Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1989 Dec;63(12):5111–5118. doi: 10.1128/jvi.63.12.5111-5118.1989

Role of heterologous and homologous glycoproteins in phenotypic mixing between Sendai virus and vesicular stomatitis virus.

K Metsikkö 1, H Garoff 1
PMCID: PMC251173  PMID: 2555547

Abstract

Phenotypic mixing between Sendai virus and vesicular stomatitis virus (VSV) or the mutant VSV ts045 was studied. Conditions were optimized for double infection, as shown by immunofluorescence microscopy. Virions from double-infected cells were separated by sequential velocity and isopycnic gradient centrifugations. Two types of particles with mixed protein compositions were found. One type was VSV particles with Sendai virus spikes, i.e., phenotypically mixed particles. A second type was Sendai virus-VSV associations, which in plaque assays also behaved as phenotypically mixed particles. The ratio of VSV G protein to Sendai virus glycoproteins on the cell surface was varied, using the VSV mutant ts045 in double infections. Thus, different amounts of the VSV G protein were allowed to reach the cell surface at 32, 38, and 39 degrees C in Sendai virus-infected cells. However, a fixed number of Sendai virus spikes was always found in the ts045 virions. This represented 12 to 16% of the number of G proteins present in normal VSV. Furthermore, the yield of ts045 virions was radically reduced during double infection when the temperature was raised to block G-protein transport to the cell surface, suggesting that the Sendai virus glycoproteins were not able to compensate for G protein in budding. These results emphasize the role of the G protein in VSV assembly.

Full text

PDF
5113

Images in this article

5113Image
on p.5113
5113Image
on p.5113
5114Image
on p.5114
5115Image
on p.5115

Selected References

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

  1. Calafat J., Janssen H., Démant P., Hilgers J., Závada J. Specific selection of host cell glycoproteins during assembly of murine leukaemia virus and vesicular stomatitis virus: presence of Thy-1 glycoprotein and absence of H-2, Pgp-1 and T-200 glycoproteins on the envelopes of these virus particles. J Gen Virol. 1983 Jun;64(Pt 6):1241–1253. doi: 10.1099/0022-1317-64-6-1241. [DOI] [PubMed] [Google Scholar]
  2. Chan J. C., Hixson D. C., Bowen J. M. Detection of vesicular stomatitis virus (murine leukemia virus) pseudotypes by immunoelectron microscopy. Virology. 1978 Jul 1;88(1):171–176. doi: 10.1016/0042-6822(78)90120-4. [DOI] [PubMed] [Google Scholar]
  3. Chen S. S., Ariel N., Huang A. S. Membrane anchors of vesicular stomatitis virus: characterization and incorporation into virions. J Virol. 1988 Aug;62(8):2552–2556. doi: 10.1128/jvi.62.8.2552-2556.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Choppin P. W., Compans R. W. Phenotypic mixing of envelope proteins of the parainfluenza virus SV5 and vesicular stomatitis virus. J Virol. 1970 May;5(5):609–616. doi: 10.1128/jvi.5.5.609-616.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clapham P., Nagy K., Weiss R. A. Pseudotypes of human T-cell leukemia virus types 1 and 2: neutralization by patients' sera. Proc Natl Acad Sci U S A. 1984 May;81(9):2886–2889. doi: 10.1073/pnas.81.9.2886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Doxsey S. J., Sambrook J., Helenius A., White J. An efficient method for introducing macromolecules into living cells. J Cell Biol. 1985 Jul;101(1):19–27. doi: 10.1083/jcb.101.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gallione C. J., Rose J. K. A single amino acid substitution in a hydrophobic domain causes temperature-sensitive cell-surface transport of a mutant viral glycoprotein. J Virol. 1985 May;54(2):374–382. doi: 10.1128/jvi.54.2.374-382.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Griffiths G., Pfeiffer S., Simons K., Matlin K. Exit of newly synthesized membrane proteins from the trans cisterna of the Golgi complex to the plasma membrane. J Cell Biol. 1985 Sep;101(3):949–964. doi: 10.1083/jcb.101.3.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Homma M., Ouchi M. Trypsin action on the growth of Sendai virus in tissue culture cells. 3. Structural difference of Sendai viruses grown in eggs and tissue culture cells. J Virol. 1973 Dec;12(6):1457–1465. doi: 10.1128/jvi.12.6.1457-1465.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Knipe D. M., Baltimore D., Lodish H. F. Maturation of viral proteins in cells infected with temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1149–1158. doi: 10.1128/jvi.21.3.1149-1158.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lodish H. F., Porter M. Specific incorporation of host cell surface proteins into budding vesicular stomatitis virus particles. Cell. 1980 Jan;19(1):161–169. doi: 10.1016/0092-8674(80)90397-9. [DOI] [PubMed] [Google Scholar]
  13. Louvard D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4132–4136. doi: 10.1073/pnas.77.7.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Matlin K., Bainton D. F., Pesonen M., Louvard D., Genty N., Simons K. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence. J Cell Biol. 1983 Sep;97(3):627–637. doi: 10.1083/jcb.97.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McCloskey M., Poo M. M. Protein diffusion in cell membranes: some biological implications. Int Rev Cytol. 1984;87:19–81. doi: 10.1016/s0074-7696(08)62439-0. [DOI] [PubMed] [Google Scholar]
  16. McSharry J. J., Compans R. W., Choppin P. W. Proteins of vesicular stomatitis virus and of phenotypically mixed vesicular stomatitis virus-simian virus 5 virions. J Virol. 1971 Nov;8(5):722–729. doi: 10.1128/jvi.8.5.722-729.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Metsikkö K., Simons K. The budding mechanism of spikeless vesicular stomatitis virus particles. EMBO J. 1986 Aug;5(8):1913–1920. doi: 10.1002/j.1460-2075.1986.tb04444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nielsen B. L., Brown L. R. The basis for colored silver-protein complex formation in stained polyacrylamide gels. Anal Biochem. 1984 Sep;141(2):311–315. doi: 10.1016/0003-2697(84)90047-2. [DOI] [PubMed] [Google Scholar]
  19. Ogura H., Bauer H. Biological and electron microscopic studies on the phenotypic mixing of the thermolabile mutant of vesicular stomatitis virus, tl-17, with avian RNA tumor viruses. Arch Virol. 1976;52(3):233–242. doi: 10.1007/BF01348020. [DOI] [PubMed] [Google Scholar]
  20. Orvell C., Grandien M. The effects of monoclonal antibodies on biologic activities of structural proteins of Sendai virus. J Immunol. 1982 Dec;129(6):2779–2787. [PubMed] [Google Scholar]
  21. Orvell C., Rydbeck R., Löve A. Immunological relationships between mumps virus and parainfluenza viruses studied with monoclonal antibodies. J Gen Virol. 1986 Sep;67(Pt 9):1929–1939. doi: 10.1099/0022-1317-67-9-1929. [DOI] [PubMed] [Google Scholar]
  22. Pesonen M., Simons K. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. II. Immunological quantitation. J Cell Biol. 1983 Sep;97(3):638–643. doi: 10.1083/jcb.97.3.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rose J. K., Gallione C. J. Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus G and M proteins determined from cDNA clones containing the complete coding regions. J Virol. 1981 Aug;39(2):519–528. doi: 10.1128/jvi.39.2.519-528.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. RusS G., Poláková K., Závada J. Assembly of xenotropic murine leukaemia virus-related antigens from the surface of mouse L cells by vesicular stomatitis virus. Acta Virol. 1983 Mar;27(2):105–109. [PubMed] [Google Scholar]
  25. Scheid A., Choppin P. W. Two disulfide-linked polypeptide chains constitute the active F protein of paramyxoviruses. Virology. 1977 Jul 1;80(1):54–66. doi: 10.1016/0042-6822(77)90380-4. [DOI] [PubMed] [Google Scholar]
  26. Schnitzer T. J., Dickson C., Weiss R. A. Morphological and biochemical characterization of viral particles produced by the tsO45 mutant of vesicular stomatitis virus at restrictive temperature. J Virol. 1979 Jan;29(1):185–195. doi: 10.1128/jvi.29.1.185-195.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shioda T., Iwasaki K., Shibuta H. Determination of the complete nucleotide sequence of the Sendai virus genome RNA and the predicted amino acid sequences of the F, HN and L proteins. Nucleic Acids Res. 1986 Feb 25;14(4):1545–1563. doi: 10.1093/nar/14.4.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Thomas D., Newcomb W. W., Brown J. C., Wall J. S., Hainfeld J. F., Trus B. L., Steven A. C. Mass and molecular composition of vesicular stomatitis virus: a scanning transmission electron microscopy analysis. J Virol. 1985 May;54(2):598–607. doi: 10.1128/jvi.54.2.598-607.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vaux D. J., Helenius A., Mellman I. Spike--nucleocapsid interaction in Semliki Forest virus reconstructed using network antibodies. Nature. 1988 Nov 3;336(6194):36–42. doi: 10.1038/336036a0. [DOI] [PubMed] [Google Scholar]
  30. Weiss R. A., Bennett P. L. Assembly of membrane glycoproteins studied by phenotypic mixing between mutants of vesicular stomatitis virus and retroviruses. Virology. 1980 Jan 30;100(2):252–274. doi: 10.1016/0042-6822(80)90518-8. [DOI] [PubMed] [Google Scholar]
  31. Witte O. N., Baltimore D. Mechanism of formation of pseudotypes between vesicular stomatitis virus and murine leukemia virus. Cell. 1977 Jul;11(3):505–511. doi: 10.1016/0092-8674(77)90068-x. [DOI] [PubMed] [Google Scholar]
  32. Zilberstein A., Snider M. D., Porter M., Lodish H. F. Mutants of vesicular stomatitis virus blocked at different stages in maturation of the viral glycoprotein. Cell. 1980 Sep;21(2):417–427. doi: 10.1016/0092-8674(80)90478-x. [DOI] [PubMed] [Google Scholar]
  33. Závada J., Rosenbergová M. Phenotypic mixing of vesicular stomatitis virus with fowl plague virus. Acta Virol. 1972 Mar;16(2):103–114. [PubMed] [Google Scholar]
  34. Závada J. The pseudotypic paradox. J Gen Virol. 1982 Nov;63(Pt 1):15–24. doi: 10.1099/0022-1317-63-1-15. [DOI] [PubMed] [Google Scholar]
  35. van Meer G., Simons K. An efficient method for introducing defined lipids into the plasma membrane of mammalian cells. J Cell Biol. 1983 Nov;97(5 Pt 1):1365–1374. doi: 10.1083/jcb.97.5.1365. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES