Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jul;71(7):5487–5494. doi: 10.1128/jvi.71.7.5487-5494.1997

Formation of intracellular particles by hepatitis B virus large surface protein.

Z Xu 1, V Bruss 1, T S Yen 1
PMCID: PMC191790  PMID: 9188622

Abstract

Hepatitis B virus small surface protein is synthesized as a transmembrane protein of the rough endoplasmic reticulum (RER) and then buds into the lumen in the form of subviral particles that are secreted. The closely related large surface protein is also targeted to the RER but is retained in a pre-Golgi compartment and cannot be secreted. It has been assumed that the large surface protein remains as a transmembrane RER protein and hence cannot form particles, possibly because of binding to a host factor on the cytosolic face of the RER membranes. We have reexamined this question and found the following results. (i) The retained large surface protein is associated not with RER but, rather, with a more distal compartment. (ii) Electron microscopy reveals intravesicular 20-nm particles, similar to those formed by the small surface protein. (iii) The large surface protein colocalizes with and binds to calnexin, an ER chaperone protein. Therefore, our results indicate that the large surface protein is capable of budding and forming particles, and hence its intracellular retention cannot be attributed to a cytosolic factor. We interpret the data as evidence that the large surface protein is retained by virtue of interacting with calnexin, a component of what is considered the quality control mechanism of the ER.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bruss V., Lu X., Thomssen R., Gerlich W. H. Post-translational alterations in transmembrane topology of the hepatitis B virus large envelope protein. EMBO J. 1994 May 15;13(10):2273–2279. doi: 10.1002/j.1460-2075.1994.tb06509.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruss V., Thomssen R. Mapping a region of the large envelope protein required for hepatitis B virion maturation. J Virol. 1994 Mar;68(3):1643–1650. doi: 10.1128/jvi.68.3.1643-1650.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bruss V., Vieluf K. Functions of the internal pre-S domain of the large surface protein in hepatitis B virus particle morphogenesis. J Virol. 1995 Nov;69(11):6652–6657. doi: 10.1128/jvi.69.11.6652-6657.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chavrier P., Parton R. G., Hauri H. P., Simons K., Zerial M. Localization of low molecular weight GTP binding proteins to exocytic and endocytic compartments. Cell. 1990 Jul 27;62(2):317–329. doi: 10.1016/0092-8674(90)90369-p. [DOI] [PubMed] [Google Scholar]
  5. Cheng K. C., Smith G. L., Moss B. Hepatitis B virus large surface protein is not secreted but is immunogenic when selectively expressed by recombinant vaccinia virus. J Virol. 1986 Nov;60(2):337–344. doi: 10.1128/jvi.60.2.337-344.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chisari F. V., Filippi P., Buras J., McLachlan A., Popper H., Pinkert C. A., Palmiter R. D., Brinster R. L. Structural and pathological effects of synthesis of hepatitis B virus large envelope polypeptide in transgenic mice. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6909–6913. doi: 10.1073/pnas.84.19.6909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chisari F. V., Filippi P., McLachlan A., Milich D. R., Riggs M., Lee S., Palmiter R. D., Pinkert C. A., Brinster R. L. Expression of hepatitis B virus large envelope polypeptide inhibits hepatitis B surface antigen secretion in transgenic mice. J Virol. 1986 Dec;60(3):880–887. doi: 10.1128/jvi.60.3.880-887.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eble B. E., Lingappa V. R., Ganem D. Hepatitis B surface antigen: an unusual secreted protein initially synthesized as a transmembrane polypeptide. Mol Cell Biol. 1986 May;6(5):1454–1463. doi: 10.1128/mcb.6.5.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gallina A., Gazina E., Milanesi G. A C-terminal PreS1 sequence is sufficient to retain hepatitis B virus L protein in 293 cells. Virology. 1995 Oct 20;213(1):57–69. doi: 10.1006/viro.1995.1546. [DOI] [PubMed] [Google Scholar]
  10. Ganem D. Assembly of hepadnaviral virions and subviral particles. Curr Top Microbiol Immunol. 1991;168:61–83. doi: 10.1007/978-3-642-76015-0_4. [DOI] [PubMed] [Google Scholar]
  11. Ganem D., Varmus H. E. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. doi: 10.1146/annurev.bi.56.070187.003251. [DOI] [PubMed] [Google Scholar]
  12. Gerber M. A., Hadziyannis S., Vissoulis C., Schaffner F., Paronetto F., Popper H. Electron microscopy and immunoelectronmicroscopy of cytoplasmic hepatitis B antigen in hepatocytes. Am J Pathol. 1974 Jun;75(3):489–502. [PMC free article] [PubMed] [Google Scholar]
  13. Gething M. J., Sambrook J. Protein folding in the cell. Nature. 1992 Jan 2;355(6355):33–45. doi: 10.1038/355033a0. [DOI] [PubMed] [Google Scholar]
  14. Hammond C., Helenius A. Folding of VSV G protein: sequential interaction with BiP and calnexin. Science. 1994 Oct 21;266(5184):456–458. doi: 10.1126/science.7939687. [DOI] [PubMed] [Google Scholar]
  15. Huang Z. M., Yen T. S. Dysregulated surface gene expression from disrupted hepatitis B virus genomes. J Virol. 1993 Dec;67(12):7032–7040. doi: 10.1128/jvi.67.12.7032-7040.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huovila A. P., Eder A. M., Fuller S. D. Hepatitis B surface antigen assembles in a post-ER, pre-Golgi compartment. J Cell Biol. 1992 Sep;118(6):1305–1320. doi: 10.1083/jcb.118.6.1305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Le A., Steiner J. L., Ferrell G. A., Shaker J. C., Sifers R. N. Association between calnexin and a secretion-incompetent variant of human alpha 1-antitrypsin. J Biol Chem. 1994 Mar 11;269(10):7514–7519. [PubMed] [Google Scholar]
  18. Molnar-Kimber K. L., Jarocki-Witek V., Dheer S. K., Vernon S. K., Conley A. J., Davis A. R., Hung P. P. Distinctive properties of the hepatitis B virus envelope proteins. J Virol. 1988 Feb;62(2):407–416. doi: 10.1128/jvi.62.2.407-416.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ostapchuk P., Hearing P., Ganem D. A dramatic shift in the transmembrane topology of a viral envelope glycoprotein accompanies hepatitis B viral morphogenesis. EMBO J. 1994 Mar 1;13(5):1048–1057. doi: 10.1002/j.1460-2075.1994.tb06353.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ou J. H., Rutter W. J. Regulation of secretion of the hepatitis B virus major surface antigen by the preS-1 protein. J Virol. 1987 Mar;61(3):782–786. doi: 10.1128/jvi.61.3.782-786.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ou W. J., Cameron P. H., Thomas D. Y., Bergeron J. J. Association of folding intermediates of glycoproteins with calnexin during protein maturation. Nature. 1993 Aug 26;364(6440):771–776. doi: 10.1038/364771a0. [DOI] [PubMed] [Google Scholar]
  22. Persing D. H., Varmus H. E., Ganem D. Inhibition of secretion of hepatitis B surface antigen by a related presurface polypeptide. Science. 1986 Dec 12;234(4782):1388–1391. doi: 10.1126/science.3787251. [DOI] [PubMed] [Google Scholar]
  23. Prange R., Nagel R., Streeck R. E. Deletions in the hepatitis B virus small envelope protein: effect on assembly and secretion of surface antigen particles. J Virol. 1992 Oct;66(10):5832–5841. doi: 10.1128/jvi.66.10.5832-5841.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Prange R., Streeck R. E. Novel transmembrane topology of the hepatitis B virus envelope proteins. EMBO J. 1995 Jan 16;14(2):247–256. doi: 10.1002/j.1460-2075.1995.tb06998.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rajagopalan S., Xu Y., Brenner M. B. Retention of unassembled components of integral membrane proteins by calnexin. Science. 1994 Jan 21;263(5145):387–390. doi: 10.1126/science.8278814. [DOI] [PubMed] [Google Scholar]
  26. Schweizer A., Matter K., Ketcham C. M., Hauri H. P. The isolated ER-Golgi intermediate compartment exhibits properties that are different from ER and cis-Golgi. J Cell Biol. 1991 Apr;113(1):45–54. doi: 10.1083/jcb.113.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Simon K., Lingappa V. R., Ganem D. Secreted hepatitis B surface antigen polypeptides are derived from a transmembrane precursor. J Cell Biol. 1988 Dec;107(6 Pt 1):2163–2168. doi: 10.1083/jcb.107.6.2163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Traavik T., Kjeldsberg E., Siebke J. C. The effects of detergent-treatment on the morphology of Australia antigen positive particles. Acta Pathol Microbiol Scand B Microbiol Immunol. 1973 Feb;81(1):37–42. doi: 10.1111/j.1699-0463.1973.tb02184.x. [DOI] [PubMed] [Google Scholar]
  29. Wang C. J., Chen P. J., Wu J. C., Patel D., Chen D. S. Small-form hepatitis B surface antigen is sufficient to help in the assembly of hepatitis delta virus-like particles. J Virol. 1991 Dec;65(12):6630–6636. doi: 10.1128/jvi.65.12.6630-6636.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wieland F. T., Gleason M. L., Serafini T. A., Rothman J. E. The rate of bulk flow from the endoplasmic reticulum to the cell surface. Cell. 1987 Jul 17;50(2):289–300. doi: 10.1016/0092-8674(87)90224-8. [DOI] [PubMed] [Google Scholar]
  31. Xu Z., Yen T. S. Intracellular retention of surface protein by a hepatitis B virus mutant that releases virion particles. J Virol. 1996 Jan;70(1):133–140. doi: 10.1128/jvi.70.1.133-140.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES