1887

Abstract

The fusion (F) glycoprotein of measles virus, a structural component of the virion envelope, contains four potential sites for attachment of -linked oligosaccharides. Three are located in the F subunit of the protein and one in the signal peptide. Four mutants were constructed by oligonucleotide-directed mutagenesis, in each case changing one -linked glycosylation site from Asn-X-Ser/Thr to Ser-X-Ser/Thr. The wild-type and altered forms of the F protein were expressed in BHK-21 and HeLa T4 cells by use of the recombinant vaccinia virus-encoding T7 polymerase system. Analysis of these proteins revealed that three (residues 29, 61 and 67) potential sites for addition of -linked glycans in the F subunit are actually utilized. The functional glycosylation sites were systematically removed in all possible combinations from the F protein to form a panel of mutants from which the role of carbohydrates, singly or in various combinations, could be evaluated. One singlesite mutant protein lacking the glycosylation site of Asn-67 was processed, transported to the cell surface and could induce cell fusion. However, the other two single-site mutant proteins with deletions of glycosylation sites Asn-29 or Asn-61 exhibited a defect in processing, were not transported to cell surface and thus induced no cell fusion. The absence of any two of the three or of all three glycosylation sites resulted in protein retention in the endoplasmic reticulum. Therefore, it appears that glycosylation of sites Asn-29 and Asn-61 has important roles in maintaining the native structure of the F protein.

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1995-03-01
2021-10-28
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References

  1. Alkhatib G., Shen S.-H., Briedis D., Richardson C., Massie B., Weinberg R., Smith D., Taylor J., Paoletti E., Roder J. 1994; Functional analysis of N-linked glycosylation mutants of the measles virus fusion protein synthesized by recombinant vaccinia virus. Journal of Virology 68:1522–1531
    [Google Scholar]
  2. Barrett T., Clarke D. K., Evans S. A., Rima B. K. 1987; The nucleotide sequence of the gene encoding the F protein of canine distemper virus: a comparison of the deduced amino acid sequence with other paramyxoviruses. Virus Research 8:373–386
    [Google Scholar]
  3. Cattaneo R., Rose J. K. 1993; Cell fusion by the envelope glycoproteins of persistent measles virus which caused lethal human brain disease. Journal of Virology 67:1493–1502
    [Google Scholar]
  4. Döric 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]
  5. 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 USA 83:8122–8126
    [Google Scholar]
  6. Hu A., Cattaneo R., Schwartz S., Norrby E. 1994a; Role of A-linked oligosaccharide chains in the processing and antigenicity of measles virus haemagglutinin protein. Journal of General Virology 75:1043–1052
    [Google Scholar]
  7. Hu A., Kövamees J., Norrby E. 19946; Intracellular processing and antigenic maturation of the measles virus haemagglutinin protein. Archives of Virology 136:239–253
    [Google Scholar]
  8. Kövamees J., Blixenkrone-Möller M., Sharma B., Örvell C., Norrby E. 1991; The nucleotide sequence and deduced amino acid composition of the haemagglutinin and fusion proteins of the morbillivirus phocid distemper virus. Journal of General Virology 72:2959–2966
    [Google Scholar]
  9. 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]
  10. Malvoisin E., Wild T. F. 1994; The role of TV-glycosylation in cell fusion induced by a vaccinia recombinant virus expressing both measles virus glycoproteins. Virology 200:11–20
    [Google Scholar]
  11. Morrison T. G., Ward L. J., Semerjian A. 1985; Intracellular processing of Newcastle disease virus fusion glycoprotein. Journal of Virology 53:851–857
    [Google Scholar]
  12. Morrison T. G., Portner A. 1991; Structure, function, and intracellular processing of the glycoprotein of paramyxoviridae. In The Paramyxoviruses pp 347–382 Edited by Kingsbury D. W. New York: Plenum Press;
    [Google Scholar]
  13. 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]
  14. Naniche D., Carior-Krishana G., Cervoni F., Wild T. 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]
  15. Norrby E., Oxman M. 1990; Measles virus. In Virology 2nd end, pp 1013–1044 Edited by Fields B. Ν. and others New York: Raven Press;
    [Google Scholar]
  16. 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]
  17. Rademacher T. W., Parekh R. B., Dwek R. A. 1988; Glycobiology. Annual Review of Biochemistry 57:785–838
    [Google Scholar]
  18. Richardson C., Hull D., Greer H., Hassle K., Berkovich A., Englund G., Bellini W., Rima B., Lazzarini R. 1986; The nucleotide sequence of the mRNA encoding the fusion protein of measles virus (Edmonston strains): a comparison of fusion protein from several different paramyxoviruses. Virology 155:508–523
    [Google Scholar]
  19. Scheid A., Choppin P. W. 1974; Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis, and infectivity by proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology 57:470–490
    [Google Scholar]
  20. Scheid A., Choppin P. W. 1977; Two disulphide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology 80:54–66
    [Google Scholar]
  21. SpielhOfer P. 1990 Structure and expression of the measles virus envelope genes Diploma thesis University of Zurich; Zurich, Switzerland.:
    [Google Scholar]
  22. Tsukiyama K., Yoshikawa Y., Yamanouchi K. 1988; Fusion glycoprotein (F) of rinderpest virus: entire nucleotide sequence of the F mRNA, and several features of the F protein. Virology 164:523–530
    [Google Scholar]
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