY-138

I Hajer 1, J Storz 1,3
PMCID: PMC7086941  PMID: 218536

Summary

The bovine coronavirus strain LY-138 was purified by differential as well as velocity and isopycnic centrifugation in sucrose or CsCl gradients. The substrate for purification was contents of the small intestine of experimentally inoculated calves. This strain is highly enteropathogenic, but it could not yet be propagated in cultured cells. Intact virions had a density of 1.245 g/cm3 in CsCl and 1.185 g/cm3 in sucrose. A spherical core-like structure with an average diameter of 82 nm remaining after treatment with chloroform had a density of 1.299 g/cm3 in CsCl and 1.201 g/cm3 in sucrose.

Seven distinct bands of polypeptides and 4 shoulders were detected after electrophoresis of SDS-solubilized virions in polyacrylamide gels. The approximate molecular weights ranged from 110,000 to 36,000. Four of the bands gave a PAS positive reaction. These 4 glycoproteins and an additional protein with an approximate molecular weight of 70,000 were removed by chloroform treatment. The remaining core-like structure contained the 2 polypeptides VP3 and VP7.

Keywords: Sucrose, Chloroform, Polypeptide, Small Intestine, Polyacrylamide

Footnotes

With 5 Figures

References

  • 1.Berry D. M., Cruickshank J. G., Chu H. P., Wells R. J. H. The structure of infectious bronchitis virus. Virology. 1964;23:403–407. doi: 10.1016/0042-6822(64)90263-6. [DOI] [PubMed] [Google Scholar]
  • 2.Binn, L. N., Lazar, E. C., Keenan, K. P., Huxsoll, D. L., Marchwicki, R. H., Strano, A. J.: Recovery and characterization of a coronavirus from military dogs with diarrhea. In: Animal Health (Proc. of the 78th Annual Meeting, U.S. Anim. Health Assoc., Roanoke, Virginia, 1974), 359–366 (1975). [PubMed]
  • 3.Bolognesi D. P., Montelaro R. C., Frank H., Schäfer W. Assembly of type C Oncornaviruses: A model. Science. 1978;199:183–186. doi: 10.1126/science.202022. [DOI] [PubMed] [Google Scholar]
  • 4.Bolognesi D. P., Gelderblom H., Bauer H., Mölling K., Huper G. Polypeptides of avian RNA tumor viruses. V. Analysis of the virus cores. Virology. 1972;47:567–576. doi: 10.1016/0042-6822(72)90546-6. [DOI] [PubMed] [Google Scholar]
  • 5.Caliguiri L. A., Klenk H. D., Choppin P. W. The proteins of the parainfluenza virus SV 5. I. Separation of virion polypeptides by polyacrylamide gel electrophoresis. Virology. 1969;39:460–466. doi: 10.1016/0042-6822(69)90094-4. [DOI] [PubMed] [Google Scholar]
  • 6.Caul E. O., Paver W. K., Clarke S. K. R. Coronavirus particles in faeces from patients with gastroenteritis. Lancet. 1975;i:1192–1193. doi: 10.1016/S0140-6736(75)93176-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Coffin J. M., Temin H. M. Comparison of Rous sarcoma virus specific deoxy-ribonucleic acid polymerases in virions of Rous sarcoma virus and in Rous sarcoma virus infected chicken cells. J. Virol. 1971;7:625–634. doi: 10.1128/jvi.7.5.625-634.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Doughri A. M., Storz J. Light and ultrastructural pathologic changes in intestinal coronavirus infection of newborn calves. Zentralbl. Vet. Med. (B) 1977;24:367–385. doi: 10.1111/j.1439-0450.1977.tb01011.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Doughri A. M., Storz J., Hajer I., Fernando H. S. Morphology and morphogenesis of a coronavirus infecting intestinal epithelial cells of newborn calves. Expt. mol. Pathol. 1976;25:355–370. doi: 10.1016/0014-4800(76)90045-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Doyle L. P., Hutchings L. M. A transmissible gastro-enteritis in pigs. J. Amer. vet. Med. Ass. 1946;108:257–259. [PubMed] [Google Scholar]
  • 11.Garwes D. J., Pocock D. H., Pike B. W. Isolation of subviral components from transmissible gastro-enteritis virus. J. gen. Virol. 1976;32:283–294. doi: 10.1099/0022-1317-32-2-283. [DOI] [PubMed] [Google Scholar]
  • 12.Garwes D. J., Pocock D. H. The polypeptide structure of transmissible gastroenteritis virus. J. gen. Virol. 1975;29:25–34. doi: 10.1099/0022-1317-29-1-25. [DOI] [PubMed] [Google Scholar]
  • 13.Garwes D. J., Pocock D. H., Wijaszka T. M. Identification of heat-dissociable RNA complexes in two porcine coronaviruses. Nature. 1975;257:508–510. doi: 10.1038/257508a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Hajer I., Storz J. Antigens of bovine coronavirus strain LY-138 and their diagnostic properties. Amer. J. vet. Res. 1978;39:441–444. [PubMed] [Google Scholar]
