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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 Aug 6;93(16):8268–8273. doi: 10.1073/pnas.93.16.8268

Migration of vesicular stomatitis virus glycoprotein to the nucleus of infected cells.

A T Da Poian 1, A M Gomes 1, R J Oliveira 1, J L Silva 1
PMCID: PMC38659  PMID: 8710859

Abstract

A new means of direct visualization of the early events of viral infection by selective fluorescence labeling of viral proteins coupled with digital imaging microscopy is reported. The early phases of viral infection have great importance for understanding viral replication and pathogenesis. Vesicular stomatitis virus, the best-studied rhabdovirus, is composed of an RNA genome of negative sense, five viral proteins, and membrane lipids derived from the host cell. The glycoprotein of vesicular stomatitis virus was labeled with fluorescein isothiocyanate, and the labeled virus was incubated with baby hamster kidney cells. After initiation of infection, the fluorescence of the labeled glycoprotein was first seen inside the cells in endocytic vesicles. The fluorescence progressively migrated to the nucleus of infected cells. After 1 h of infection, the virus glycoprotein was concentrated in the nucleus and could be recovered intact in a preparation of purified nuclei. These results suggest that uncoating of the viral RNA occurs close to the nuclear membrane, which would precede transcription of the leader RNA that enters the nucleus to shut off cellular RNA synthesis and DNA replication.

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Selected References

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

  1. Baltimore D., Huang A. S., Stampfer M. Ribonucleic acid synthesis of vesicular stomatitis virus, II. An RNA polymerase in the virion. Proc Natl Acad Sci U S A. 1970 Jun;66(2):572–576. doi: 10.1073/pnas.66.2.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bishop D. H., Repik P., Obijeski J. F., Moore N. F., Wagner R. R. Restitution of infectivity to spikeless vesicular stomatitis virus by solubilized viral components. J Virol. 1975 Jul;16(1):75–84. doi: 10.1128/jvi.16.1.75-84.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown F., Cartwright B. The antigens of vesicular stomatitis virus. II. The presence of two low molecular weight immunogens in virus suspensions. J Immunol. 1966 Nov;97(5):612–620. [PubMed] [Google Scholar]
  4. De Robertis E. M., Lienhard S., Parisot R. F. Intracellular transport of microinjected 5S and small nuclear RNAs. Nature. 1982 Feb 18;295(5850):572–577. doi: 10.1038/295572a0. [DOI] [PubMed] [Google Scholar]
  5. Dietzschold B., Tollis M., Lafon M., Wunner W. H., Koprowski H. Mechanisms of rabies virus neutralization by glycoprotein-specific monoclonal antibodies. Virology. 1987 Nov;161(1):29–36. doi: 10.1016/0042-6822(87)90167-x. [DOI] [PubMed] [Google Scholar]
  6. Florkiewicz R. Z., Smith A., Bergmann J. E., Rose J. K. Isolation of stable mouse cell lines that express cell surface and secreted forms of the vesicular stomatitis virus glycoprotein. J Cell Biol. 1983 Nov;97(5 Pt 1):1381–1388. doi: 10.1083/jcb.97.5.1381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Follett E. A., Pringle C. R., Wunner W. H., Skehel J. J. Virus replication in enucleate cells: vesicular stomatitis virus and influenza virus. J Virol. 1974 Feb;13(2):394–399. doi: 10.1128/jvi.13.2.394-399.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Forbes D. J. Structure and function of the nuclear pore complex. Annu Rev Cell Biol. 1992;8:495–527. doi: 10.1146/annurev.cb.08.110192.002431. [DOI] [PubMed] [Google Scholar]
  9. Gollins S. W., Porterfield J. S. A new mechanism for the neutralization of enveloped viruses by antiviral antibody. Nature. 1986 May 15;321(6067):244–246. doi: 10.1038/321244a0. [DOI] [PubMed] [Google Scholar]
  10. Grinnell B. W., Wagner R. R. Nucleotide sequence and secondary structure of VSV leader RNA and homologous DNA involved in inhibition of DNA-dependent transcription. Cell. 1984 Feb;36(2):533–543. doi: 10.1016/0092-8674(84)90246-0. [DOI] [PubMed] [Google Scholar]
  11. Helenius A. Unpacking the incoming influenza virus. Cell. 1992 May 15;69(4):577–578. doi: 10.1016/0092-8674(92)90219-3. [DOI] [PubMed] [Google Scholar]
  12. Kurilla M. G., Piwnica-Worms H., Keene J. D. Rapid and transient localization of the leader RNA of vesicular stomatitis virus in the nuclei of infected cells. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5240–5244. doi: 10.1073/pnas.79.17.5240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lanzrein M., Schlegel A., Kempf C. Entry and uncoating of enveloped viruses. Biochem J. 1994 Sep 1;302(Pt 2):313–320. doi: 10.1042/bj3020313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Marcus P. I., Sekellick M. J., Johnson L. D., Lazzarini R. A. Cell killing by viruses. V. Transcribing defective interfering particles of vesicular stomatitis virus function as cell-killing particles. Virology. 1977 Oct 1;82(1):242–246. doi: 10.1016/0042-6822(77)90048-4. [DOI] [PubMed] [Google Scholar]
  15. Marsh M., Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–151. doi: 10.1016/S0065-3527(08)60583-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marsh M. The entry of enveloped viruses into cells by endocytosis. Biochem J. 1984 Feb 15;218(1):1–10. doi: 10.1042/bj2180001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mastromarino P., Conti C., Goldoni P., Hauttecoeur B., Orsi N. Characterization of membrane components of the erythrocyte involved in vesicular stomatitis virus attachment and fusion at acidic pH. J Gen Virol. 1987 Sep;68(Pt 9):2359–2369. doi: 10.1099/0022-1317-68-9-2359. [DOI] [PubMed] [Google Scholar]
  18. Matlin K. S., Reggio H., Helenius A., Simons K. Pathway of vesicular stomatitis virus entry leading to infection. J Mol Biol. 1982 Apr 15;156(3):609–631. doi: 10.1016/0022-2836(82)90269-8. [DOI] [PubMed] [Google Scholar]
  19. McAllister P. E., Wagner R. R. Differential inhibition of host protein synthesis in L cells infected with RNA - temperature-sensitive mutants of vesicular stomatitis virus. J Virol. 1976 May;18(2):550–558. doi: 10.1128/jvi.18.2.550-558.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McGowan J. J., Emerson S. U., Wagner R. R. The plus-strand leader RNA of VSV inhibits DNA-dependent transcription of adenovirus and SV40 genes in a soluble whole-cell extract. Cell. 1982 Feb;28(2):325–333. doi: 10.1016/0092-8674(82)90350-6. [DOI] [PubMed] [Google Scholar]
  21. Nicotera P., McConkey D. J., Jones D. P., Orrenius S. ATP stimulates Ca2+ uptake and increases the free Ca2+ concentration in isolated rat liver nuclei. Proc Natl Acad Sci U S A. 1989 Jan;86(2):453–457. doi: 10.1073/pnas.86.2.453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pinto L. H., Holsinger L. J., Lamb R. A. Influenza virus M2 protein has ion channel activity. Cell. 1992 May 1;69(3):517–528. doi: 10.1016/0092-8674(92)90452-i. [DOI] [PubMed] [Google Scholar]
  23. Schlegel R., Tralka T. S., Willingham M. C., Pastan I. Inhibition of VSV binding and infectivity by phosphatidylserine: is phosphatidylserine a VSV-binding site? Cell. 1983 Feb;32(2):639–646. doi: 10.1016/0092-8674(83)90483-x. [DOI] [PubMed] [Google Scholar]
  24. Schubert M., Lazzarini R. A. In vivo transcription of the 5'-terminal extracistronic region of vesicular stomatitis virus RNA. J Virol. 1981 Apr;38(1):256–262. doi: 10.1128/jvi.38.1.256-262.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Silva J. L., Luan P., Glaser M., Voss E. W., Weber G. Effects of hydrostatic pressure on a membrane-enveloped virus: high immunogenicity of the pressure-inactivated virus. J Virol. 1992 Apr;66(4):2111–2117. doi: 10.1128/jvi.66.4.2111-2117.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Silva J. L., Weber G. Pressure stability of proteins. Annu Rev Phys Chem. 1993;44:89–113. doi: 10.1146/annurev.pc.44.100193.000513. [DOI] [PubMed] [Google Scholar]
  27. Weck P. K., Carroll A. R., Shattuck D. M., Wagner R. R. Use of UV irradiation to identify the genetic information of vesicular stomatitis virus responsible for shutting off cellular RNA synthesis. J Virol. 1979 Jun;30(3):746–753. doi: 10.1128/jvi.30.3.746-753.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weck P. K., Wagner R. R. Inhibition of RNA synthesis in mouse myeloma cells infected with vesicular stomatitis virus. J Virol. 1978 Mar;25(3):770–780. doi: 10.1128/jvi.25.3.770-780.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weck P. K., Wagner R. R. Transcription of vesicular stomatitis virus is required to shut off cellular RNA synthesis. J Virol. 1979 Apr;30(1):410–413. doi: 10.1128/jvi.30.1.410-413.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Whitaker-Dowling P., Youngner J. S. Alteration of vesicular stomatitis virus L and NS proteins by uv irradiation: implications for the mechanism of host cell shut-off. Virology. 1988 May;164(1):171–175. doi: 10.1016/0042-6822(88)90633-2. [DOI] [PubMed] [Google Scholar]
  31. White J. M. Viral and cellular membrane fusion proteins. Annu Rev Physiol. 1990;52:675–697. doi: 10.1146/annurev.ph.52.030190.003331. [DOI] [PubMed] [Google Scholar]
  32. White J., Matlin K., Helenius A. Cell fusion by Semliki Forest, influenza, and vesicular stomatitis viruses. J Cell Biol. 1981 Jun;89(3):674–679. doi: 10.1083/jcb.89.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wu F. S., Lucas-Lenard J. M. Inhibition of ribonucleic acid accumulation in mouse L cells infected with vesicular stomatitis virus requires viral ribonucleic acid transcription. Biochemistry. 1980 Feb 19;19(4):804–810. doi: 10.1021/bi00545a029. [DOI] [PubMed] [Google Scholar]

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