1887

Abstract

Glycoprotein B (gB) homologues within the herpesvirus family display high sequence conservation, and a number of gB homologues contain a cleavage motif R-X-K/R-R recognized by the cellular protease furin. Epstein–Barr virus (EBV) gB contains this motif and cleaved gB is found in EBV virions. To determine the functional significance of this cleavage motif in EBV gB, a deletion mutant (gB Δfurin) was created lacking the motif. This cleavage mutant was expressed well in cell culture but was not cleaved. Experiments examining gB Δfurin in a cell-fusion assay revealed that fusion was reduced by 52 % in epithelial and 28 % in B cells when compared with wild-type EBV gB. This decrease in cell–cell fusion is similar to that observed with multiple alphaherpesvirus gB cleavage mutants and supports a conserved function for cleaved gB.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.007237-0
2009-03-01
2019-11-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/3/591.html?itemId=/content/journal/jgv/10.1099/vir.0.007237-0&mimeType=html&fmt=ahah

References

  1. Backovic, M., Leser, G. P., Lamb, R. A., Longnecker, R. & Jardetzky, T. S. ( 2007; ). Characterization of EBV gB indicates properties of both class I and class II viral fusion proteins. Virology 368, 102–113.[CrossRef]
    [Google Scholar]
  2. Baghian, A., Luftig, M., Black, J. B., Meng, Y. X., Pau, C. P., Voss, T., Pellett, P. E. & Kousoulas, K. G. ( 2000; ). Glycoprotein B of human herpesvirus 8 is a component of the virion in a cleaved form composed of amino- and carboxyl-terminal fragments. Virology 269, 18–25.[CrossRef]
    [Google Scholar]
  3. Britt, W. J. & Vugler, L. G. ( 1989; ). Processing of the gp55–116 envelope glycoprotein complex (gB) of human cytomegalovirus. J Virol 63, 403–410.
    [Google Scholar]
  4. Burnette, W. N. ( 1981; ). “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate–polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem 112, 195–203.[CrossRef]
    [Google Scholar]
  5. Daniels, G. M. & Amara, S. G. ( 1998; ). Selective labeling of neurotransmitter transporters at the cell surface. Methods Enzymol 296, 307–318.
    [Google Scholar]
  6. Emini, E. A., Luka, J., Armstrong, M. E., Keller, P. M., Ellis, R. W. & Pearson, G. R. ( 1987; ). Identification of an Epstein–Barr virus glycoprotein which is antigenically homologous to the varicella-zoster virus glycoprotein II and the herpes simplex virus glycoprotein B. Virology 157, 552–555.[CrossRef]
    [Google Scholar]
  7. Fleckenstein, B., Muller, I. & Collins, J. ( 1982; ). Cloning of the complete human cytomegalovirus genome in cosmids. Gene 18, 39–46.[CrossRef]
    [Google Scholar]
  8. Gong, M. & Kieff, E. ( 1990; ). Intracellular trafficking of two major Epstein–Barr virus glycoproteins, gp350/220 and gp110. J Virol 64, 1507–1516.
    [Google Scholar]
  9. Gong, M., Ooka, T., Matsuo, T. & Kieff, E. ( 1987; ). Epstein–Barr virus glycoprotein homologous to herpes simplex virus gB. J Virol 61, 499–508.
    [Google Scholar]
  10. Haan, K. M., Lee, S. K. & Longnecker, R. ( 2001; ). Different functional domains in the cytoplasmic tail of glycoprotein B are involved in Epstein–Barr virus-induced membrane fusion. Virology 290, 106–114.[CrossRef]
    [Google Scholar]
  11. Hampl, H., Ben-Porat, T., Ehrlicher, L., Habermehl, K. O. & Kaplan, A. S. ( 1984; ). Characterization of the envelope proteins of pseudorabies virus. J Virol 52, 583–590.
    [Google Scholar]
  12. Hutt-Fletcher, L. M. ( 2007; ). Epstein–Barr virus entry. J Virol 81, 7825–7832.[CrossRef]
    [Google Scholar]
  13. Imai, S., Nishikawa, J. & Takada, K. ( 1998; ). Cell-to-cell contact as an efficient mode of Epstein–Barr virus infection of diverse human epithelial cells. J Virol 72, 4371–4378.
    [Google Scholar]
  14. Johannsen, E., Luftig, M., Chase, M. R., Weicksel, S., Cahir-McFarland, E., Illanes, D., Sarracino, D. & Kieff, E. ( 2004; ). Proteins of purified Epstein–Barr virus. Proc Natl Acad Sci U S A 101, 16286–16291.[CrossRef]
    [Google Scholar]
  15. Kieff, E. & Rickinson, A. B. ( 2007; ). Epstein–Barr virus and its replication. In Fields Virology, 5th edn, vol. 2, pp. 2603–2654. Edited by D. M. Knipe, P. M. Howley, D. E. Griffin, R. A. Lamb, M. A. Martin, B. Roizman & S. E. Straus. Philadelphia, PA: Lippincott Williams & Wilkins.
  16. Kirschner, A. N., Omerovic, J., Popov, B., Longnecker, R. & Jardetzky, T. S. ( 2006; ). Soluble Epstein–Barr virus glycoproteins gH, gL, and gp42 form a 1 : 1 : 1 stable complex that acts like soluble gp42 in B-cell fusion but not in epithelial cell fusion. J Virol 80, 9444–9454.[CrossRef]
    [Google Scholar]
  17. Kopp, A., Blewett, E., Misra, V. & Mettenleiter, T. C. ( 1994; ). Proteolytic cleavage of bovine herpesvirus 1 (BHV-1) glycoprotein gB is not necessary for its function in BHV-1 or pseudorabies virus. J Virol 68, 1667–1674.
