Skip to main content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Arch Virol. 1990; 114(3): 175–189.
doi: 10.1007/BF01310747
PMCID: PMC7086846
PMID: 2173524

Initial events in bovine coronavirus infection: analysis through immunogold probes and lysosomotropic inhibitors

H. R. Payne,1,2 J. Storz,1 and W. G. Henk3
Author information Article notes Copyright and License information PMC Disclaimer

Summary

The early events in the infection of human rectal tumor cells by bovine coronavirus were investigated by colloidal gold-mediated immunoelectron microscopy and by analysis of the effect of lysosomotropic weak bases on virus yield. Electron microscopic studies revealed sites of fusion between the virus envelope and the plasmalemma but fusion events along intracellular membranes were not observed despite extensive searches. Virion-antibody-colloidal gold complexes were, in fact, endocytosed by synchronously infected cells. These complexes were apparently non-infectious, and they accumulated in vacuoles that resembled secondary lysosomes. Exposure of cells to ammonium chloride or to methylamine during the first hour of infection had little inhibitory effect on the production of infectious virus. Chloroquine treatments were inhibitory but this effect depended on relatively late events in the infectious process. The chloroquine inhibitory step blocked infection of virus adsorbed to cells that were exposed to buffers in the pH range of 4.4 to 8.4. These findings indicate that BCV penetrates its host cell by direct fusion with the plasmalemma and does not require an acidic intracellular compartment for infectious entry.

Keywords: Chloroquine, Ammonium Chloride, Methylamine, Rectal Tumor, Immunoelectron Microscopy

