Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1992 Dec;66(12):7089–7095. doi: 10.1128/jvi.66.12.7089-7095.1992

Role of cell surface spikes in alphavirus budding.

H Zhao 1, H Garoff 1
PMCID: PMC240383  PMID: 1331511

Abstract

Alphaviruses mature by budding at cell surfaces. According to a prevailing hypothesis, the viral membrane protein, which is a heterodimeric protein unit, is transported to the plasma membrane (PM), where it awaits binding to the viral nucleocapsid (NC). This hypothesis predicts that the viral membrane protein heterodimers accumulate at the cell surface when expressed in the absence of NCs. We have tested this prediction by analyzing the spike protein expression phenotype of a Semliki Forest virus (SFV) variant which contains a capsid gene deletion. We found that viral membrane protein heterodimers were formed and transported to the cell surface normally. However, instead of accumulating at the PM as expected, the membrane proteins were rapidly degraded. In the case of the E1 subunit, degradation resulted in the release of a soluble E1 fragment into the medium. The fact that this pathway of protein degradation is mostly inhibited during wild-type virus infection suggests that viral membrane proteins are very efficiently captured by NCs into budding complexes and that normally no sizeable pool of free membrane protein complexes exists at the PM.

Full text

PDF
7089

Images in this article

7091Image
on p.7091
7092Image
on p.7092
7092Image
on p.7092
7093Image
on p.7093
7093Image
on p.7093
7093Image
on p.7093

Selected References

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

  1. Boere W. A., Harmsen T., Vinjé J., Benaissa-Trouw B. J., Kraaijeveld C. A., Snippe H. Identification of distinct antigenic determinants on Semliki Forest virus by using monoclonal antibodies with different antiviral activities. J Virol. 1984 Nov;52(2):575–582. doi: 10.1128/jvi.52.2.575-582.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruss V., Ganem D. The role of envelope proteins in hepatitis B virus assembly. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):1059–1063. doi: 10.1073/pnas.88.3.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Delchambre M., Gheysen D., Thines D., Thiriart C., Jacobs E., Verdin E., Horth M., Burny A., Bex F. The GAG precursor of simian immunodeficiency virus assembles into virus-like particles. EMBO J. 1989 Sep;8(9):2653–2660. doi: 10.1002/j.1460-2075.1989.tb08405.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Garoff H., Huylebroeck D., Robinson A., Tillman U., Liljeström P. The signal sequence of the p62 protein of Semliki Forest virus is involved in initiation but not in completing chain translocation. J Cell Biol. 1990 Sep;111(3):867–876. doi: 10.1083/jcb.111.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garoff H., Kondor-Koch C., Riedel H. Structure and assembly of alphaviruses. Curr Top Microbiol Immunol. 1982;99:1–50. doi: 10.1007/978-3-642-68528-6_1. [DOI] [PubMed] [Google Scholar]
  6. Green J., Griffiths G., Louvard D., Quinn P., Warren G. Passage of viral membrane proteins through the Golgi complex. J Mol Biol. 1981 Nov 15;152(4):663–698. doi: 10.1016/0022-2836(81)90122-4. [DOI] [PubMed] [Google Scholar]
  7. Johnson D. C., Schlesinger M. J., Elson E. L. Fluorescence photobleaching recovery measurements reveal differences in envelopment of Sindbis and vesicular stomatitis viruses. Cell. 1981 Feb;23(2):423–431. doi: 10.1016/0092-8674(81)90137-9. [DOI] [PubMed] [Google Scholar]
  8. Karacostas V., Nagashima K., Gonda M. A., Moss B. Human immunodeficiency virus-like particles produced by a vaccinia virus expression vector. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8964–8967. doi: 10.1073/pnas.86.22.8964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Liljeström P., Garoff H. A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology (N Y) 1991 Dec;9(12):1356–1361. doi: 10.1038/nbt1291-1356. [DOI] [PubMed] [Google Scholar]
  10. Lobigs M., Zhao H. X., Garoff H. Function of Semliki Forest virus E3 peptide in virus assembly: replacement of E3 with an artificial signal peptide abolishes spike heterodimerization and surface expression of E1. J Virol. 1990 Sep;64(9):4346–4355. doi: 10.1128/jvi.64.9.4346-4355.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mayne J. T., Rice C. M., Strauss E. G., Hunkapiller M. W., Strauss J. H. Biochemical studies of the maturation of the small Sindbis virus glycoprotein E3. Virology. 1984 Apr 30;134(2):338–357. doi: 10.1016/0042-6822(84)90302-7. [DOI] [PubMed] [Google Scholar]
  12. Pettersson R. F. Protein localization and virus assembly at intracellular membranes. Curr Top Microbiol Immunol. 1991;170:67–106. doi: 10.1007/978-3-642-76389-2_3. [DOI] [PubMed] [Google Scholar]
  13. Rhee S. S., Hui H. X., Hunter E. Preassembled capsids of type D retroviruses contain a signal sufficient for targeting specifically to the plasma membrane. J Virol. 1990 Aug;64(8):3844–3852. doi: 10.1128/jvi.64.8.3844-3852.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Scheele C. M., Pfefferkorn E. R. Kinetics of incorporation of structural proteins into Sindbis virions. J Virol. 1969 Apr;3(4):369–375. doi: 10.1128/jvi.3.4.369-375.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Simons K., Garoff H. The budding mechanisms of enveloped animal viruses. J Gen Virol. 1980 Sep;50(1):1–21. doi: 10.1099/0022-1317-50-1-1. [DOI] [PubMed] [Google Scholar]
  16. Suomalainen M., Liljeström P., Garoff H. Spike protein-nucleocapsid interactions drive the budding of alphaviruses. J Virol. 1992 Aug;66(8):4737–4747. doi: 10.1128/jvi.66.8.4737-4747.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wahlberg J. M., Boere W. A., Garoff H. The heterodimeric association between the membrane proteins of Semliki Forest virus changes its sensitivity to low pH during virus maturation. J Virol. 1989 Dec;63(12):4991–4997. doi: 10.1128/jvi.63.12.4991-4997.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ziemiecki A., Garoff H., Simons K. Formation of the Semliki Forest virus membrane glycoprotein complexes in the infected cell. J Gen Virol. 1980 Sep;50(1):111–123. doi: 10.1099/0022-1317-50-1-111. [DOI] [PubMed] [Google Scholar]
  19. de Curtis I., Simons K. Dissection of Semliki Forest virus glycoprotein delivery from the trans-Golgi network to the cell surface in permeabilized BHK cells. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8052–8056. doi: 10.1073/pnas.85.21.8052. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES