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
. 1991 Aug 15;88(16):7333–7337. doi: 10.1073/pnas.88.16.7333

Import of simian virus 40 virions through nuclear pore complexes.

J Clever 1, M Yamada 1, H Kasamatsu 1
PMCID: PMC52289  PMID: 1651501

Abstract

How the DNA tumor virus, simian virus 40, reaches the nucleus is unknown. In this report we have tested the affinity of simian virus 40 toward the nucleus by microinjecting virion particles into the cytoplasm under conditions in which cell-surface-mediated viral infection was blocked. Subcellular localization of viral structural proteins Vp1, Vp2, and Vp3, large tumor antigen, and virion particles was followed immunocytochemically and ultrastructurally. Both virion particles and viral structural proteins localized in the nucleus within 1-2 hr after cytoplasmic injection and subsequently expressed large tumor antigen, which was detected in the nucleus as early as 3 hr after cytoplasmic injection. Vp1 and large tumor antigen nuclear accumulation, as well as virion nuclear entry, were blocked by wheat germ agglutinin and an anti-nucleoporin monoclonal antibody, mAb 414. Virion particles were visualized in the vicinity of nuclear pores and in the cytoplasm with this agent. We conclude that virion particles are karyophilic and enter through nuclear pores. This study suggests that virion structural proteins facilitate virion import into the nucleus and viral gene expression.

Full text

PDF
7333

Images in this article

7334Image
on p.7334
7335Image
on p.7335
7335Image
on p.7335
7335Image
on p.7335
7335Image
on p.7335
7336Image
on p.7336

Selected References

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

  1. Akey C. W., Goldfarb D. S. Protein import through the nuclear pore complex is a multistep process. J Cell Biol. 1989 Sep;109(3):971–982. doi: 10.1083/jcb.109.3.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Akey C. W. Visualization of transport-related configurations of the nuclear pore transporter. Biophys J. 1990 Aug;58(2):341–355. doi: 10.1016/S0006-3495(90)82381-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barbanti-Brodano G., Swetly P., Koprowski H. Early events in the infection of permissive cells with simian virus 40: adsorption, penetration, and uncoating. J Virol. 1970 Jul;6(1):78–86. doi: 10.1128/jvi.6.1.78-86.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Black P. H., Rowe W. P. Induction of SV40 T antigen with SV40 DNA. Virology. 1965 Nov;27(3):436–439. doi: 10.1016/0042-6822(65)90127-3. [DOI] [PubMed] [Google Scholar]
  5. Chelsky D., Ralph R., Jonak G. Sequence requirements for synthetic peptide-mediated translocation to the nucleus. Mol Cell Biol. 1989 Jun;9(6):2487–2492. doi: 10.1128/mcb.9.6.2487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clever J., Kasamatsu H. Simian virus 40 Vp2/3 small structural proteins harbor their own nuclear transport signal. Virology. 1991 Mar;181(1):78–90. doi: 10.1016/0042-6822(91)90472-n. [DOI] [PubMed] [Google Scholar]
  7. Davis L. I., Blobel G. Identification and characterization of a nuclear pore complex protein. Cell. 1986 Jun 6;45(5):699–709. doi: 10.1016/0092-8674(86)90784-1. [DOI] [PubMed] [Google Scholar]
  8. Dworetzky S. I., Lanford R. E., Feldherr C. M. The effects of variations in the number and sequence of targeting signals on nuclear uptake. J Cell Biol. 1988 Oct;107(4):1279–1287. doi: 10.1083/jcb.107.4.1279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Featherstone C., Darby M. K., Gerace L. A monoclonal antibody against the nuclear pore complex inhibits nucleocytoplasmic transport of protein and RNA in vivo. J Cell Biol. 1988 Oct;107(4):1289–1297. doi: 10.1083/jcb.107.4.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feldherr C. M., Kallenbach E., Schultz N. Movement of a karyophilic protein through the nuclear pores of oocytes. J Cell Biol. 1984 Dec;99(6):2216–2222. doi: 10.1083/jcb.99.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Filson A. J., Lewis A., Blobel G., Fisher P. A. Monoclonal antibodies prepared against the major Drosophila nuclear Matrix-pore complex-lamina glycoprotein bind specifically to the nuclear envelope in situ. J Biol Chem. 1985 Mar 10;260(5):3164–3172. [PubMed] [Google Scholar]
  12. Finlay D. R., Newmeyer D. D., Price T. M., Forbes D. J. Inhibition of in vitro nuclear transport by a lectin that binds to nuclear pores. J Cell Biol. 1987 Feb;104(2):189–200. doi: 10.1083/jcb.104.2.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gerace L., Ottaviano Y., Kondor-Koch C. Identification of a major polypeptide of the nuclear pore complex. J Cell Biol. 1982 Dec;95(3):826–837. doi: 10.1083/jcb.95.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gharakhanian E., Kasamatsu H. Two independent signals, a nuclear localization signal and a Vp1-interactive signal, reside within the carboxy-35 amino acids of SV40 Vp3. Virology. 1990 Sep;178(1):62–71. doi: 10.1016/0042-6822(90)90379-6. [DOI] [PubMed] [Google Scholar]
  15. Greber U. F., Senior A., Gerace L. A major glycoprotein of the nuclear pore complex is a membrane-spanning polypeptide with a large lumenal domain and a small cytoplasmic tail. EMBO J. 1990 May;9(5):1495–1502. doi: 10.1002/j.1460-2075.1990.tb08267.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Griffith G. R., Marriott S. J., Rintoul D. A., Consigli R. A. Early events in polyomavirus infection: fusion of monopinocytotic vesicles containing virions with mouse kidney cell nuclei. Virus Res. 1988 Apr;10(1):41–51. doi: 10.1016/0168-1702(88)90056-1. [DOI] [PubMed] [Google Scholar]
  17. Hummeler K., Tomassini N., Sokol F. Morphological aspects of the uptake of simian virus 40 by permissive cells. J Virol. 1970 Jul;6(1):87–93. doi: 10.1128/jvi.6.1.87-93.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kartenbeck J., Stukenbrok H., Helenius A. Endocytosis of simian virus 40 into the endoplasmic reticulum. J Cell Biol. 1989 Dec;109(6 Pt 1):2721–2729. doi: 10.1083/jcb.109.6.2721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kasamatsu H., Nehorayan A. Intracellular localization of viral polypeptides during simian virus 40 infection. J Virol. 1979 Nov;32(2):648–660. doi: 10.1128/jvi.32.2.648-660.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kasamatsu H., Wu M. Protein-SV40 DNA complex stable in high salt and sodium dodecyl sulfate. Biochem Biophys Res Commun. 1976 Feb 9;68(3):927–936. doi: 10.1016/0006-291x(76)91234-1. [DOI] [PubMed] [Google Scholar]
  21. Lin W., Hata T., Kasamatsu H. Subcellular distribution of viral structural proteins during simian virus 40 infection. J Virol. 1984 May;50(2):363–371. doi: 10.1128/jvi.50.2.363-371.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin W., Shurgot J. L., Kasamatsu H. The synthesis and transport of SV40 structural proteins. Virology. 1986 Oct 15;154(1):108–120. doi: 10.1016/0042-6822(86)90434-4. [DOI] [PubMed] [Google Scholar]
  23. Mackay R. L., Consigli R. A. Early events in polyoma virus infection: attachment, penetration, and nuclear entry. J Virol. 1976 Aug;19(2):620–636. doi: 10.1128/jvi.19.2.620-636.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maraldi N. M., Barbanti-Brodano G., Portolani M., La Placa M. Ultrastructural aspects of BK virus uptake and replication in human fibroblasts. J Gen Virol. 1975 Apr;27(1):71–80. doi: 10.1099/0022-1317-27-1-71. [DOI] [PubMed] [Google Scholar]
  25. Maul G. G., Rovera G., Vorbrodt A., Abramczuk J. Membrane fusion as a mechanism of simian virus 40 entry into different cellular compartments. J Virol. 1978 Dec;28(3):936–944. doi: 10.1128/jvi.28.3.936-944.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nishimura T., Kawai N., Kawai M., Notake K., Ichihara I. Fusion of SV40-induced endocytotic vacuoles with the nuclear membrane. Cell Struct Funct. 1986 Jun;11(2):135–141. doi: 10.1247/csf.11.135. [DOI] [PubMed] [Google Scholar]
  27. RAPP F., KITAHARA T., BUTEL J. S., MELNICK J. L. SYNTHESIS OF SV40 TUMOR ANTIGEN DURING REPLICATION OF SIMIAN PAPOVAVIRUS (SV40). Proc Natl Acad Sci U S A. 1964 Nov;52:1138–1142. doi: 10.1073/pnas.52.5.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wozniak R. W., Bartnik E., Blobel G. Primary structure analysis of an integral membrane glycoprotein of the nuclear pore. J Cell Biol. 1989 Jun;108(6):2083–2092. doi: 10.1083/jcb.108.6.2083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wychowski C., Benichou D., Girard M. A domain of SV40 capsid polypeptide VP1 that specifies migration into the cell nucleus. EMBO J. 1986 Oct;5(10):2569–2576. doi: 10.1002/j.1460-2075.1986.tb04536.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamada M., Moritoh C., Kawaguchi M., Okigaki T. Growth, morphology, function, and morphogenetic properties of rat renal glomerular epithelial cells in vitro: effects of retinyl acetate. Eur J Cell Biol. 1989 Aug;49(2):252–258. [PubMed] [Google Scholar]
  31. Yoneda Y., Imamoto-Sonobe N., Yamaizumi M., Uchida T. Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells. Exp Cell Res. 1987 Dec;173(2):586–595. doi: 10.1016/0014-4827(87)90297-7. [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