1887

Abstract

Most street rabies virus G proteins have two -glycosylation sites, i.e. Asn and Asn, whereas additional sites are found in fixed (laboratory adapted) viruses. In this study, we performed a pseudotyped virus assay using G-deficient rabies virus and demonstrated that single--glycan additions to the G protein of street rabies virus strain 1088, which are found in adapted strains, enhanced virus production in neural and non-neural cell lines, while additions to Asn or Asn enhanced production greatly. Moreover, we found that -glycan additions at Asn or Asn facilitated the production of cell-associated virus. In contrast, deletion of the sequon at Asn reduced viral production, while a deletion at Asn resulted in extensive loss of production. Furthermore, G proteins lacking an -glycan at Asn failed to fold into their correct structure and lost their fusion activity, indicating that Asn -glycosylation is important for the functional expression of street virus G proteins.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.047852-0
2013-02-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/94/2/270.html?itemId=/content/journal/jgv/10.1099/vir.0.047852-0&mimeType=html&fmt=ahah

References

  1. Badrane H., Tordo N. 2001; Host switching in Lyssavirus history from the Chiroptera to the Carnivora orders. J Virol 75:8096–8104 [View Article][PubMed]
    [Google Scholar]
  2. Gaudin Y. 1997; Folding of rabies virus glycoprotein: epitope acquisition and interaction with endoplasmic reticulum chaperones. J Virol 71:3742–3750[PubMed]
    [Google Scholar]
  3. Hiramatsu K., Mifune K., Mannen K., Nishizono A., Kawano H., Ito Y., Kawai A. 1992; Mapping of the antigenic determinants recognized by monoclonal antibodies against the M2 protein of rabies virus. Virology 187:472–479 [View Article][PubMed]
    [Google Scholar]
  4. Irie T., Matsuda Y., Honda Y., Morimoto K., Kawai A. 2002; Studies on the escape mutants of rabies virus which are resistant to neutralization by a highly conserved conformational epitope-specific monoclonal antibody #1-46-12. Microbiol Immunol 46:449–461[PubMed] [CrossRef]
    [Google Scholar]
  5. Ito N., Takayama M., Yamada K., Sugiyama M., Minamoto N. 2001; Rescue of rabies virus from cloned cDNA and identification of the pathogenicity-related gene: glycoprotein gene is associated with virulence for adult mice. J Virol 75:9121–9128 [View Article][PubMed]
    [Google Scholar]
  6. Ito N., Takayama-Ito M., Yamada K., Hosokawa J., Sugiyama M., Minamoto N. 2003; Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system. Microbiol Immunol 47:613–617[PubMed] [CrossRef]
    [Google Scholar]
  7. Ito K., Ishimaru T., Kimura F., Matsudomi N. 2007a; Importance of N-glycosylation positioning for secretion and folding of ovalbumin. Biochem Biophys Res Commun 361:725–731 [View Article][PubMed]
    [Google Scholar]
  8. Ito K., Seri A., Kimura F., Matsudomi N. 2007b; Site-specific glycosylation at Asn-292 in ovalbumin is essential to efficient secretion in yeast. J Biochem 141:193–199 [View Article][PubMed]
    [Google Scholar]
  9. Kasturi L., Eshleman J. R., Wunner W. H., Shakin-Eshleman S. H. 1995; The hydroxy amino acid in an Asn-X-Ser/Thr sequon can influence N-linked core glycosylation efficiency and the level of expression of a cell surface glycoprotein. J Biol Chem 270:14756–14761 [View Article][PubMed]
    [Google Scholar]
  10. Kayser V., Chennamsetty N., Voynov V., Forrer K., Helk B., Trout B. L. 2011; Glycosylation influences on the aggregation propensity of therapeutic monoclonal antibodies. Biotechnol J 6:38–44 [View Article][PubMed]
    [Google Scholar]
  11. Luo T. R., Minamoto N., Ito H., Goto H., Hiraga S., Ito N., Sugiyama M., Kinjo T. 1997; A virus-neutralizing epitope on the glycoprotein of rabies virus that contains Trp251 is a linear epitope. Virus Res 51:35–41 [View Article][PubMed]
    [Google Scholar]
  12. Shakin-Eshleman S. H., Remaley A. T., Eshleman J. R., Wunner W. H., Spitalnik S. L. 1992; N-linked glycosylation of rabies virus glycoprotein. Individual sequons differ in their glycosylation efficiencies and influence on cell surface expression. J Biol Chem 267:10690–10698[PubMed]
    [Google Scholar]
  13. Shakin-Eshleman S. H., Spitalnik S. L., Kasturi L. 1996; The amino acid at the X position of an Asn-X-Ser sequon is an important determinant of N-linked core-glycosylation efficiency. J Biol Chem 271:6363–6366 [View Article][PubMed]
    [Google Scholar]
  14. Shiota S., Mannen K., Matsumoto T., Yamada K., Yasui T., Takayama K., Kobayashi Y., Khawplod P., Gotoh K. other authors 2009; Development and evaluation of a rapid neutralizing antibody test for rabies. J Virol Methods 161:58–62 [View Article][PubMed]
    [Google Scholar]
  15. Somnuke P., Hauhart R. E., Atkinson J. P., Diamond M. S., Avirutnan P. 2011; N-linked glycosylation of dengue virus NS1 protein modulates secretion, cell-surface expression, hexamer stability, and interactions with human complement. Virology 413:253–264 [View Article][PubMed]
    [Google Scholar]
  16. Wunner W. H. 2007; Rabies virus. In Rabies, 2nd edn. pp. 23–68 Edited by Jackson A. C., Wunner W. H. San Diego, CA: Academic Press; [View Article]
    [Google Scholar]
  17. Wunner W. H., Dietzschold B., Smith C. L., Lafon M., Golub E. 1985; Antigenic variants of CVS rabies virus with altered glycosylation sites. Virology 140:1–12 [View Article][PubMed]
    [Google Scholar]
  18. Yamada K., Park C. H., Noguchi K., Kojima D., Kubo T., Komiya N., Matsumoto T., Mitui M. T., Ahmed K. other authors 2012; Serial passage of a street rabies virus in mouse neuroblastoma cells resulted in attenuation: potential role of the additional N-glycosylation of a viral glycoprotein in the reduced pathogenicity of street rabies virus. Virus Res 165:34–45 [View Article][PubMed]
    [Google Scholar]
  19. Zhou W., Tsai H. M. 2009; N-Glycans of ADAMTS13 modulate its secretion and von Willebrand factor cleaving activity. Blood 113:929–935 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.047852-0
Loading
/content/journal/jgv/10.1099/vir.0.047852-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
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