1887

Journal of General Virology header logo

Volume 75, Issue 5

Role of -linked Oligosaccharide Chains in the Processing and Antigenicity of Measles Virus Haemagglutinin Protein Free

Abstract

The effects of -linked oligosaccharides on the haemagglutinin (H) protein of measles virus (MV) were assessed with respect to the processing and antigenicity of the molecule. The functional glycosylation sites on the H protein were determined by eliminating each of the five potential positions, Asn-168, Asn-187, Asn-200, Asn-215 and Asn-238, for -linked glycosylation by oligonucleotide-directed mutagenesis on a cDNA clone. Expression of the mutant H proteins in BHK-21 cells by a recombinant vaccinia virus encoding T7 polymerase indicated that the first four sites were used in the H glycoprotein for the addition of -linked oligosaccharide chains. Heterogeneity of oligosaccharide processing was demonstrated. One of the four glycosylation sites had a different carbohydrate structure from those of the other three glycosylation sites and this varied glycosylation was responsible for the appearance of two forms of the H protein. The functional glycosylation sites were systematically removed in various combinations from the H protein to form a panel of mutants in which the role of carbohydrate chains, singly or in different combinations, could be evaluated. Investigations of these glycosylation mutants indicated that (i) two of the four individual carbohydrate side-chains have a large influence on the antigenicity of the molecule; (ii) individual carbohydrate side-chains have little effect on the folding and oligomerization of the molecule, and are not sufficient or necessary alone to facilitate the transport of the molecule to the plasma membrane; (iii) at least two carbohydrate side-chains are required for the H protein to move along the exocytic pathway to the plasma membrane and various combinations of oligosaccharide side-chains, irrespective of the carbohydrate localizations, influence equally the processing of the molecule.

  • Received:
  • Accepted:
  • Published Online:
© The Journal of General Virology 1994
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-5-1043
1994-05-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/5/JV0750051043.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-5-1043&mimeType=html&fmt=ahah

