Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Oct 15;93(21):11359–11365. doi: 10.1073/pnas.93.21.11359

Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles.

M J Schnell 1, L Buonocore 1, E Kretzschmar 1, E Johnson 1, J K Rose 1
PMCID: PMC38062  PMID: 8876140

Abstract

In a previous study we demonstrated that vesicular stomatitis virus (VSV) can be used as a vector to express a soluble protein in mammalian cells. Here we have generated VSV recombinants that express four different membrane proteins: the cellular CD4 protein, a CD4-G hybrid protein containing the ectodomain of CD4 and the transmembrane and cytoplasmic tail of the VSV glycoprotein (G), the measles virus hemagglutinin, or the measles virus fusion protein. The proteins were expressed at levels ranging from 23-62% that of VSV G protein and all were transported to the cell surface. In addition we found that all four proteins were incorporated into the membrane envelope of VSV along with the VSV G protein. The levels of incorporation of these proteins varied from 6-31% of that observed for VSV G. These results suggest that many different membrane proteins may be co-incorporated quite efficiently with VSV G protein into budding VSV virus particles and that specific signals are not required for this co-incorporation process. In fact, the CD4-G protein was incorporated with the same efficiency as wild type CD4. Electron microscopy of virions containing CD4 revealed that the CD4 molecules were dispersed throughout the virion envelope among the trimeric viral spike glycoproteins. The recombinant VSV-CD4 virus particles were about 18% longer than wild type virions, reflecting the additional length of the helical nucleocapsid containing the extra gene. Recombinant VSVs carrying foreign antigens on the surface of the virus particle may be useful for viral targeting, membrane protein purification, and for generation of immune responses.

Full text

PDF
11361

Images in this article

11360Fig. 1
on p.11360
11361Fig. 2
on p.11361
11362Fig. 3
on p.11362
11363Fig. 4
on p.11363
11363Fig. 6
on p.11363

Selected References

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

  1. Cattaneo R., Rose J. K. Cell fusion by the envelope glycoproteins of persistent measles viruses which caused lethal human brain disease. J Virol. 1993 Mar;67(3):1493–1502. doi: 10.1128/jvi.67.3.1493-1502.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chong L. D., Rose J. K. Interactions of normal and mutant vesicular stomatitis virus matrix proteins with the plasma membrane and nucleocapsids. J Virol. 1994 Jan;68(1):441–447. doi: 10.1128/jvi.68.1.441-447.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chong L. D., Rose J. K. Membrane association of functional vesicular stomatitis virus matrix protein in vivo. J Virol. 1993 Jan;67(1):407–414. doi: 10.1128/jvi.67.1.407-414.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fuerst T. R., Niles E. G., Studier F. W., Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. doi: 10.1073/pnas.83.21.8122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Guan J. L., Cao H., Rose J. K. Cell-surface expression of a membrane-anchored form of the human chorionic gonadotropin alpha subunit. J Biol Chem. 1988 Apr 15;263(11):5306–5313. [PubMed] [Google Scholar]
  6. Justice P. A., Sun W., Li Y., Ye Z., Grigera P. R., Wagner R. R. Membrane vesiculation function and exocytosis of wild-type and mutant matrix proteins of vesicular stomatitis virus. J Virol. 1995 May;69(5):3156–3160. doi: 10.1128/jvi.69.5.3156-3160.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Lefrancois L., Lyles D. S. The interaction of antibody with the major surface glycoprotein of vesicular stomatitis virus. II. Monoclonal antibodies of nonneutralizing and cross-reactive epitopes of Indiana and New Jersey serotypes. Virology. 1982 Aug;121(1):168–174. doi: 10.1016/0042-6822(82)90126-x. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell. 1986 Nov 7;47(3):333–348. doi: 10.1016/0092-8674(86)90590-8. [DOI] [PubMed] [Google Scholar]
  11. Mebatsion T., Konig M., Conzelmann K. K. Budding of rabies virus particles in the absence of the spike glycoprotein. Cell. 1996 Mar 22;84(6):941–951. doi: 10.1016/s0092-8674(00)81072-7. [DOI] [PubMed] [Google Scholar]
  12. Owens R. J., Rose J. K. Cytoplasmic domain requirement for incorporation of a foreign envelope protein into vesicular stomatitis virus. J Virol. 1993 Jan;67(1):360–365. doi: 10.1128/jvi.67.1.360-365.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Reinherz E. L., Kung P. C., Goldstein G., Schlossman S. F. Separation of functional subsets of human T cells by a monoclonal antibody. Proc Natl Acad Sci U S A. 1979 Aug;76(8):4061–4065. doi: 10.1073/pnas.76.8.4061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rose J. K., Buonocore L., Whitt M. A. A new cationic liposome reagent mediating nearly quantitative transfection of animal cells. Biotechniques. 1991 Apr;10(4):520–525. [PubMed] [Google Scholar]
  15. Rose R. M. The role of colony-stimulating factors in infectious disease: current status, future challenges. Semin Oncol. 1992 Aug;19(4):415–421. [PubMed] [Google Scholar]
  16. Schnell M. J., Buonocore L., Whitt M. A., Rose J. K. The minimal conserved transcription stop-start signal promotes stable expression of a foreign gene in vesicular stomatitis virus. J Virol. 1996 Apr;70(4):2318–2323. doi: 10.1128/jvi.70.4.2318-2323.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schnitzer T. J., Lodish H. F. Noninfectious vesicular stomatitis virus particles deficient in the viral nucleocapsid. J Virol. 1979 Feb;29(2):443–447. doi: 10.1128/jvi.29.2.443-447.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schubert M., Joshi B., Blondel D., Harmison G. G. Insertion of the human immunodeficiency virus CD4 receptor into the envelope of vesicular stomatitis virus particles. J Virol. 1992 Mar;66(3):1579–1589. doi: 10.1128/jvi.66.3.1579-1589.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shaw A. S., Amrein K. E., Hammond C., Stern D. F., Sefton B. M., Rose J. K. The lck tyrosine protein kinase interacts with the cytoplasmic tail of the CD4 glycoprotein through its unique amino-terminal domain. Cell. 1989 Nov 17;59(4):627–636. doi: 10.1016/0092-8674(89)90008-1. [DOI] [PubMed] [Google Scholar]
  20. Whitt M. A., Chong L., Rose J. K. Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant. J Virol. 1989 Sep;63(9):3569–3578. doi: 10.1128/jvi.63.9.3569-3578.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Young J. A., Bates P., Willert K., Varmus H. E. Efficient incorporation of human CD4 protein into avian leukosis virus particles. Science. 1990 Dec 7;250(4986):1421–1423. doi: 10.1126/science.2175047. [DOI] [PubMed] [Google Scholar]
  23. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES