1887

Abstract

The middle surface antigen (M-sAg) of hepadnaviruses is one of three envelope proteins that share a common C-terminal S domain. M-sAg contains the preS2 domain in addition to the S region. The preS2 region of woodchuck hepatitis virus (WHV) contains a potential glycosylation site Asn-Gln-Thr at amino acid (aa) positions 3–5. In this study, we mutated this site by site-directed mutagenesis and confirmed that glycosylation occurs here. In translation assays, the mutation Thr to Asn at aa 5 significantly impaired glycosylation of M-sAg. The mutated M-sAg formed abnormal clustered structures in transfected cells as determined by immunofluorescent staining. Confocal microscopic analysis showed that an enrichment of this glycosylation-deficient protein in the Golgi apparatus occurred, which is not typical for the wild-type protein. These results are consistent with earlier findings that incorrect glycosylation of viral proteins may interfere with virus assembly.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.19672-0
2004-04-01
2020-10-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/4/vir850787.html?itemId=/content/journal/jgv/10.1099/vir.0.19672-0&mimeType=html&fmt=ahah

References

  1. Block T. M., Lu X., Mehta A. S. 7 other authors 1998; Treatment of chronic hepadnavirus infection in a woodchuck animal model with an inhibitor of protein folding and trafficking. Nat Med 4:610–614
    [Google Scholar]
  2. Breiner K. M., Schaller H., Knolle P. A. 2001; Endothelial cell-mediated uptake of a hepatitis B virus: a new concept of liver targeting of hepatotropic microorganisms. Hepatology 34:803–808
    [Google Scholar]
  3. Chouteau P., Le Seyec J., Cannie I., Nassal M., Guguen-Guillouzo C., Gripon P. 2001; A short N-proximal region in the large envelope protein harbors a determinant that contributes to the species specificity of human hepatitis B virus. J Virol 75:1565–1572
    [Google Scholar]
  4. Farquhar M. G., Palade G. E. 1998; The Golgi apparatus: 100 years of progress and controversy. Trends Cell Biol 8:2–10
    [Google Scholar]
  5. Galibert F., Chen T. N., Mandart E. 1982; Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence. J Virol 41:51–65
    [Google Scholar]
  6. Le Seyec J., Chouteau P., Cannie I., Guguen-Guillouzo C., Gripon P. 1999; Infection process of the hepatitis B virus depends on the presence of a defined sequence in the pre-S1 domain. J Virol 73:2052–2057
    [Google Scholar]
  7. Lu X., Mehta A., Dadmarz M., Dwek R., Blumberg B. S., Block T. M. 1997; Aberrant trafficking of hepatitis B virus glycoproteins in cells in which N -glycan processing is inhibited. Proc Natl Acad Sci U S A 94:2380–2385
    [Google Scholar]
  8. Mehta A., Lu X., Block T. M., Blumberg B. S., Dwek R. A. 1997; Hepatitis B virus (HBV) envelope glycoproteins vary drastically in their sensitivity to glycan processing: evidence that alteration of a single N -linked glycosylation site can regulate HBV secretion. Proc Natl Acad Sci U S A 94:1822–1827
    [Google Scholar]
  9. Munro S. 1998; Localization of proteins to the Golgi apparatus. Trends Cell Biol 8:11–15
    [Google Scholar]
  10. Pollicino T., Zanetti A. R., Cacciola I. 8 other authors 1997; PreS2 defective hepatitis B virus infection in patients with fulminant hepatitis. Hepatology 26:495–499
    [Google Scholar]
  11. Rickwood D. 1992 Preparative Centrifugation – a Practical Approach Edited by Rickwood D. Oxford: Oxford University Press;
    [Google Scholar]
  12. Schmitt S., Glebe D., Alving K. 7 other authors 1999; Analysis of the pre-S2 N - and O -linked glycans of the M surface protein from human hepatitis B virus. J Biol Chem 274:11945–11957
    [Google Scholar]
  13. Stoll-Becker S., Repp R., Glebe D., Schaefer S., Kreuder J., Kann M., Lampert F., Gerlich W. H. 1997; Transcription of hepatitis B virus in peripheral blood mononuclear cells from persistently infected patients. J Virol 71:5399–5407
    [Google Scholar]
  14. Tolle T. K., Glebe D., Linder M., Linder D., Schmitt S., Geyer R., Gerlich W. H. 1998; Structure and glycosylation patterns of surface proteins from woodchuck hepatitis virus. J Virol 72:9978–9985
    [Google Scholar]
  15. Urban S., Breiner K. M., Fehler F., Klingmuller U., Schaller H. 1998; Avian hepatitis B virus infection is initiated by the interaction of a distinct pre-S subdomain with the cellular receptor gp180. J Virol 72:8089–8097
    [Google Scholar]
  16. Zheng X., Schirmbeck R., Hilken G., Waters J. A., Yang D., Reimann J., Roggendorf M., Lu M. 2002; Characterization of complex B-cell epitopes on woodchuck hepatitis virus surface antigens by using plasmids encoding chimeric proteins and DNA immunization. Virology 294:342–353
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.19672-0
Loading
/content/journal/jgv/10.1099/vir.0.19672-0
Loading

Data & Media loading...

Most cited this month 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