Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1988 Dec;62(12):4686–4690. doi: 10.1128/jvi.62.12.4686-4690.1988

The E3 protein of bovine coronavirus is a receptor-destroying enzyme with acetylesterase activity.

R Vlasak 1, W Luytjes 1, J Leider 1, W Spaan 1, P Palese 1
PMCID: PMC253582  PMID: 3184275

Abstract

In addition to members of the Orthomyxoviridae and Paramyxoviridae, several coronaviruses have been shown to possess receptor-destroying activities. Purified bovine coronavirus (BCV) preparations have an esterase activity which inactivates O-acetylsialic acid-containing receptors on erythrocytes. Diisopropyl fluorophosphate (DFP) completely inhibits this receptor-destroying activity of BCV, suggesting that the viral enzyme is a serine esterase. Treatment of purified BCV with [3H]DFP and subsequent sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the proteins revealed that the E3 protein was specifically phosphorylated. This finding suggests that the esterase/receptor-destroying activity of BCV is associated with the E3 protein. Furthermore, treatment of BCV with DFP dramatically reduced its infectivity in a plaque assay. It is assumed that the esterase activity of BCV is required in an early step of virus replication, possibly during virus entry or uncoating.

Full text

PDF
4686

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BENDER M. L., KEZDY J. MECHANISM OF ACTION OF PROTEOLYTIC ENZYMES. Annu Rev Biochem. 1965;34:49–76. doi: 10.1146/annurev.bi.34.070165.000405. [DOI] [PubMed] [Google Scholar]
  2. Brand C., Palese P. Sequential passage of influenza virus in embryonated eggs or tissue culture: emergence of mutants. Virology. 1980 Dec;107(2):424–433. doi: 10.1016/0042-6822(80)90309-8. [DOI] [PubMed] [Google Scholar]
  3. Deregt D., Sabara M., Babiuk L. A. Structural proteins of bovine coronavirus and their intracellular processing. J Gen Virol. 1987 Nov;68(Pt 11):2863–2877. doi: 10.1099/0022-1317-68-11-2863. [DOI] [PubMed] [Google Scholar]
  4. Formanowski F., Meier-Ewert H. Isolation of the influenza C virus glycoprotein in a soluble form by bromelain digestion. Virus Res. 1988 May;10(2-3):177–191. doi: 10.1016/0168-1702(88)90014-7. [DOI] [PubMed] [Google Scholar]
  5. GOTTSCHALK A. Neuraminidase: the specific enzyme of influenza virus and Vibrio cholerae. Biochim Biophys Acta. 1957 Mar;23(3):645–646. doi: 10.1016/0006-3002(57)90389-x. [DOI] [PubMed] [Google Scholar]
  6. Herrler G., Dürkop I., Becht H., Klenk H. D. The glycoprotein of influenza C virus is the haemagglutinin, esterase and fusion factor. J Gen Virol. 1988 Apr;69(Pt 4):839–846. doi: 10.1099/0022-1317-69-4-839. [DOI] [PubMed] [Google Scholar]
  7. Herrler G., Rott R., Klenk H. D., Müller H. P., Shukla A. K., Schauer R. The receptor-destroying enzyme of influenza C virus is neuraminate-O-acetylesterase. EMBO J. 1985 Jun;4(6):1503–1506. doi: 10.1002/j.1460-2075.1985.tb03809.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Homma M., Ouchi M. Trypsin action on the growth of Sendai virus in tissue culture cells. 3. Structural difference of Sendai viruses grown in eggs and tissue culture cells. J Virol. 1973 Dec;12(6):1457–1465. doi: 10.1128/jvi.12.6.1457-1465.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. King B., Brian D. A. Bovine coronavirus structural proteins. J Virol. 1982 May;42(2):700–707. doi: 10.1128/jvi.42.2.700-707.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. King B., Potts B. J., Brian D. A. Bovine coronavirus hemagglutinin protein. Virus Res. 1985 Feb;2(1):53–59. doi: 10.1016/0168-1702(85)90059-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Luytjes W., Sturman L. S., Bredenbeek P. J., Charite J., van der Zeijst B. A., Horzinek M. C., Spaan W. J. Primary structure of the glycoprotein E2 of coronavirus MHV-A59 and identification of the trypsin cleavage site. Virology. 1987 Dec;161(2):479–487. doi: 10.1016/0042-6822(87)90142-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Muchmore E. A., Varki A. Selective inactivation of influenza C esterase: a probe for detecting 9-O-acetylated sialic acids. Science. 1987 Jun 5;236(4806):1293–1295. doi: 10.1126/science.3589663. [DOI] [PubMed] [Google Scholar]
  13. Nagai Y., Klenk H. D. Activation of precursors to both glycoporteins of Newcastle disease virus by proteolytic cleavage. Virology. 1977 Mar;77(1):125–134. doi: 10.1016/0042-6822(77)90412-3. [DOI] [PubMed] [Google Scholar]
  14. Ohuchi M., Ohuchi R., Mifune K. Demonstration of hemolytic and fusion activities of influenza C virus. J Virol. 1982 Jun;42(3):1076–1079. doi: 10.1128/jvi.42.3.1076-1079.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Palese P., Schulman J. Isolation and characterization of influenza virus recombinants with high and low neuraminidase activity. Use of 2-(3'-methoxyphenyl)-n-acetylneuraminic acid to identify cloned populations. Virology. 1974 Jan;57(1):227–237. doi: 10.1016/0042-6822(74)90123-8. [DOI] [PubMed] [Google Scholar]
  16. Palese P., Tobita K., Ueda M., Compans R. W. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase. Virology. 1974 Oct;61(2):397–410. doi: 10.1016/0042-6822(74)90276-1. [DOI] [PubMed] [Google Scholar]
  17. Parvin J. D., Moscona A., Pan W. T., Leider J. M., Palese P. Measurement of the mutation rates of animal viruses: influenza A virus and poliovirus type 1. J Virol. 1986 Aug;59(2):377–383. doi: 10.1128/jvi.59.2.377-383.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rogers G. N., Herrler G., Paulson J. C., Klenk H. D. Influenza C virus uses 9-O-acetyl-N-acetylneuraminic acid as a high affinity receptor determinant for attachment to cells. J Biol Chem. 1986 May 5;261(13):5947–5951. [PubMed] [Google Scholar]
  19. Scheid A., Caliguiri L. A., Compans R. W., Choppin P. W. Isolation of paramyxovirus glycoproteins. Association of both hemagglutinating and neuraminidase activities with the larger SV5 glycoprotein. Virology. 1972 Dec;50(3):640–652. doi: 10.1016/0042-6822(72)90418-7. [DOI] [PubMed] [Google Scholar]
  20. Scheid A., Choppin P. W. Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis, and infectivity of proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology. 1974 Feb;57(2):475–490. doi: 10.1016/0042-6822(74)90187-1. [DOI] [PubMed] [Google Scholar]
  21. Scheid A., Choppin P. W. The hemagglutinating and neuraminidase protein of a paramyxovirus: interaction with neuraminic acid in affinity chromatography. Virology. 1974 Nov;62(1):125–133. doi: 10.1016/0042-6822(74)90308-0. [DOI] [PubMed] [Google Scholar]
  22. Spaan W. J., Rottier P. J., Horzinek M. C., van der Zeijst B. A. Isolation and identification of virus-specific mRNAs in cells infected with mouse hepatitis virus (MHV-A59). Virology. 1981 Jan 30;108(2):424–434. doi: 10.1016/0042-6822(81)90449-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sturman L. S., Holmes K. V. 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]
  24. Sturman L. S., Ricard C. S., Holmes K. V. 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 Dec;56(3):904–911. doi: 10.1128/jvi.56.3.904-911.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Talbot P. J., Salmi A. A., Knobler R. L., Buchmeier M. J. Topographical mapping of epitopes on the glycoproteins of murine hepatitis virus-4 (strain JHM): correlation with biological activities. Virology. 1984 Jan 30;132(2):250–260. doi: 10.1016/0042-6822(84)90032-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vlasak R., Krystal M., Nacht M., Palese P. The influenza C virus glycoprotein (HE) exhibits receptor-binding (hemagglutinin) and receptor-destroying (esterase) activities. Virology. 1987 Oct;160(2):419–425. doi: 10.1016/0042-6822(87)90013-4. [DOI] [PubMed] [Google Scholar]
  27. Vlasak R., Luytjes W., Spaan W., Palese P. Human and bovine coronaviruses recognize sialic acid-containing receptors similar to those of influenza C viruses. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4526–4529. doi: 10.1073/pnas.85.12.4526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wege H., Wege H., Nagashima K., ter Meulen V. Structural polypeptides of the murine coronavirus JHM. J Gen Virol. 1979 Jan;42(1):37–47. doi: 10.1099/0022-1317-42-1-37. [DOI] [PubMed] [Google Scholar]
  29. Wiley D. C., Skehel J. J. The structure and function of the hemagglutinin membrane glycoprotein of influenza virus. Annu Rev Biochem. 1987;56:365–394. doi: 10.1146/annurev.bi.56.070187.002053. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES