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. 2004 Feb 2;164(2):458–466. doi: 10.1016/0042-6822(88)90560-0

Role of oligosaccharides in the structure and function of respiratory syncytial virus glycoproteins

Dennis M Lambert 1
PMCID: PMC7130872  PMID: 3369089

Abstract

The contribution of oligosaccharides to the structural and functional make-up of respiratory syncytial (RS) virus G and F proteins was investigated by observing the effects of various oligosaccharide-specific enzymes on their molecular size as well as on virus infectivity. The N-linked oligosaccharides of the F protein were completely removed by endoglycosidase F and N-glycanase. Addition of oligosaccharides to F protein during synthesis was completely partially resistant to TM resulting in an 80-kDa form designated GTM. The G protein was estimated to contain approximately 3% N-linked and 55% O-linked carbohydrates, based on migration of G and GTM in polyacrylamide gels. Furthermore, treatment of detergent-extracted G protein with endoglycosidase F and endo-α-N-acetylgalactosaminidase, enzymes that specifically cleave N-linked and O-linked oligosaccharides, respectively, generated a variety of partially unglycosylated species, ranging in molecular weight from approximately 80 to 40 kDa. Virus infectivity was sensitive to limited removal of N-linked or O-linked oligosaccharides by endoglycosidases under conditions which did not greatly alter the molecular weight of the G protein. Thus, G and F protein oligosaccharides readily accessible to enzymatic removal are presumed to play an important role in the infectious process.

Reference

  1. Collins P.L., Huang Y.T., Wertz G.W. Identification of a tenth mRNA of respiratory syncytial virus and assignment of polypeptides to the 10 viral genes. J. Virol. 1984;49:572–578. doi: 10.1128/jvi.49.2.572-578.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dore-Duffy P., Howe C. Vol. 157. 1978. N-Acetylneuraminic acid (NANA) in measles virus; pp. 622–625. (Proc. Soc. Exp. Biol. Med.). [DOI] [PubMed] [Google Scholar]
  3. Dubovi E.J. Analysis of proteins synthesized in respiratory syncytial virus-infected cells. J. Virol. 1982;42:372–378. doi: 10.1128/jvi.42.2.372-378.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fernie B., Dapolito G., Cote P.J., Gerin J.L. Kinetics of respiratory syncytial virus glycoproteins. J. Gen. Virol. 1985;66:1983–1990. doi: 10.1099/0022-1317-66-9-1983. [DOI] [PubMed] [Google Scholar]
  5. Fernie B., Gerin J.L. Immunochemical identification of viral and non-viral proteins of the respiratory syncytial virus virion. Infect. Immun. 1982;37:243–249. doi: 10.1128/iai.37.1.243-249.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gibson R., Leavitt R., Kornfeld S., Schlesinger S. Synthesis and infectivity of vesicular stomatitis virus containing nonglycosylated G protein. Cell. 1978;13:671–679. doi: 10.1016/0092-8674(78)90217-9. [DOI] [PubMed] [Google Scholar]
  7. Griffiths G., Quinn P., Warren G. 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. J. Cell Biol. 1983;96:835–850. doi: 10.1083/jcb.96.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gruber C., Levine S. Respiratory syncytial virus polypeptides. III. The envelope-associated proteins. J. Gen. Virol. 1983;64:825–832. doi: 10.1099/0022-1317-64-4-825. [DOI] [PubMed] [Google Scholar]
  9. Gruber C., Levine S. Respiratory syncytial virus polypeptides. IV. The oligosaccharides of the glycoproteins. J. Gen. Virol. 1985;66:417–432. doi: 10.1099/0022-1317-66-3-417. [DOI] [PubMed] [Google Scholar]
  10. Holmes K.V., Doller E.W., Sturman L.S. Tunicamycin resistant glycosylation of a coronavirus glycoprotein: Demonstration of a novel type of viral glycoprotein. Virology. 1981;115:334–344. doi: 10.1016/0042-6822(81)90115-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang Y.T., Collins P.L., Wertz G.W. Characterization of the 10 proteins of human respiratory syncytial virus: Identification of a fourth envelope-associated protein. Virus Res. 1985;2:157–173. doi: 10.1016/0168-1702(85)90246-1. [DOI] [PubMed] [Google Scholar]
  12. Johnson D.C., Spear P.G. O-Linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus. Cell. 1983;32:987–997. doi: 10.1016/0092-8674(83)90083-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kawaoka Y., Naeve C.W., Webster R.G. Is virulence of H5N2 influenza viruses in chickens associated with loss of carbohydrate from the hemagglutinin? Virology. 1984;139:303–316. doi: 10.1016/0042-6822(84)90376-3. [DOI] [PubMed] [Google Scholar]
  14. Kingsbury D.W., Bratt M.A., Choppin P.W., Hanson R.P., Hosaka V., ter Meulen V., Norrby E., Plowright W., Rott R., Wunner W.H. Paramyxoviridae. Intervirology. 1978;10:137. [Google Scholar]
  15. Laemmli U.K. Cleavage of structural proteins during the assembly of bacteriophage T4. Nature (London) 1970;227:680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lambert D.M., Pons M.W. Respiratory syncytial virus glycoproteins. Virology. 1983;130:204–214. doi: 10.1016/0042-6822(83)90128-9. [DOI] [PubMed] [Google Scholar]
  17. Lambert D.M., Pons M.W. Characterization of the glycoproteins of respiratory syncytial virus. In: Compans R., Bishop D., editors. Nonsegmented Negative Strand Viruses. Academic Press; New York: 1984. pp. 369–376. [Google Scholar]
  18. Lambert D.M., Pons M.W., Mbuy G.N., Dorsch-Hasler K. Nucleic acids of respiratory syncytial virus. J. Virol. 1980;36:837–846. doi: 10.1128/jvi.36.3.837-846.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Laskey R.A., Mills A.D. Quantitative film detection of3H and14C in polyacrylamide gels by fluorography. Eur. J. Biochem. 1975;56:335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  20. Leavitt R., Schlesinger S., Kornfeld S. Tunicamycin inhibits glycosylation and multiplication of Sindbis and vesicular stomatitis virus. J. Virol. 1977;21:375–385. doi: 10.1128/jvi.21.1.375-385.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Levine S. Polypeptides of respiratory syncytial virus. J. Virol. 1977;21:427–431. doi: 10.1128/jvi.21.1.427-431.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Levine S., Klaiber-Franco R., Paradiso P.R. Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. J. Gen. Virol. 1987;68:2521–2524. doi: 10.1099/0022-1317-68-9-2521. [DOI] [PubMed] [Google Scholar]
  23. Nakamura K., Homma M., Compans R.W. Effect of tunicamycin on the replication of Sendai virus. Virology. 1982;119:474–487. doi: 10.1016/0042-6822(82)90106-4. [DOI] [PubMed] [Google Scholar]
  24. Niemann H., Boschek B., Evans D., Rosing M., Tamura T., Klenk H.-D. Post-translational glycosylation of corona-virus glycoprotein E1: Inhibition by monensin. EMBO J. 1982;1:1499–1504. doi: 10.1002/j.1460-2075.1982.tb01346.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Niemann H., Geyer R., Klenk H.-D., Linder D., Stirm S., Wirth M. The carbohydrates of mouse hepatitis virus (MHV) A59: Structures of the O-glycosidically linked oligosaccharides of glycoprotein E1. EMBO J. 1984;3:665–670. doi: 10.1002/j.1460-2075.1984.tb01864.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Olofsson S., Blomberg J., Lycke E. O-Glycosidic carbohydrate-peptide linkages of herpes simplex virus glycoproteins. Arch. Virol. 1981;70:321–329. doi: 10.1007/BF01320247. [DOI] [PubMed] [Google Scholar]
  27. Peeples M., Levine S. Respiratory syncytial virus polypeptides: Their location in the virion. Virology. 1979;95:137–145. doi: 10.1016/0042-6822(79)90408-2. [DOI] [PubMed] [Google Scholar]
  28. Pringle C.R., Shirodaria P.V., Gimenez H.B., Levine S. Antigen and polypeptide synthesis by temperature-sensitive mutants of respiratory syncytial virus. J. Gen. Virol. 1981;54:173–183. doi: 10.1099/0022-1317-54-1-173. [DOI] [PubMed] [Google Scholar]
  29. Richman A.V., Pedreira F.A., Tauraso N.M. Attempts to demonstrate hemagglutination and hemdsorption by respiratory syncytial virus. Appl. Microbiol. 1971;21:1099. doi: 10.1128/am.21.6.1099-1100.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Satake M., Coligan J.E., Elango N., Norrby E., Venkatesan S. Respiratory syncytial virus envelope glycoprotein (G) has a novel structure. Nucleic Acids Res. 1985;13:7795–7812. doi: 10.1093/nar/13.21.7795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sharon N., Lis H. Glycoproteins: Research booming on long-ignored, ubiquitous compounds. Chem. Engineering News. 1981;59:21–44. doi: 10.1007/BF00238511. [DOI] [PubMed] [Google Scholar]
  32. Shida H., Dales S. Biogenesis of vaccinia: Carbohydrate of the hemagglutinin molecule. Virology. 1981;111:56–72. doi: 10.1016/0042-6822(81)90653-x. [DOI] [PubMed] [Google Scholar]
  33. Sjoblom I., Lundstrom M., Sjogren-Jansson E., Glorioso J.C., Jeansson S., Olofsson S. Demonstration and mapping of highly carbohydrate-dependent epitopes in the herpes simplex virus type 1-specified glycoprotein C. J. Gen. Virol. 1987;68:545–554. doi: 10.1099/0022-1317-68-2-545. [DOI] [PubMed] [Google Scholar]
  34. Takatsuki A., Arima K., Tamura G. Tunicamycin, a new antibiotic. I. Isolation and characterization of tunicamycin. J. Antibiot. 1971;24:224–231. doi: 10.7164/antibiotics.24.215. [DOI] [PubMed] [Google Scholar]
  35. Takatsuki A., Kono K., Tamura G. Inhibition of biosynthesis of polyisoprenol sugars in chick embryo microsomes by tunicamycin. Agric. Biol. Chem. 1975;39:2089–2091. [Google Scholar]
  36. Towbin H., Staehelin T., Gordon J. Vol. 76. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedures and some applications; pp. 4350–4354. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Umemoto J., Bhavanandan V.P., Davidson E.A. Purification and properties of an endo-α- N-acetyl-d-galactosaminidase from Diplococcus pneumoniae. J. Biol. Chem. 1977;252:8609–8614. [PubMed] [Google Scholar]
  38. Walsh E.E., Hruska J. Monoclonal antibodies to respiratory syncytial virus proteins: Identification of the fusion protein. J. Virol. 1983;47:171–177. doi: 10.1128/jvi.47.1.171-177.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Walsh E.E., Schlesinger J.J., Brandiss M.W. Purification and characterization of GP90, on of the envelope glycoproteins of respiratory syncytial virus. J. Gen. Virol. 1984;65:761–767. doi: 10.1099/0022-1317-65-4-761. [DOI] [PubMed] [Google Scholar]
  40. Wertz G.W., Collins P.L., Huang Y., Gruber C., Levine S., Ball A.L. Vol. 82. 1985. Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein; pp. 4075–4079. (Proc. Natl. Acad. Sci. USA). [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wunner W.H., Pringle C.R. Respiratory syncytial virus proteins. Virology. 1976;73:228–243. doi: 10.1016/0042-6822(76)90077-5. [DOI] [PubMed] [Google Scholar]

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