1887

Abstract

Summary

Using [H]glucosamine and [H]mannose labels, two virus-specific glycosylated polypeptide species with values of about 200000 (200K) and in the 75K to 100K range, respectively, were recognized in Berne virus-infected embryonic mule skin cells. In purified virions only the latter glycoprotein occurred. Concanavalin A was bound to the virion as evidenced by reduction in infectivity. Analyses using SDS-PAGE, blotting and glycoprotein identification with concanavalin A and horseradish peroxidase showed coincidence of the virion glycoprotein signals with the maximum infectivity and haemagglutinating activity in an isokinetic sucrose gradient. Polyclonal rabbit immune serum and a neutralizing and haemagglutination-inhibiting monoclonal antibody raised against Berne virus recognized both the 75K to 100K and the ‘200K’ glycoproteins. Using tunicamycin, a concentration-dependent inhibition of infectivity was noted; however, non-infectious particles containing the two major polypeptides (20K and 22K) were released from the cells in small quantities. The glycoproteins were absent from cytoplasmic extracts and a novel polypeptide of about 150K was identified instead. Translation of poly(A)-selected intracellular RNA from infected cells in a rabbit reticulocyte cell-free system also resulted in the appearance of a new high polypeptide (about 170K). Using pulse-chase labelling and radioimmunoprecipitation, suggestive evidence for a precursor-product relationship between the intracellular ‘200K’ and the virion glycoproteins has been obtained. These experiments identify the -glycosylated proteins in the 75K to 100K range as constituents of the peplomeric envelope projection of Berne virus; they probably arise by post-translational processing of a 150K to 170K precursor molecule involving glycosylation and subsequent cleavage.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-67-11-2475
1986-11-01
2021-10-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/67/11/JV0670112475.html?itemId=/content/journal/jgv/10.1099/0022-1317-67-11-2475&mimeType=html&fmt=ahah

References

  1. Clegg J. C. S. 1982; Glycoprotein detection in nitrocellulose transfers of electrophoretically separated protein mixtures using concanavalin A and peroxidase : application to arenavirus and flavivirus proteins. Analytical Biochemistry 127:389–394
    [Google Scholar]
  2. Garwes D. J., Lucas M. H., Higgins D. A., Pike B. V., Cartwright S. F. 1979; Antigenicity of structural components from porcine transmissible gastroenteritis virus. Veterinary Microbiology 3:179–190
    [Google Scholar]
  3. Glass W. F., Briggs R. C., Hnlica L. S. 1981; Use of lectins for detection of electrophoretically separated glycoproteins transferred onto nitrocellulose sheets. Analytical Biochemistry 115:219–224
    [Google Scholar]
  4. Horzinek M. C., Weiss M. 1984; Toroviridae: a taxonomic proposal. Zentralblatt für Veterinärmedizin B 31:649–659
    [Google Scholar]
  5. Horzinek M. C., Lutz H., Pedersen N. C. 1982; Antigenic relationships among homologous structural polypeptides of porcine, feline and canine coronaviruses. Infection and Immunity 37:1148–1155
    [Google Scholar]
  6. Horzinek M. C., Weiss M., Ederveen J. 1984; Berne virus is not ‘coronavirus-like’. Journal of General Virology 65:645–649
    [Google Scholar]
  7. Horzinek M. C., Ederveen J., Weiss M. 1985; The nucleocapsid of Berne virus. Journal of General Virology 66:1287–1296
    [Google Scholar]
  8. Jackson R. J., Hunt T. 1983; Preparation and use of nuclease treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA. Methods in Enzymology 96:50–74
    [Google Scholar]
  9. Koolen M. J. M., Osterhaus A. D. M. E., Siebelink K. H. J., Horzinek M. C., Van Der Zeijst B. A. M. 1984; Monoclonal antibodies to the three classes of mouse hepatitis strain A59 proteins. Advances in Experimental Medicine and Biology 173:115–116
    [Google Scholar]
  10. Koopmans M., Ederveen J., Woode G. N., Horzinek M. C. 1986; The surface proteins of Breda virus. American Journal of Veterinary Research (in press)
    [Google Scholar]
  11. Mockett A. P. A., Cavanagh D., Brown T. D. K. 1984; Monoclonal antibodies to the SI spike and membrane proteins of avian infectious bronchitis coronavirus strain Massachusetts M41. Journal of General Virology 65:2281–2286
    [Google Scholar]
  12. Rottier P. J. M., Spaan W. J. M., Horzinek M. C., Van Der Zeijst B. A. M. 1981; Translation of three mousehepatitis virus strain A59 subgenomic RNAs in Xenopus laevis oocytes. Journal of Virology 38:20–26
    [Google Scholar]
  13. Talbot P. J., Salmi A. A., Knobler R. L., Buchmeier M. J. 1984; Topographical mapping of epitopes on the glycoprotein of murine hepatitis virus-4 (strain JHM): correlation with biological activities. Virology 132:250–260
    [Google Scholar]
  14. Van der zeijst B. A. M., Bloemers H. P. J. 1976; 4698 Isokinetic glycerol and sucrose gradients for density gradient centrifugation. In Handbook of Biochemistry and Molecular Biology 3rd edn vol 1 pp 426–519 Edited by Fasman G. D. Cleveland: CRC Press;
    [Google Scholar]
  15. Wege H., Dörries R., Wege H. 1984; Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes of the peplomer protein (E2). Journal of General Virology 65:1931–1942
    [Google Scholar]
  16. Weiss M., Horzinek M. C. 1986a; Morphogenesis of Berne virus (proposed family Toroviridae). Journal of General Virology 67:1305–1314
    [Google Scholar]
  17. Weiss M., Horzinek M. C. 1986b; Resistance of Berne virus to physical and chemical treatment. Veterinary Microbiology 11:41–49
    [Google Scholar]
  18. Weiss M., Horzinek M. C. 1986c; The proposed family Toroviridae: agents of enteric infections. Archives of Virology (in press)
    [Google Scholar]
  19. Weiss M., Steck F., Horzinek M. C. 1983; Purification and partial characterization of a new enveloped RNA virus (Berne virus). Journal of General Virology 64:1849–1858
    [Google Scholar]
  20. Woode G. N., Reed D. E., Runnels P. L., Herrig M. A., Hill H. T. 1982; Studies with an unclassified virus isolated from diarrheic calves. Veterinary Microbiology 7:221–240
    [Google Scholar]
  21. Zanoni R., Weiss M., Peterhans E. 1986; The haemagglutinating activity of Berne virus. Journal of General Virology 67:2485–2488
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-67-11-2475
Loading
/content/journal/jgv/10.1099/0022-1317-67-11-2475
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error