  • 15.Hierholzer J. C. Purification and biophysical properties of human coronavirus 229 E. Virology. 1976;75:155–165. doi: 10.1016/0042-6822(76)90014-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Hierholzer J. C., Palmer E. L., Whitfield S. G., Kaye H. S., Dowdle W. R. Protein composition of coronavirus OC43. Virology. 1972;48:516–527. doi: 10.1016/0042-6822(72)90062-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Kaye H. S., Hierholzer J. C., Dowdle W. R. Purification and further characterization of an “IBV-like” virus (coronavirus) Exp. Biol. Med. 1970;135:457–463. doi: 10.3181/00379727-135-35074. [DOI] [PubMed] [Google Scholar]
  • 18.Kennedy D. A., Johnson-Lussenburg M. Isolation and morphology of the internal component of human coronavirus strain 229 E. Intervirology. 1976;6:197–206. doi: 10.1159/000149474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Lewis L. D., Phillips R. W. Diarrheic induced changes in intracellular and extracellular ion concentrations in neonatal calves. Ann. Rech. Vétér. 1973;4:99–111. [Google Scholar]
  • 20.Lomniczi B., Kennedy I. Genome of infectious bronchitis virus. J. Virol. 1977;24:99–107. doi: 10.1128/jvi.24.1.99-107.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. Protein measurement with the follin phenol reagent. J. biol. Chem. 1951;193:265–275. [PubMed] [Google Scholar]
  • 22.Luna L. G. Manual of Histopathologic staining methods of the Armed Forces Institute of Pathology. 3rd Ed. New York: McGraw-Hill Book Co.; 1968. [Google Scholar]
  • 23.MacNaughton M. R., Madge M. H., Davies H. A., Dourmashkin R. R. Polypeptides of the surface projections and the ribonucleoprotein of avian infectious bronchitis virus. J. Virol. 1977;24:821–825. doi: 10.1128/jvi.24.3.821-825.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Maizel J. V., Summers D. F., Scharff M. D. SDS-acrylamide gel electrophoresis and its application to the proteins of poliovirus and adenovirus-infected human cells. J. Cell Physiol. 1970;76:273–288. doi: 10.1002/jcp.1040760307. [DOI] [PubMed] [Google Scholar]
  • 25.McIntosh K. Coronaviruses: A comparative review. Curr. Top. Microbiol. Immun. 1974;63:84–129. [Google Scholar]
  • 26.Mebus C. A., Stair E. L., Rhodes M. B., Twiehaus M. J. Pathology of neonatal calf diarrhea induced by a coronavirus-like agent. Vet. Pathol. 1973;10:45–64. doi: 10.1177/030098587301000105. [DOI] [PubMed] [Google Scholar]
  • 27.Phillips R. W., Lewis L. D. Viral induced changes in intestinal transport and resultant body fluid alterations in neonatal calves. Ann. Rech. Vétér. 1973;4:87–98. [Google Scholar]
  • 28.Rowe W. P., Hartley J. W., Capps W. I. Mouse hepatitis virus infection as a highly contagious, prevalent, enteric infection of mice. Proc. Soc. exp. Biol. Med. 1963;112:161–165. doi: 10.3181/00379727-112-27980. [DOI] [PubMed] [Google Scholar]
  • 29.Schochetman G., Stevens R. H., Simpson R. W. Presence of infectious polyadenylated RNA in the coronavirus avian bronchitis virus. Virology. 1977;77:772–782. doi: 10.1016/0042-6822(77)90498-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Sharpee R. L., Mebus C. A., Bass E. P. Characterization of a calf diarrheal coronavirus. Amer. J. vet. Res. 1976;37:1031–1041. [PubMed] [Google Scholar]
  • 31.Sturman L. S. Characterization of a coronavirus. 1. Structural proteins: Effects of preparative conditions on the migration of protein in polyacrylamide gels. Virology. 1977;77:637–649. doi: 10.1016/0042-6822(77)90488-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Tajima N. Morphology of transmissible gastro-enteritis virus of pigs. A possible member of coronaviruses. Arch. ges. Virusforsch. 1970;29:105–108. doi: 10.1007/BF01253886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Tyrrel D. A. J., Almeida J. D., Cunningham C. H., Dowdle W. R., Hofstad M. S., McIntosh K., Tajima M., Zakstelskaya L. Y., Easterday B. C., Kapikian A., Bingham R. W. Coronaviridae. Intervirology. 1975;5:76–82. doi: 10.1159/000149883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal. Biochem. 1969;30:148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES