Skip to content
1887

Journal of General Virology header logo Journal of General Virology

Volume 85, Issue 5

The surface glycoprotein E2 of bovine viral diarrhoea virus contains an intracellular localization signal No Access

Abstract

The intracellular transport of the surface glycoprotein E2 of bovine viral diarrhoea virus was analysed by expressing the cloned gene in the absence of other viral proteins. Immunofluorescence analysis and surface biotinylation indicated that E2 is located in an early compartment of the secretory pathway and not transported to the cell surface. In agreement with this result, E2 was found to contain only high-mannose oligosaccharide side-chains but no -glycans of the complex type. To define the intracellular localization signal of the E2 protein, chimeric proteins were generated. E2 chimeras containing the MT (membrane anchor plus carboxy-terminal domain) of the G protein of vesicular stomatitis virus (VSV) or of the F protein of bovine respiratory syncytial virus (BRSV) were transported to the cell surface. On the other hand, VSV G protein containing the MT domain of E2 was detected only in the ER, indicating that this domain contains an ER localization signal. A chimeric E2 protein, in which not the membrane anchor but only the carboxy-terminal end was replaced by the corresponding domain of the BRSV F protein, was also localized in the ER. Therefore, it was concluded that the membrane anchor contains the ER localization signal of E2. Interestingly, the ER export signal within the VSV G protein cytoplasmic tail was found to overrule the ER localization signal in the E2 protein membrane anchor.

  • Received:
  • Accepted:
  • Published Online:
SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.19740-0
2004-05-01
2025-03-23
Loading full text...

Full text loading...

