1887

Journal of General Virology header logo

Volume 66, Issue 3

Respiratory Syncytial Virus Polypeptides. IV. The Oligosaccharides of the Glycoproteins Free

Abstract

SUMMARY

The cell-associated glycoproteins of respiratory syncytial (RS) virus included GP1 (90K), VP70 (70K), VGP48 (48K) and GP26 (26K). Although present in infected cells, there was no VP70 in purified virus. Trypsin treatment of infected cells removed 80 to 90% of VP70 as well as its products VGP48 and GP26. This suggested that most of the VP70 in the cell is located on the plasma membrane. The glycoproteins of purified RS virus (GP1, VGP48 and GP26) contain mannose, galactose and fucose as well as glucosamine, but the quantity of mannose in GP1 is low when compared to that of the other three sugars. The effects that follow the treatment of infected cells with the glycosylation inhibitors tunicamycin and monensin, and the treatment of the immunoprecipitated product of pulse-chase experiments with endonuclease H demonstrated that VP70 and its products contained -linked oligosaccharides, and that the oligosaccharides of the mature VGP48 subunit were of the complex type, while GP1 contained both - and -linked oligosaccharides. The non-glycosylated forms of VP70 and GP1 have estimated mol. wt. of 50K and 33K respectively. Therefore, the carbohydrate contribution to the mol. wt. of VP70 and GP1, as determined by PAGE, was equivalent to 20K for the former and 57K for the latter. The majority of the GP1 oligosaccharides were -linked, a form of sugar linkage not previously found among paramyxoviruses.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): cell-associated glycoproteins , envelope glycoproteins , glycoprotein oligosaccharides and RSV
© Journal of General Virology 1985
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-66-3-417
1985-03-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/66/3/JV0660030417.html?itemId=/content/journal/jgv/10.1099/0022-1317-66-3-417&mimeType=html&fmt=ahah

References

  1. Alonso F. V., Compans R. W. 1981; Differential effect of monensin on enveloped viruses that form at distinct plasma membrane domains. Journal of Cell Biology 89:700–705
     [Google Scholar]
  2. Arakawa M., Muramatsu T. 1974; Endo-β-N-acetylglucosaminidases acting on the carbohydrate moieties of glycoproteins: the differential specification of the enzymes from Streptomyces griseus and Diplococcus pneumoniae. Journal of Biochemistry 76:307–317
     [Google Scholar]
  3. Bernstein J. M., Hruska J. F. 1981; Respiratory syncytial virus proteins: identification by immunoprecipitation. Journal of Virology 38:278–285
     [Google Scholar]
  4. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
     [Google Scholar]
  5. Bretz R., Bretz H., Palade G. E. 1980; Distribution of terminal glycosyltransferases in hepatic Golgi fractions. Journal of Cell Biology 84:87–101
     [Google Scholar]
  6. Cash P., Pringle C. R., Preston C. M. 1979; The polypeptides of respiratory syncytial virus: products of cell-free protein synthesis and post-translational modifications. Virology 92:375–384
     [Google Scholar]
  7. Chatterjee S., Bradac J. A., Hunter E. 1982; Effect of monensin on Mason-Pfizer monkey virus glycoprotein synthesis. Journal of Virology 44:1003–1012
     [Google Scholar]
  8. Collins P. L., Huang Y. T., Wertz G. W. 1984; Identification of a tenth mRNA of respiratory syncytial virus and assignment of polypeptides to the 10 viral genes. Journal of Virology 49:572–578
     [Google Scholar]
  9. Diggelmann H. 1979; Biosynthesis of an unglycosylated envelope glycoprotein of Rous sarcoma virus in the presence of tunicamycin. Journal of Virology 30:799–804
     [Google Scholar]
  10. Dubovi E. J. 1982; Analysis of proteins synthesized in respiratory syncytial virus-infected cells. Journal of Virology 42:372–378
     [Google Scholar]
  11. Fernie B. F., Gerin J. L. 1982; Immunochemical identification of viral and nonviral proteins of the respiratory syncytial virus virion. Infection and Immunity 37:243–249
     [Google Scholar]
  12. Graves M. C. 1981; Measles virus polypeptides in infected cells studied by immune precipitation and onedimensional peptide mapping. Journal of Virology 38:224–230
     [Google Scholar]
  13. Gruber C., Levine S. 1983; Respiratory syncytial virus polypeptides. III. The envelope-associated proteins. Journal of General Virology 64:825–832
     [Google Scholar]
  14. Hanover J. A., Lennarz W. J., Young J. D. 1980; Synthesis of N- and O- linked glycopeptides in oviduct membrane preparations. Journal of Biological Chemistry 255:6713–6716
     [Google Scholar]
  15. Hardwick I. M., Bussell R. H. 1978; Glycoproteins of measles virus under reducing and nonreducing conditions. Journal of Virology 25:687–692
     [Google Scholar]
  16. Herp A., Wu A. M., Moschera J. 1979; Current concepts of the structure and nature of mammalian salivary mucous glycoproteins. Molecular and Cellular Biochemistry 23:27–41
     [Google Scholar]
  17. Herrler G., Compans R. W. 1983; Post-translational modification and intracellular transport of mumps virus glycoproteins. Journal of Virology 47:354–362
     [Google Scholar]
  18. Holmes K. V., Doller E. W., Sturman L. S. 1981; Tunicamycin resistant glycosylation of a coronavirus glycoprotein: demonstration of a novel type of viral glycoprotein. Virology 115:334–344
     [Google Scholar]
  19. Huang Y. T., Wertz G. W. 1983; Respiratory syncytial virus mRNA coding assignments. Journal of Virology 46:667–672
     [Google Scholar]
  20. Johnson D. C., Schlesinger M. J. 1980; Vesicular stomatitis virus and Sindbis virus glycoprotein transport to the cell surface is inhibited by ionophores. Virology 103:407–424
     [Google Scholar]
  21. Johnson D. C., Spear P. G. 1982; Monensin inhibits the processing of herpes simplex virus glycoproteins, their transport to the cell surface, and the egress of virions from infected cells. Journal of Virology 43:1102–1112
     [Google Scholar]
  22. Johnson D. C., Spear P. G. 1983; O-linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus. Cell 32:987–997
     [Google Scholar]
  23. Kohama T., Shimizu K., Ishida N. 1978; Carbohydrate composition of the envelope glycoproteins of Sendai virus. Virology 90:226–234
     [Google Scholar]
  24. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 221:680–685
     [Google Scholar]
  25. Leavitt R., Schlesinger S., Kornfeld S. 1977; Tunicamycin inhibits glycosylation and multiplication of Sindbis and vesicular stomatitis viruses. Journal of Virology 21:375–385
     [Google Scholar]
  26. Lee G. T.-Y., Para M. F., Spear P. G. 1982; Location of the structural genes for glycoproteins gD and gE and for other polypeptides in the S component of herpes simplex virus type 1 DNA. Journal of Virology 43:41–49
     [Google Scholar]
  27. Lehle L., Tanner W. 1976; The specific site of tunicamycin inhibition in the formation of dolichol-bound N- acetylglucosamine derivatives. FEBS Letters 71:167–170
     [Google Scholar]
  28. Levine S. 1977; Polypeptides of respiratory syncytial virus. Journal of Virology 21:427–431
     [Google Scholar]
  29. Levine S., Hamilton R. 1969; Kinetics of the respiratory syncytial virus growth cycle in HeLa cells. Archiv für die gesamte Virusforschung 28:122–132
     [Google Scholar]
  30. Levine S., Buthala D. A., Hamilton R. D. 1971; Late stage synchronization of respiratory syncytial virus replication. Virology 45:390–400
     [Google Scholar]
  31. Morrison T. G., Simpson D. 1980; Synthesis, stability and cleavage of Newcastle disease virus glycoproteins in the absence of glycosylation. Journal of Virology 36:171–180
     [Google Scholar]
  32. Nakamura K., Compans R. W. 1978; Effects of glucosamine, 2-deoxyglucose and tunicamycin on glycosylation, sulfation, and assembly of influenza viral proteins. Virology 84:303–319
     [Google Scholar]
  33. Niemann H., Klenk H.-D. 1981; Coronavirus glycoprotein E1, a new type of viral glycoprotein. Journal of Molecular Biology 153:993–1010
     [Google Scholar]
  34. Niemann H., Boschek B., Evans D., Rosing M., Tamura T., Klenk H.-D. 1982; Post-translational glycosylation of coronavirus glycoprotein E-1: inhibition by monensin. EMBO Journal 1:1499–1504
     [Google Scholar]
  35. Oker-Blom C., Kalkkinen N., KÄÄriÄinen L., Pettersson R. F. 1983; Rubella virus contains one capsid protein and three envelope glycoproteins, E1, E2a, and E2b. Journal of Virology 46:964–973
     [Google Scholar]
  36. Payne L. G., Kristensson K. 1982; The effect of cytochalasin D and monensin on enveloped vaccinia virus release. Archives of Virology 74:11–20
     [Google Scholar]
  37. Peeples M., Levine S. 1979; Respiratory syncytial virus polypeptides: their location in the virion. Virology 95:137–145
     [Google Scholar]
  38. Peeples M., Levine S. 1980; Metabolic requirements for the maturation of respiratory syncytial virus. Journal of General Virology 50:81–88
     [Google Scholar]
  39. Pesonen M., Kääriäinen L. 1982; Incomplete complex oligosaccharides in Semliki Forest virus envelope proteins arrested within the cell in the presence of monensin. Journal of Molecular Biology 158:213–230
     [Google Scholar]
  40. Pressman B. C. 1976; Biological application of ionophores. Annual Review of Biochemistry 45:501–530
     [Google Scholar]
  41. Schachter H. 1974; The subcellular sites of glycosylation. Biochemical Society Symposia 40:57–71
     [Google Scholar]
  42. Scheid A., Choppin P. W. 1977; Two disulfide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology 80:54–66
     [Google Scholar]
  43. Schwalbe J. C., Hightower L. E. 1982; Maturation of the envelope glycoproteins of Newcastle disease virus on cellular membranes. Journal of Virology 41:947–957
     [Google Scholar]
  44. Schwarz R. T., Rohrschneider J. M., Schmidt M. F. G. 1976; Suppression of glycoprotein formation of Semliki Forest, influenza and avian sarcoma virus by tunicamycin. Journal of Virology 19:782–791
     [Google Scholar]
  45. Shida H., Dales S. 1981; Biogenesis of vaccinia: carbohydrate of the hemagglutinin molecule. Virology 111:56–72
     [Google Scholar]
  46. Slomiany A., Slomiany B. L. 1978; Structures of the acidic oligosaccharides isolated from rat sublingual glycoprotein. Journal of Biological Chemistry 253:7301–7306
     [Google Scholar]
  47. Stallcup K. C., Fields B. N. 1981; The replication of measles virus in the presence of tunicamycin. Virology 108:391–404
     [Google Scholar]
  48. Strous G. J. A. M., Lodish H. F. 1980; Intracellular transport of secretory and membrane proteins in hepatoma cells infected by vesicular stomatitis virus. Cell 22:709–717
     [Google Scholar]
  49. Tabas I., Kornfeld S. 1979; Purification and characterization of a rat liver Golgi α-mannosidase capable of processing asparagine-linked oligosaccharides. Journal of Biological Chemistry 254:11655–11663
     [Google Scholar]
  50. Takatsuki A., Arima K., Tamura G. 1971; Tunicamycin, a new antibiotic. I. Isolation and characterization of tunicamycin. Journal of Antibiotics 24:215–223
     [Google Scholar]
  51. Tarentino A. L., Maley F. 1974; Purification and properties of an endo-β-N-acetylglucosaminidase from Slreptomyces griseus. Journal of Biological Chemistry 249:811–817
     [Google Scholar]
  52. 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]
  53. Thomas D. B., Winzler R. J. 1969; Structural studies on human erythrocyte glycoproteins: alkali-labile oligosaccharides. Journal of Biological Chemistry 244:5943–5946
     [Google Scholar]
  54. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
     [Google Scholar]
  55. Tulsiani D. R. P., Hubbard S. C., Robbins P. W., Touster O. 1982; α-d-mannosidases of rat liver Golgi membranes. Journal of Biological Chemistry 257:3660–3668
     [Google Scholar]
  56. Vaessen R. T. M. J., Kreike J., Groot G. S. P. 1981; Protein transfer to nitrocellulose filters. FEBS Letters 124:193–196
     [Google Scholar]
  57. Waechter C. J., Lennarz W. J. 1976; The role of polyprenol-linked sugars in glycoprotein synthesis. Annual Review of Biochemistry 45:95–111
     [Google Scholar]
  58. Walsh E. E., Hruska J. 1983; Monoclonal antibodies to respiratory syncytial virus proteins. Identification of the fusion protein. Journal of Virology 47:171–177
     [Google Scholar]
  59. Wenske E. A., Courtney R. J. 1983; Glycosylation of herpes simplex virus type 1 gC in the presence of tunicamycin. Journal of Virology 46:297–301
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-66-3-417
Loading
Respiratory Syncytial Virus Polypeptides. IV. The Oligosaccharides of the Glycoproteins
J. Gen. Virol. 66, 417 (1985); https://doi.org/10.1099/0022-1317-66-3-417
/content/journal/jgv/10.1099/0022-1317-66-3-417
/content/journal/jgv/10.1099/0022-1317-66-3-417
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