1887

Abstract

The measles virus (MV) haemagglutinin (HA) is a class 2 glycoprotein by means of which the virus particle attaches to the host cell receptor. We have previously expressed this glycoprotein as a vaccinia recombinant virus and have shown that the HA glycoprotein synthesized is indistinguishable from that coded by MV. In the present study, we report that in RK13 cells a soluble form (sHA) of the HA is secreted into the medium. We show by SDS-PAGE and sucrose density gradient centrifugation that the sHA is a dimer and is smaller than the cell-associated form. Using a variety of inhibitors the production of sHA was shown to be a late event, probably occurring at the membrane; only fully glycosylated molecules were found in sHA. Finally, we demonstrate that sHA retains its antigenicity with conformation-dependent MAbs and its receptor recognition function. We conclude that sHA is a valuable tool for use in studies of the structure and function of the MV HA glycoprotein.

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1994-12-01
2024-12-13
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References

  1. Cattaneo G., Rose J. K. 1993; Cell fusion by the envelope glycoprotein of persistent measles virus which caused lethal human brain disease. Journal of Virology 67:1493–1502
    [Google Scholar]
  2. Doms R. W., Russ G., Yewdell J. W. 1989; Brefeldin A redistributes resident and itinerant Golgi proteins to the endoplasmic reticulum. Journal of Cell Biology 109:61–72
    [Google Scholar]
  3. Dörig R. E., Marcil A., Chopra A., Richardson C. D. 1993; The human CD46 molecule is a receptor for measles virus (Edmonston Strain). Cell 75:295–305
    [Google Scholar]
  4. Drillien R., Spehner D., Kirn A., Giraudon P., Buckland R., Wild F., Lecocq J. P. 1988; Protection of mice from fatal encephalitis by vaccination with vaccinia virus recombinants encoding either the hemagglutinin or the fusion protein. Proceedings of the National Academy of Sciences, U.S.A 87:648–652
    [Google Scholar]
  5. Dunster L. M., Schneider-Schaulies J., Löffler S., Lankes W., Schwartz-Albiez R., Lottspeich F., ter Meulen V. 1994; Moesin: a cell membrane protein linked with susceptibility to measles virus infection. Virology 198:265–274
    [Google Scholar]
  6. Fenouillet E., Clerget-Raslain B., Gluckman J. C., Guftard D., Montagnier L., Bahraoui E. 1989; Role of N-linked glycans in the interaction between the envelope glycoprotein of human immunodeficiency virus and its CD4 cellular receptor. Journal of Experimental Medicine 169:807–822
    [Google Scholar]
  7. Gallagher P. J., Henneberry J. M., Sambrook J. F., Gething M.-J. H. 1992; Glycosylation requirements for intracellular transport and function of the hemagglutinin of influenza virus. Journal of Virology 66:7136–7145
    [Google Scholar]
  8. Giraudon P., Wild T. F. 1985; Correlation between epitopes on hemagglutinin of measles virus and biological activities: passive protection by monoclonal antibodies is related to their hemagglutination inhibiting activity. Virology 144:46–58
    [Google Scholar]
  9. Griffiths G., Quinn P., Warren G. 1983; Dissection of the Golgi complex. I. Monensin inhibits the transport of viral membrane proteins from medial to trans Golgi cisternae in baby hamster kidney cells infected with Semliki Forest virus. Journal of Cell Biology 96:835–850
    [Google Scholar]
  10. Hardwick J. M., Bussel R. H. 1978; Glycoproteins of measles virus under reducing and non reducing conditions. Journal of Virology 25:687–692
    [Google Scholar]
  11. Hu A., Cattaneo R., Schwartz S., Norrby E. 1994; Role of N-linked oligosaccharide chains in the processing and antigenicity of measles virus haemagglutinin protein. Journal of General Virology 75:1043–1052
    [Google Scholar]
  12. Kornfeld R., Kornfeld S. 1985; Assembly of asparagine-linked oligosaccharides. Annual Review of Biochemistry 54:631–664
    [Google Scholar]
  13. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  14. Li Y., Luo L., Rasool N., Yong Kang C. 1993; Glycosylation is necessary for the correct folding of human immunodeficiency virus gpl20 in CD4 binding. Journal of Virology 67:584–588
    [Google Scholar]
  15. Lippincott-Schwartz J., Yuan L. C., Bonifacino J. S., Klausner R. D. 1989; Rapid redistribution of Golgi proteins into the endoplasmic reticulum in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER. Cell 56:801–813
    [Google Scholar]
  16. Liszewski M. K., Post T. W., Atkinson J. P. 1991; Membrane cofactor protein (MCP or CD46): newest member of the regulators of complement activation gene cluster. Annual Review of Immunology 9:431–455
    [Google Scholar]
  17. Mcginnes L., Sergel T., Morrison T. 1993; Mutations in the transmembrane domain of the HN protein of newcastle disease virus affect the structure and activity of the protein. Virology 196:101–110
    [Google Scholar]
  18. Malvoisin E., Wild T. F. 1993; Measles virus glycoproteins: studies on the structure and interaction of the haemagglutinin and fusion proteins. Journal of General Virology 74:2365–2372
    [Google Scholar]
  19. Malvoisin E., Wild T. F. 1994; The role of N-glycosylation in cell fusion induced by a vaccinia recombinant virus expressing both measles virus glycoproteins. Virology 200:11–20
    [Google Scholar]
  20. Mirza A. M., Sheehan J. P., Hardy L. W., Glickman R. L., Iorio R. M. 1993; Structure and function of a membrane anchorless form of the hemagglutinin-neuraminidase glycoprotein of newcastle disease virus. Journal of Biological Chemistry 268:21425–21431
    [Google Scholar]
  21. Naniche D., Varior-Krishnan G., Cervoni F., Wild F., Rossi B., Rabourdin-Combe C., Gerlier D. 1993; Human membrane cofactor protein (CD46) acts as a cellular receptor for measles virus. Journal of Virology 67:6025–6032
    [Google Scholar]
  22. Ogura H., Sato H., Kamiya S., Nakamura S. 1991; Glycosylation of measles virus haemagglutinin protein in infected cells. Journal of General Virology 72:2679–2684
    [Google Scholar]
  23. Olden K., Bernard B. A., Humphries M. J., Yeo T. K., Yeo K. T., White S. L., Newton S. A., Bauer H. C., Parent J. B. 1985; Function of glycoprotein glycans. Trends in Biochemical Sciences 10:78–82
    [Google Scholar]
  24. Sampath D., Varki A., Freeze H. H. 1992; The spectrum of incomplete N-linked oligosaccharides synthesized by endothelial cells in the presence of brefeldin A. Journal of Biological Chemistry 267:4440–4455
    [Google Scholar]
  25. Sheshberadaran H., Norrby E. 1986; Characterization of epitopes on the measles virus hemagglutinin. Virology 152:58–65
    [Google Scholar]
  26. Sparrow R. L., Mckenzie I. F. C. 1983; HuLy-m5: a unique antigen physically associated with HLA molecules. Human Immunology 1:1–15
    [Google Scholar]
  27. Spies C. P., Compans R. W. 1993; Alternate pathways of secretion of simian immunodeficiency virus envelope glycoproteins. Virology 67:6535–6541
    [Google Scholar]
  28. Tartakoff A. M. 1983; Perturbation of vesicular traffic with the carboxylic acid ionophoremonensin. Cell 32:1026–1028
    [Google Scholar]
  29. Tartakoff A. M., Vassalli P. 1977; Plasma cell immunoglobulin secretion. Arrest is accompanied by alterations of the Golgi complex. Journal of Experimental Medicine 146:1332–1345
    [Google Scholar]
  30. Thompson S. D., Laver W. G., Murti K. G., Portner A. 1988; Isolation of a biologically active soluble form of the hemagglutinin-neuraminidase protein of Sendai virus. Journal of Virology 62:4653–4660
    [Google Scholar]
  31. Tifft C. J., Proia R. L., Camerini-Otero R. D. 1992; The folding and cell surface expression of CD4 require glycosylation. Journal of Biological Chemistry 267:3268–3273
    [Google Scholar]
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