    [Google Scholar]
  18. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  19. Lee, S. K., Compton, T. & Longnecker, R. ( 1997; ). Failure to complement infectivity of EBV and HSV-1 glycoprotein B (gB) deletion mutants with gBs from different human herpesvirus subfamilies. Virology 237, 170–181.[CrossRef]
    [Google Scholar]
  20. Loh, L. C. ( 1991; ). Synthesis and processing of the major envelope glycoprotein of murine cytomegalovirus. Virology 180, 239–250.[CrossRef]
    [Google Scholar]
  21. McShane, M. P. & Longnecker, R. ( 2004; ). Cell-surface expression of a mutated Epstein–Barr virus glycoprotein B allows fusion independent of other viral proteins. Proc Natl Acad Sci U S A 101, 17474–17479.[CrossRef]
    [Google Scholar]
  22. McShane, M. P. & Longnecker, R. ( 2005; ). Analysis of fusion using a virus-free cell fusion assay. Methods Mol Biol 292, 187–196.
    [Google Scholar]
  23. Meredith, D. M., Stocks, J. M., Whittaker, G. R., Halliburton, I. W., Snowden, B. W. & Killington, R. A. ( 1989; ). Identification of the gB homologues of equine herpesvirus types 1 and 4 as disulphide-linked heterodimers and their characterization using monoclonal antibodies. J Gen Virol 70, 1161–1172.[CrossRef]
    [Google Scholar]
  24. Okazaki, K. ( 2007; ). Proteolytic cleavage of glycoprotein B is dispensable for in vitro replication, but required for syncytium formation of pseudorabies virus. J Gen Virol 88, 1859–1865.[CrossRef]
    [Google Scholar]
  25. Omerovic, J., Lev, L. & Longnecker, R. ( 2005; ). The amino terminus of Epstein–Barr virus glycoprotein gH is important for fusion with epithelial and B cells. J Virol 79, 12408–12415.[CrossRef]
    [Google Scholar]
  26. Pereira, L. ( 1994; ). Function of glycoprotein B homologues of the family Herpesviridae. Infect Agents Dis 3, 9–28.
    [Google Scholar]
  27. Qualtiere, L. F. & Pearson, G. R. ( 1979; ). Epstein–Barr virus-induced membrane antigens: immunochemical characterization of Triton X-100 solubilized viral membrane antigens from EBV-superinfected Raji cells. Int J Cancer 23, 808–817.[CrossRef]
    [Google Scholar]
  28. Rickinson, A. B. & Kieff, E. ( 2007; ). Epstein–Barr virus. In Fields Virology, 5th edn, vol. 2, pp. 2655–2700. Edited by D. M. Knipe, P. M. Howley, D. E. Griffin, R. A. Lamb, M. A. Martin, B. Roizman & S. E. Straus. Philadelphia, PA: Lippincott Williams & Wilkins.
  29. Ross, L. J., Sanderson, M., Scott, S. D., Binns, M. M., Doel, T. & Milne, B. ( 1989; ). Nucleotide sequence and characterization of the Marek's disease virus homologue of glycoprotein B of herpes simplex virus. J Gen Virol 70, 1789–1804.[CrossRef]
    [Google Scholar]
  30. Silva, A. L., Omerovic, J., Jardetzky, T. S. & Longnecker, R. ( 2004; ). Mutational analyses of Epstein–Barr virus glycoprotein 42 reveal functional domains not involved in receptor binding but required for membrane fusion. J Virol 78, 5946–5956.[CrossRef]
    [Google Scholar]
  31. Spear, P. G. & Longnecker, R. ( 2003; ). Herpesvirus entry: an update. J Virol 77, 10179–10185.[CrossRef]
    [Google Scholar]
  32. Speck, P. & Longnecker, R. ( 2000; ). Infection of breast epithelial cells with Epstein-Barr virus via cell-to-cell contact. J Natl Cancer Inst 92, 1849–1851.[CrossRef]
    [Google Scholar]
  33. Strive, T., Borst, E., Messerle, M. & Radsak, K. ( 2002; ). Proteolytic processing of human cytomegalovirus glycoprotein B is dispensable for viral growth in culture. J Virol 76, 1252–1264.[CrossRef]
    [Google Scholar]
  34. Sullivan, D. C., Allen, G. P. & O'Callaghan, D. J. ( 1989; ). Synthesis and processing of equine herpesvirus type 1 glycoprotein 14. Virology 173, 638–646.[CrossRef]
    [Google Scholar]
  35. van Drunen Littel-van den Hurk, S. & Babiuk, L. A. ( 1986; ). Synthesis and processing of bovine herpesvirus 1 glycoproteins. J Virol 59, 401–410.
    [Google Scholar]
  36. Vey, M., Schafer, W., Reis, B., Ohuchi, R., Britt, W., Garten, W., Klenk, H. D. & Radsak, K. ( 1995; ). Proteolytic processing of human cytomegalovirus glycoprotein B (gpUL55) is mediated by the human endoprotease furin. Virology 206, 746–749.[CrossRef]
    [Google Scholar]
  37. Whealy, M. E., Robbins, A. K. & Enquist, L. W. ( 1990; ). The export pathway of the pseudorabies virus gB homolog gII involves oligomer formation in the endoplasmic reticulum and protease processing in the Golgi apparatus. J Virol 64, 1946–1955.
    [Google Scholar]
  38. Wolfer, U., Kruft, V., Sawitzky, D., Hampl, H., Wittmann-Liebold, B. & Habermehl, K. O. ( 1990; ). Processing of pseudorabies virus glycoprotein gII. J Virol 64, 3122–3125.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.007237-0
Loading
/content/journal/jgv/10.1099/vir.0.007237-0
Loading

Data & Media loading...

Most Cited This Month

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