References

  1. Allison S. L., Stiasny K., Stadler K., Mandl C. W., Heinz F. X. 1999; Mapping of functional elements in the stem-anchor region of tick-borne encephalitis virus envelope protein E. J Virol 73:5605–5612
     [Google Scholar]
  2. Baker J. C. 1995; The clinical manifestations of bovine viral diarrhea infection. Vet Clin North Am Food Anim Pract 11:425–445
     [Google Scholar]
  3. Bartosch B., Dubuisson J., Cosset F. L. 2003; Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J Exp Med 197:633–642
     [Google Scholar]
  4. Bolin S. R., McClurkin A. W., Coria M. F. 1985; Frequency of persistent bovine viral diarrhea virus infection in selected cattle herds. Am J Vet Res 46:2385–2387
     [Google Scholar]
  5. Bolin S. R., Moennig V., Kelso-Gourley V., Ridpath J. 1988; Monoclonal antibodies with neutralising activity segregate isolates of bovine viral diarrhea virus into groups. Arch Virol 99:117–123
     [Google Scholar]
  6. Bonifacino J. S., Cosson P., Shah N., Klausner R. D. 1991; Role of potentially charged transmembrane residues in targeting proteins for retention and degradation within the endoplasmic reticulum. EMBO J 10:2783–2793
     [Google Scholar]
  7. Buchholz U. J., Finke S., Conzelmann K.-K. 1999; Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for virus replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter. J Virol 73:251–259
     [Google Scholar]
  8. Cocquerel L., Meunier J. C., Pillez A., Wychowski C., Dubuisson J. 1998; A retention signal necessary and sufficient for endoplasmic reticulum localisation maps to the transmembrane domain of hepatitis C virus glycoprotein E2. J Virol 72:2183–2191
     [Google Scholar]
  9. Cocquerel L., Wychowski C., Minner F., Penin F., Dubuisson J. 2000; Charged residues in the transmembrane domains of hepatitis C virus glycoproteins play a major role in the processing, subcellular localisation, and assembly of these envelope proteins. J Virol 74:3623–3633
     [Google Scholar]
  10. Cocquerel L., Op de Beeck A., Lambot M., Roussel J., Delgrange D., Pillez A., Wychowski C., Penin F., Dubuisson J. 2002; Topological changes in the transmembrane domains of hepatitis C virus envelope glycoproteins. EMBO J 21:2893–2902
     [Google Scholar]
  11. Compans R. W., Herrler G. 1999; Epithelial cells and viral infection. In Mucosal Immunology pp  671–683 Edited by Ogra P. L., Mestecky J., Lamm M. E., Strober W., Bienenstock J., McGhee J. R. New York: Academic Press;
     [Google Scholar]
  12. Dalbey R. E. 1990; Positively charged residues are important determinants of membrane topology. Trends Biochem Sci 15:253–257
     [Google Scholar]
  13. Donis R. O., Corapi W. V., Dubovi E. J. 1988; Neutralizing monoclonal antibodies to bovine viral diarrhoea virus bind to the 56K to 58K glycoprotein. J Gen Virol 69:77–86
     [Google Scholar]
  14. Drummer H. E., Maerz A., Poumbourios P. 2003; Cell surface expression of functional hepatitis C virus E1 and E2 glycoproteins. FEBS Lett 546:385–390
     [Google Scholar]
  15. Elbers K., Tautz N., Becher P., Stoll D., Rumenapf T., Thiel H. J. 1996; Processing in the pestivirus E2-NS2 region: identification of proteins p7 and E2p7. J Virol 70:4131–4135
     [Google Scholar]
  16. Gray E. W., Nettleton P. F. 1987; The ultrastructure of cell cultures infected with border disease and bovine virus diarrhoea viruses. J Gen Virol 68:2339–2346
     [Google Scholar]
  17. Greiser-Wilke I., Dittmar K. E., Liess B., Moennig V. 1991; Immunofluorescence studies of biotype-specific expression of bovine viral diarrhoea virus epitopes in infected cells. J Gen Virol 72:2015–2019
     [Google Scholar]
  18. Grigera P. R., Marzocca M. P., Capozzo A. V. E., Buonocore L., Donis R. O., Rose J. K. 2000; Presence of bovine viral diarrhea virus (BVDV) E2 glycoprotein in VSV recombinant particles and induction of neutralizing BVDV antibodies in mice. Virus Res 69:3–15
     [Google Scholar]
  19. Grummer B., Beer M., Liebler-Tenorio E., Greiser-Wilke I. 2001; Localisation of viral proteins in cells infected with bovine viral diarrhoea virus. J Gen Virol 82:2597–2605
     [Google Scholar]
  20. Harada T., Tautz N., Thiel H. J. 2000; E2-p7 region of the bovine viral diarrhea virus polyprotein: processing and functional studies. J Virol 74:9498–9506
     [Google Scholar]
  21. Hsu M., Zhang J., Flint M., Logvinoff C., Cheng-Mayer C., Rice C. M., McKeating J. A. 2003; Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad U S A 100:7271–7276
     [Google Scholar]
  22. Kweon C.-H., Yoon Y.-D., An S.-H., Lee Y.-S. 1997; Expression of envelope protein (E2) of bovine viral diarrhea virus in insect cells. J Vet Med Sci 59:415–419
     [Google Scholar]
  23. Magar R., Minocha H. C., Lecomte J. 1988; Bovine viral diarrhea virus proteins: heterogeneity of cytopathogenic and noncytopathogenic strains and evidence of a 53K glycoprotein neutralization epitope. Vet Microbiol 16:303–314
     [Google Scholar]
  24. Meyers G., Thiel H. J. 1996; Molecular characterisation of pestiviruses. Adv Virus Res 47:53–118
     [Google Scholar]
  25. Monne M., Nilsson I., Elofsson A., von Heijne G. 1999; Turns in transmembrane helices: determination of the minimal length of a “helical hairpin” and derivation of a fine-grained turn propensity scale. J Mol Biol 293:807–814
     [Google Scholar]
  26. Moss B., Elroy-Stein O., Mizukami T., Alexander W. A., Fuerst T. R. 1990; Product review. New mammalian expression vectors. Nature 348:91–92
     [Google Scholar]
  27. Nishimura N., Balch W. E. 1997; A di-acidic signal required for selective export from the endoplasmic reticulum. Science 277:556–558
     [Google Scholar]
  28. Op de Beeck A., Dubuisson J. 2003; Topology of hepatitis C virus envelope glycoproteins. Rev Med Virol 13:233–241
     [Google Scholar]
  29. Orban S., Liess B., Hafez S. M., Frey H.-R., Blindow H., Sasse-Patzer B. 1983; Studies on transplacental transmissibility of bovine virus diarrhoea (BVD) vaccine virus. I. Inoculation of pregnant cows 15 to 90 days before parturition (190th to 265th day of gestation). Zentbl Vetmed Reihe B 30:619–634
     [Google Scholar]
  30. Rümenapf T., Unger G., Strauss J. H., Thiel H.-J. 1993; Processing of the envelope glycoproteins of pestiviruses. J Virol 67:3288–3294
     [Google Scholar]
  31. Sevier C. S., Weisz O. A., Davis M., Machamer C. E. 2000; Efficient export of the vesicular stomatitis virus G protein from the endoplasmic reticulum requires a signal in the cytoplasmic tail that includes both tyrosine-based and di-acidic motifs. Mol Biol Cell 11:13–22
     [Google Scholar]
  32. Weiland F., Weiland E., Unger G., Saalmuller A., Thiel H. J. 1999; Localisation of pestiviral envelope proteins ERNS and E2 at the cell surface and on isolated particles. J Gen Virol 80:1157–1165
     [Google Scholar]
  33. Wengler G., Bradley D. W., Collett M. S., Heinz F. X., Schlesinger R. W., Strauss J. H. 1995; Flaviviridae . In Virus Taxonomy. Sixth Report of the International Commitee on Taxonomy of Viruses pp  415–427 Edited by Murphy F. A., Fauquet C. M., Bishop D. H. L., Ghabrial S. A., Jarvis A. W., Martelli G. P., Mayo M. A., Summers M. D. New York: Springer;
     [Google Scholar]
  34. Zhang W., Chipman P. R., Corver J. 7 other authors 2003; Visualization of membrane protein domains by cryo-electron microscopy of dengue virus. Nat Struct Biol 10:907–912
     [Google Scholar]
  35. Zhou Y., König M., Hobom G., Neumeier E. 1998; Membrane-anchored incorporation of a foreign protein in recombinant influenza virions. Virology 246:83–94
     [Google Scholar]
  36. Zimmer G., Budz L., Herrler G. 2001a; Proteolytic activation of respiratory syncytial virus fusion protein. Cleavage at two furin consensus sequences. J Biol Chem 276:31642–31650
     [Google Scholar]
  37. Zimmer G., Trotz I., Herrler G. 2001b; N-glycans of F protein differentially affect fusion activity of human respiratory syncytial virus. J Virol 75:4744–4751
     [Google Scholar]
  38. Zimmer G., Conzelmann K.-K., Herrler G. 2002; Cleavage at the furin consensus sequence RAR/KR(109) and presence of the intervening peptide of the respiratory syncytial virus fusion protein are dispensable for virus replication in cell culture. J Virol 76:9218–9224
     [Google Scholar]
/content/journal/jgv/10.1099/vir.0.19740-0
Loading
The surface glycoprotein E2 of bovine viral diarrhoea virus contains an intracellular localization signal
J. Gen. Virol. 85, 1101 (2004); https://doi.org/10.1099/vir.0.19740-0
/content/journal/jgv/10.1099/vir.0.19740-0
/content/journal/jgv/10.1099/vir.0.19740-0
Loading

Data & Media loading...

Most cited 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