References

1. Andersen KB, Nexo BA. Entry of murine retrovirus into mouse fibroblasts. Virology. 1983;125:85–98. [PubMed] [Google Scholar]
2. Cassell S, Edwards J, Brown DT. Effects of lysosomotropic weak bases on infection of BHK-12 cells by Sindbis virus. J Virol. 1984;52:857–864. [PMC free article] [PubMed] [Google Scholar]
3. Chasey D, Alexander DJ. Morphogenesis of avian infectious bronchitis virus in primary chick kidney cells. Arch Virol. 1976;52:101–111. [PMC free article] [PubMed] [Google Scholar]
4. David-Ferreira JF, Manaker RA. An electron microscope study of the development of a mouse hepatitis virus in tissue culture cells. J Cell Biol. 1965;24:57–78. [PMC free article] [PubMed] [Google Scholar]
5. Dimmock NJ. Initial stages in infection with animal virus. J Gen Virol. 1982;59:1–22. [PubMed] [Google Scholar]
6. Doughri AM, Storz J, Hajer I, Fernando HS. Morphology and morphogenesis of a coronavirus infecting intestinal epithelial cells of newborn calves. Exp Mol Pathol. 1976;25:355–370. [PMC free article] [PubMed] [Google Scholar]
7. Gonzales-Scarano F, Pobjecky N, Nathanson N. La Crosse bunyavirus can mediate pH-dependent fusion from without. Virology. 1984;132:222–225. [PubMed] [Google Scholar]
8. Helenius A, Kartenbeck J, Simons K, Fries E. On the entry of Semliki Forest virus into BHK-21 cells. J Cell Biol. 1980;84:404–420. [PMC free article] [PubMed] [Google Scholar]
9. Helenius A, Marsh M, White J. Inhibition of Semliki Forest virus penetration by lysosomotropic weak bases. J Gen Virol. 1982;58:47–61. [PubMed] [Google Scholar]
10. Holmes KV. Replication of coronaviruses. In: Fields BN, Knipe DM, Chanock RN, Melnick J, Roizman B, Shope R, editors. Virology. New York: Raven Press; 1985. pp. 1331–1343. [Google Scholar]
11. House JA. Economic impact of rotavirus and other neonatal disease agents of animals. J Am Vet Med Assoc. 1978;173:573–576. [PubMed] [Google Scholar]
12. Kielian MC, Keranen S, Kaariainen L, Helenius A. Membrane fusion mutants of Semliki Forest virus. J Cell Biol. 1984;98:139–145. [PMC free article] [PubMed] [Google Scholar]
13. Krzystniak K, Dupuy JM. Early interaction between mouse hepatitis virus 3 and cells. J Gen Virol. 1981;57:53–61. [PubMed] [Google Scholar]
14. Krzystniak K, Dupuy JM. Entry of mouse hepatitis virus 3 into cells. J Gen Virol. 1984;65:227–231. [PubMed] [Google Scholar]
15. Lenard J, Miller KD. Uncoating of enveloped viruses. Cell. 1982;28:5–6. [PubMed] [Google Scholar]
16. Mallucci L. Effect of chloroquine on lysosomes and on growth of mouse hepatitis virus (MHV-3) Virology. 1966;28:355–362. [PubMed] [Google Scholar]
17. Marsh M, Helenius A. Adsorptive endocytosis of Semliki Forest virus. J Mol Biol. 1980;142:439–454. [PubMed] [Google Scholar]
18. Marsh M, Bolzau E, Helenius A. Penetration of Semliki Forest virus from acidic prelysosomal vacuoles. Cell. 1983;32:931–940. [PubMed] [Google Scholar]
19. Marsh M, Matlin K, Simons K, Reggio H, White J, Kartenbeck J, Helenius A. Are lysosomes a site of enveloped-virus penetration? Cold Spring Harbor Symp Quant Biol. 1982;46:835–843. [PubMed] [Google Scholar]
20. Matlin KS, Reggio J, Helenius A, Simons K. Entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981;91:601–613. [PMC free article] [PubMed] [Google Scholar]
21. Maxfield FR. Weak bases and ionophores rapidly and reversibly raise the pH of endocytic vesicles in cultured mouse fibroblasts. J Cell Biol. 1982;95:676–681. [PMC free article] [PubMed] [Google Scholar]
22. Mizzen L, Hilton A, Cheley S, Anderson R. Attenuation of murine coronavirus infection by ammonium chloride. Virology. 1985;142:378–388. [PMC free article] [PubMed] [Google Scholar]
23. Mollenhauer HH. Plastic embedding mixtures for use in electron microscopy. Stain Technol. 1964;39:111–114. [PubMed] [Google Scholar]
24. Nemerow GR, Cooper NR. Early events in the infection of human B lymphocytes by Epstein-Barr virus: the internalization process. Virology. 1984;132:186–198. [PubMed] [Google Scholar]
25. Okhuma S, Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci USA. 1978;75:3327–3331. [PMC free article] [PubMed] [Google Scholar]
26. Patterson S, Bingham RW. Electron microscope observations on the entry of avain infectious bronchitis virus into susceptible cells. Arch Virol. 1976;53:267–273. [PMC free article] [PubMed] [Google Scholar]
27. Payne HR, Storz J. Analysis of cell fusion induced by bovine coronavirus infection. Arch Virol. 1988;103:27–33. [PMC free article] [PubMed] [Google Scholar]
28. Poste G, Pasternak CA. Virus-induced cell fusion. In: Poste G, Nicholson GL, editors. Cell surface reviews, vol 4. Amsterdam: Elsevier/North-Holland; 1978. pp. 305–357. [Google Scholar]
29. Sharpee RL, Mebus CA, Bass EP. Characterization of a calf diarrheal coronavirus. Am J Vet Res. 1976;37:1031–1041. [PubMed] [Google Scholar]
30. Siddell SG, Wege H, ter Meulen V. The structure and replication of coronaviruses. Curr Top Microbiol Immunol. 1982;99:131–163. [PubMed] [Google Scholar]
31. Siddell SG, Anderson R, Cavanagh D, Fujiwara K, Klenk HD, Macnaughton MR, Pensaert M, Stohlman SA, Sturman L, van der Zeijst BAM. Coronaviridae. Intervirology. 1983;20:181–189. [PMC free article] [PubMed] [Google Scholar]
32. Sturman LS, Holmes KV. The molecular biology of coronaviruses. Adv Virus Res. 1982;28:35–112. [PMC free article] [PubMed] [Google Scholar]
33. Sturman LS, Ricard CS, Holmes KV. Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: activation of cell-fusion activity of virions by trypsin and separation of two different 90 k cleavage fragments. J Virol. 1985;56:904–911. [PMC free article] [PubMed] [Google Scholar]
33a. Sturman LS, Ricard CS, Holmes KV. Conformational change of the coronavirus peplomer glycoprotein at pH 8.0 and 37°C correlates with virus aggregation and virus-induced cell fusion. J Virol. 1990;64:3042–3050. [PMC free article] [PubMed] [Google Scholar]
34. St Cyr-Coats K, Storz J, Hussain KA, Schnorr KL. Structural proteins of bovine coronavirus strain L9: effects of the host cells and trypsin treatment. Arch Virol. 1988;103:35–43. [PMC free article] [PubMed] [Google Scholar]
35. Tompkins WAF, Watrach AM, Schmale JD, Schultz RM, Harris JA. Cultural and antigenic properties of newly established cell strains from adenocarcinomas of human colon and rectum. J Natl Cancer Inst. 1974;52:101–106. [PubMed] [Google Scholar]
36. Tooze J, Tooze SA. Infection of AtT20 murine pituitary tumour cells by mouse hepatitis virus strain A59: virus budding is restricted to the Golgi region. EMBO J. 1985;37:203–212. [PubMed] [Google Scholar]
37. Tycko B, Maxfield FR. Rapid acidification of endocytic vesicles containing alpha-2-macroglobulin. Cell. 1982;28:643–651. [PubMed] [Google Scholar]
38. Vlasak R, Lnytzes 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. [PMC free article] [PubMed] [Google Scholar]
38a. Weismiller DG, Sturman LS, Buchmeier MJ, Fleming JO, Holmes KV. Monoclonal antibodies to the peplomer glycoprotein of coronavirus mouse hepatitis virus identify two subunits and detect a conformational change in the subunit released under mild alkaline conditions. J Virol. 1990;64:3051–3055. [PMC free article] [PubMed] [Google Scholar]
39. White J, Kartenbeck J, Helenius A. Fusion of Semliki forest virus with the plasma membrane can be induced by low pH. J Cell Biol. 1980;87:264–272. [PMC free article] [PubMed] [Google Scholar]
40. White J, Matlin K, Helenius A. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol. 1981;89:674–679. [PMC free article] [PubMed] [Google Scholar]
Articles from Archives of Virology are provided here courtesy of Nature Publishing Group
-