References

  1. Alkhatib G., Briedis D. J. 1986; The predicted primary structure of the measles virus hemagglutinin. Virology 150:479–490
     [Google Scholar]
  2. Cattaneo R., 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]
  3. Copeland C. S., Doms R. W., Bolzau E. M., Webster R. G., Helenius A. 1986; Assembly of influenza hemagglutinin trimers and its role in intracellular transport. Journal of Cell Biology 103:1179–1191
     [Google Scholar]
  4. Copeland C. S., Zimmer K. P., Wagner K. R., Healey G. A., Mellman I., Helenius A. 1988; Folding, trimerization, and transport are sequential events in the biogenesis of influenza virus hemagglutinin. Cell 53:197–209
     [Google Scholar]
  5. Doms R. W., Ruusala A., Machamer C. M., Helenius A., Rose J. K. 1988; Differential effects of mutations in three domains on folding, quaternary structure, and intracellular transport of vesicular stomatitis virus G protein. Journal of Cell Biology 107:89–99
     [Google Scholar]
  6. Eschle D. 1988 Cloning and reconstruction of the entire measles virus genome Diploma thesis University of Zurich:
     [Google Scholar]
  7. Fuerst T. R., Niles E. G., Studier F. W., Moss B. 1986; Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proceedings of the National Academy of Sciences U.S.A: 838122–8126
     [Google Scholar]
  8. Gallagher P. J., Henneberry J. M., Sambrook J., Gething M. J. 1992; Glycosylation requirements for intracellular transport and function of the hemagglutinin of influenza virus. Journal of Virology 66:7136–7145
     [Google Scholar]
  9. Gething M. J., Mclammon K., Sambrook J. 1986; Expression of wild type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell 46:939–950
     [Google Scholar]
  10. Graves M. C. 1981; Measles virus polypeptides in infected cells studied by immune precipitation and one-dimensional peptide mapping. Journal of Virology 38:224–230
     [Google Scholar]
  11. Grigera P. R., Mathieu M. E., Wagner R. R. 1991; Effect of glycosylation on the conformational epitopes of the glycoprotein of vesicular stomatitis virus (New Jersey serotype). Virology 180:1–9
     [Google Scholar]
  12. Guan J. L., Cao H., Rose J. K. 1988; Cell-surface expression of a membrane-anchored form of the human chorionic gonadotropin αsubunit. Journal of Biological Chemistry 263:5306–5313
     [Google Scholar]
  13. Hannink M., Donoghue D. J. 1986; Cell surface expression of membrane-anchored V-sis gene products: glycosylation is not required for cell surface transport. Journal of Cell Biology 103:2311–2322
     [Google Scholar]
  14. Hardwick J. M., Hussell R. H. 1978; Glycoproteins of measles virus under reducing and nonreducing conditions. Journal of Virology 25:687–692
     [Google Scholar]
  15. Hu A., Sheshberadaran H., Norrby E., KöVamees J. 1993; Molecular characterization of epitopes on the measles virus hemagglutinin protein. Virology 192:351–354
     [Google Scholar]
  16. Hurtley S. M., Bole D, Hoover-Litty H., Helenius A., Copeland C. S. 1989; Interactions of misfolded influenza virus hemagglutinin with binding protein (Bip). Journal of Cell Biology 168:2117–2126
     [Google Scholar]
  17. Kornfeld R., Kornfeld S. 1985; Assembly of asparagine-linked oligosaccharides. Annual Review of Biochemistry 54:631–664
     [Google Scholar]
  18. Kreis T. E., Lodish H. F. 1986; Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface. Cell 46:929–937
     [Google Scholar]
  19. Machamer C. E., Florkiewicz R. Z., Rose J. K. 1985; A single V-linked oligosaccharide at either of the two normal sites is sufficient for transport of vesicular stomatitis virus G protein to the cell surface. Molecular and Cellular Biology 5:3074–3083
     [Google Scholar]
  20. Machamer C. E., Rose J. K. 1988; Vesicular stomatitis virus G proteins with altered glycosylation sites display temperature-sensitive intracellular transport and are subject to aberrant intermolecular disulfide bonding. Journal of Biological Chemistry 263:5955–5960
     [Google Scholar]
  21. Mackett M., Smith G. L., Moss B. 1985; The construction and characterization of vaccinia virus recombinants expressing foreign genes. In DNA Cloning 2 pp 191–211 Rickwood D., Hames B. D. Edited by Washington,D.C.: IRL Press;
     [Google Scholar]
  22. Matzuk M. M., Boime I. 1988; The role of the asparagine-linked oligosaccharides of the a subunit on the secretion and assembly of human chorionic gonadotropin. Journal of Cell Biology 106:1049–1059
     [Google Scholar]
  23. Mottet G., Portner A., Roux L. 1986; Drastic immunoreactivity changes between the immature and mature forms of the Sendai virus HN and F0 glycoprotein. Journal of Virology 59:132–141
     [Google Scholar]
  24. Ng D. T. W., Randall R. E., Lamb R. A. 1989; Intracellular maturation and transport of the SV5 type II glycoprotein hemagglutinin-neuraminidase: specific and transient association with GRP78-Bip in the endoplasmic reticulum and extensive internalization from the cell surface. Journal of Cell Biology 109:3273–3289
     [Google Scholar]
  25. Ng D. T. W., Hiebert S. W., Lamb R. A. 1990; Different roles of individual N-linked oligosaccharide chains in folding, assembly, and transport of the simian virus 5 hemagglutinin-neuraminidase. Molecular and Cellular Biology 10:1989–2001
     [Google Scholar]
  26. Norrby E., Oxman M. 1990; Measles virus. In Virology, 2nd edn.. pp 1013–1044 Fields B. N., Knipe D. M. Edited by New York: Raven Press;
     [Google Scholar]
  27. Norrby E., Chen S. N., Togashi T., Sheshberadaran H., Johnson K. P. 1982; Five measles virus antigens demonstrated by use of mouse hybridoma antibodies in productively infected tissue cells. Archives of Virology 71:1–11
     [Google Scholar]
  28. 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]
  29. Olden K., Parent J. B., White S. L. 1982; Carbohydrate moieties of glycoproteins. A re-evaluation of their function. Biochimica et biophysica acta 650:209–232
     [Google Scholar]
  30. Rademacher T. W., Parekh R. B., Dwek R. A. 1988; Glyco-biology. Annual Review of Biochemistry 57:785–838
     [Google Scholar]
  31. Sambrook J., Fritsch E. F., Maniatis T. 1989; . Molecular Cloning: A Laboratory Manual, 2nd edn.. New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  32. Sheshberadaran H., Chen S. H., Norrby E. 1983; Monoclonal antibodies against five structural components of measles virus. I. Characterization of antigenic determinants on nine strains of measles virus. Virology 128:341–353
     [Google Scholar]
  33. Sheshberadaran H., Norrby E. 1986; Characterization of epitopes on the measles virus hemagglutinin. Virology 152:58–65
     [Google Scholar]
  34. Sodora D. L., Cohen G. H., Eisenberg R. J. 1989; Influence of asparagine-linked oligosaccharide on antigenicity, processing, and cell surface expression of herpes simplex virus type I glycoprotein D. Journal of Virology 63:5184–5193
     [Google Scholar]
  35. Taylor A. K., Wall R. 1988; Selective removal of alpha heavy-chain glycosylation sites causes immunoglobulin A degradation and reduced secretion. Molecular and Cellular Biology 8:4197–4203
     [Google Scholar]
  36. Tikoo S. K., Parker M. D., Vandenhurk J. V., Kowalski J., Zamb T. J., Babiuk L. A. 1993; Role of N-linked glycans in antigenicity, processing, and cell surface expression of bovine herpesvirus 1 glycoprotein gIV. Journal of Virology 67:726–733
     [Google Scholar]
  37. Vidal S., Mottet G., Kolakofsky D., Roux L. 1989; Addition of high-mannose sugars must precede disulfide bond formation for proper folding of Sendai virus glycoproteins. Journal of Virology 63:892–900
     [Google Scholar]
  38. Waxham M. N., Merz D. C., Wolinsky J. S. 1986; Intracellular maturation of mumps virus hemagglutinin-neuraminidase glycoprotein: conformational changes detected with monoclonal antibodies. Journal of Virology 59:392–400
     [Google Scholar]
  39. William M. A., Lamb R. A. 1986; Determination of the orientation of an integral membrane protein and sites of glyco-sylation by oligonucleotide-directed mutagenesis: influenza B virus NB glycoprotein lacks a cleavable signal sequence and has an extracellular NH2-terminal region. Molecular and Cellular Biology 6:4317–4328
     [Google Scholar]
  40. Wright K. E., Salvato M. S., Buchmeier M. J. 1989; Neutralization epitopes of lymphocytic choriomeningitis virus are conformational and require both glycosylation and disulfide bonds for expression. Virology 171:417–426
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-5-1043
Loading
Role of N-linked Oligosaccharide Chains in the Processing and Antigenicity of Measles Virus Haemagglutinin Protein
J. Gen. Virol. 75, 1043 (1994); https://doi.org/10.1099/0022-1317-75-5-1043
/content/journal/jgv/10.1099/0022-1317-75-5-1043
/content/journal/jgv/10.1099/0022-1317-75-5-1043
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