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. 1994 Mar 1;13(5):1058–1064. doi: 10.1002/j.1460-2075.1994.tb06354.x

Membrane protein lateral interactions control Semliki Forest virus budding.

M Ekström 1, P Liljeström 1, H Garoff 1
PMCID: PMC394913  PMID: 8131740

Abstract

Semliki Forest virus, SFV, directs the synthesis of two membrane proteins, p62 and E1, which form a p62E1 heterodimer in the endoplasmic reticulum. After being transported to the plasma membrane (PM), they are incorporated into the virus membrane during the process of virus budding. Electronmicroscopic analyses of the envelope in matured virus show that the heterodimers are clustered into trimeric structures (spikes) which further form a regular surface lattice with T = 4. In this work we have used a genetic approach to study the importance of the trimerization event for virus budding. We have coexpressed a budding competent form of the virus heterodimer with another one which cannot be used for particle formation because of a defect in nucleocapsid (NC) binding. We show that the NC binding-deficient heterodimer is able to inhibit the budding of the competent one in a concentration-dependent manner and that the NC binding-competent heterodimers can rescue the incompetent ones into virus particles. This suggests that the heterodimers are complexed together, probably into the trimeric structures (spikes), at the PM to expose a multivalent binding site for the NC and thereby drive efficient virus budding.

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Selected References

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

  1. Anthony R. P., Brown D. T. Protein-protein interactions in an alphavirus membrane. J Virol. 1991 Mar;65(3):1187–1194. doi: 10.1128/jvi.65.3.1187-1194.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barth B. U., Suomalainen M., Liljeström P., Garoff H. Alphavirus assembly and entry: role of the cytoplasmic tail of the E1 spike subunit. J Virol. 1992 Dec;66(12):7560–7564. doi: 10.1128/jvi.66.12.7560-7564.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berglund P., Sjöberg M., Garoff H., Atkins G. J., Sheahan B. J., Liljeström P. Semliki Forest virus expression system: production of conditionally infectious recombinant particles. Biotechnology (N Y) 1993 Aug;11(8):916–920. doi: 10.1038/nbt0893-916. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Bron R., Wahlberg J. M., Garoff H., Wilschut J. Membrane fusion of Semliki Forest virus in a model system: correlation between fusion kinetics and structural changes in the envelope glycoprotein. EMBO J. 1993 Feb;12(2):693–701. doi: 10.1002/j.1460-2075.1993.tb05703.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Burge B. W., Pfefferkorn E. R. Complementation between temperature-sensitive mutants of Sindbis virus. Virology. 1966 Oct;30(2):214–223. doi: 10.1016/0042-6822(66)90097-3. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Doms R. W., Lamb R. A., Rose J. K., Helenius A. Folding and assembly of viral membrane proteins. Virology. 1993 Apr;193(2):545–562. doi: 10.1006/viro.1993.1164. [DOI] [PubMed] [Google Scholar]
  10. Edwards J., Mann E., Brown D. T. Conformational changes in Sindbis virus envelope proteins accompanying exposure to low pH. J Virol. 1983 Mar;45(3):1090–1097. doi: 10.1128/jvi.45.3.1090-1097.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Flynn D. C., Meyer W. J., Mackenzie J. M., Jr, Johnston R. E. A conformational change in Sindbis virus glycoproteins E1 and E2 is detected at the plasma membrane as a consequence of early virus-cell interaction. J Virol. 1990 Aug;64(8):3643–3653. doi: 10.1128/jvi.64.8.3643-3653.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fuller S. D. The T=4 envelope of Sindbis virus is organized by interactions with a complementary T=3 capsid. Cell. 1987 Mar 27;48(6):923–934. doi: 10.1016/0092-8674(87)90701-x. [DOI] [PubMed] [Google Scholar]
  13. Gaedigk-Nitschko K., Schlesinger M. J. Site-directed mutations in Sindbis virus E2 glycoprotein's cytoplasmic domain and the 6K protein lead to similar defects in virus assembly and budding. Virology. 1991 Jul;183(1):206–214. doi: 10.1016/0042-6822(91)90133-v. [DOI] [PubMed] [Google Scholar]
  14. Garoff H., Frischauf A. M., Simons K., Lehrach H., Delius H. Nucleotide sequence of cdna coding for Semliki Forest virus membrane glycoproteins. Nature. 1980 Nov 20;288(5788):236–241. doi: 10.1038/288236a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Gheysen D., Jacobs E., de Foresta F., Thiriart C., Francotte M., Thines D., De Wilde M. Assembly and release of HIV-1 precursor Pr55gag virus-like particles from recombinant baculovirus-infected insect cells. Cell. 1989 Oct 6;59(1):103–112. doi: 10.1016/0092-8674(89)90873-8. [DOI] [PubMed] [Google Scholar]
  17. Griffiths G., Fuller S. D., Back R., Hollinshead M., Pfeiffer S., Simons K. The dynamic nature of the Golgi complex. J Cell Biol. 1989 Feb;108(2):277–297. doi: 10.1083/jcb.108.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Kielian M., Helenius A. pH-induced alterations in the fusogenic spike protein of Semliki Forest virus. J Cell Biol. 1985 Dec;101(6):2284–2291. doi: 10.1083/jcb.101.6.2284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liljeström P., Lusa S., Huylebroeck D., Garoff H. In vitro mutagenesis of a full-length cDNA clone of Semliki Forest virus: the small 6,000-molecular-weight membrane protein modulates virus release. J Virol. 1991 Aug;65(8):4107–4113. doi: 10.1128/jvi.65.8.4107-4113.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lobigs M., Garoff H. Fusion function of the Semliki Forest virus spike is activated by proteolytic cleavage of the envelope glycoprotein precursor p62. J Virol. 1990 Mar;64(3):1233–1240. doi: 10.1128/jvi.64.3.1233-1240.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Omar A., Koblet H. The use of sulfite to study the mechanism of membrane fusion induced by E1 of Semliki Forest virus. Virology. 1989 Jan;168(1):177–179. doi: 10.1016/0042-6822(89)90418-2. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Salminen A., Wahlberg J. M., Lobigs M., Liljeström P., Garoff H. Membrane fusion process of Semliki Forest virus. II: Cleavage-dependent reorganization of the spike protein complex controls virus entry. J Cell Biol. 1992 Jan;116(2):349–357. doi: 10.1083/jcb.116.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Simons K., Helenius A., Garoff H. Solubilization of the membrane proteins from Semliki Forest virus with Triton X100. J Mol Biol. 1973 Oct 15;80(1):119–133. doi: 10.1016/0022-2836(73)90236-2. [DOI] [PubMed] [Google Scholar]
  29. Snijders A., Benaissa-Trouw B. J., Oosterlaken T. A., Puijk W. C., Posthumus W. P., Meloen R. H., Boere W. A., Oosting J. D., Kraaijeveld C. A., Snippe H. Identification of linear epitopes on Semliki Forest virus E2 membrane protein and their effectiveness as a synthetic peptide vaccine. J Gen Virol. 1991 Mar;72(Pt 3):557–565. doi: 10.1099/0022-1317-72-3-557. [DOI] [PubMed] [Google Scholar]
  30. Strauss E. G., Tsukeda H., Simizu B. Mutants of sindbis virus. IV. Heterotypic complementation and phenotypic mixing between temperature-sensitive mutants and wild-type Sindbis and Western equine encephalitis viruses. J Gen Virol. 1983 Jul;64(Pt 7):1581–1590. doi: 10.1099/0022-1317-64-7-1581. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Vogel R. H., Provencher S. W., von Bonsdorff C. H., Adrian M., Dubochet J. Envelope structure of Semliki Forest virus reconstructed from cryo-electron micrographs. Nature. 1986 Apr 10;320(6062):533–535. doi: 10.1038/320533a0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Wahlberg J. M., Bron R., Wilschut J., Garoff H. Membrane fusion of Semliki Forest virus involves homotrimers of the fusion protein. J Virol. 1992 Dec;66(12):7309–7318. doi: 10.1128/jvi.66.12.7309-7318.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wahlberg J. M., Garoff H. Membrane fusion process of Semliki Forest virus. I: Low pH-induced rearrangement in spike protein quaternary structure precedes virus penetration into cells. J Cell Biol. 1992 Jan;116(2):339–348. doi: 10.1083/jcb.116.2.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wengler G., Wengler G. Cell-associated West Nile flavivirus is covered with E+pre-M protein heterodimers which are destroyed and reorganized by proteolytic cleavage during virus release. J Virol. 1989 Jun;63(6):2521–2526. doi: 10.1128/jvi.63.6.2521-2526.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wills J. W., Craven R. C., Achacoso J. A. Creation and expression of myristylated forms of Rous sarcoma virus gag protein in mammalian cells. J Virol. 1989 Oct;63(10):4331–4343. doi: 10.1128/jvi.63.10.4331-4343.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zhao H., Garoff H. Role of cell surface spikes in alphavirus budding. J Virol. 1992 Dec;66(12):7089–7095. doi: 10.1128/jvi.66.12.7089-7095.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Ziemiecki A., Garofff H. Subunit composition of the membrane glycoprotein complex of Semliki Forest virus. J Mol Biol. 1978 Jul 5;122(3):259–269. doi: 10.1016/0022-2836(78)90189-4. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. von Bonsdorff C. H., Harrison S. C. Hexagonal glycoprotein arrays from Sindbis virus membranes. J Virol. 1978 Nov;28(2):578–583. doi: 10.1128/jvi.28.2.578-583.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

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