Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1996 Feb;70(2):1317–1322. doi: 10.1128/jvi.70.2.1317-1322.1996

Analysis of a nuclear localization signal of simian virus 40 major capsid protein Vp1.

N Ishii 1, N Minami 1, E Y Chen 1, A L Medina 1, M M Chico 1, H Kasamatsu 1
PMCID: PMC189950  PMID: 8551602

Abstract

The nuclear localization signal of the major structural protein, Vp1, of simian virus 40 was further defined by mutagenesis. The targeting activity was examined in cells microinjected with SV-Vp1 variant viral DNAs bearing either an initiation codon mutation of the agnoprotein or mutations in the Vp1 coding sequence or microinjected with pSG5-Vp1 and pSG5-Vp1 mutant DNAs in which Vp1 or mutant Vp1 is expressed from simian virus 40 early promoter. The Vp1 nuclear localization signal functioned autonomously without agno-protein once the Vp1 protein was synthesized in the cytoplasm. The targeting activity was localized to the amino-terminal 19 residues. While replacement of cysteine 10 with glycine, alanine, or serine did not affect the activity, replacement of arginine 6 with glycine caused the cytoplasmic phenotype. When multiple mutations were introduced among residue 5, 6, 7, 16, 17, or 19, the targeting activity was found to reside in two clusters of basic residues, a cluster of lysine 5, arginine 6, and lysine 7 and a cluster of lysine 16, lysine 17, and lysine 19. The clusters are independently important for nuclear localization activity.

Full Text

The Full Text of this article is available as a PDF (423.9 KB).

Selected References

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

  1. Alwine J. C. Evidence for simian virus 40 late transcriptional control: mixed infections of wild-type simian virus 40 and a late leader deletion mutant exhibit trans effects on late viral RNA synthesis. J Virol. 1982 Jun;42(3):798–803. doi: 10.1128/jvi.42.3.798-803.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barkan A., Mertz J. E. DNA sequence analysis of simian virus 40 mutants with deletions mapping in the leader region of the late viral mRNA's: mutants with deletions similar in size and position exhibit varied phenotypes. J Virol. 1981 Feb;37(2):730–737. doi: 10.1128/jvi.37.2.730-737.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carswell S., Alwine J. C. Simian virus 40 agnoprotein facilitates perinuclear-nuclear localization of VP1, the major capsid protein. J Virol. 1986 Dec;60(3):1055–1061. doi: 10.1128/jvi.60.3.1055-1061.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang D., Haynes J. I., 2nd, Brady J. N., Consigli R. A. Identification of a nuclear localization sequence in the polyomavirus capsid protein VP2. Virology. 1992 Dec;191(2):978–983. doi: 10.1016/0042-6822(92)90276-u. [DOI] [PubMed] [Google Scholar]
  5. Chang D., Haynes J. I., 2nd, Brady J. N., Consigli R. A. The use of additive and subtractive approaches to examine the nuclear localization sequence of the polyomavirus major capsid protein VP1. Virology. 1992 Aug;189(2):821–827. doi: 10.1016/0042-6822(92)90615-v. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Dingwall C., Laskey R. A. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. doi: 10.1016/0968-0004(91)90184-w. [DOI] [PubMed] [Google Scholar]
  9. Garcia-Bustos J., Heitman J., Hall M. N. Nuclear protein localization. Biochim Biophys Acta. 1991 Mar 7;1071(1):83–101. doi: 10.1016/0304-4157(91)90013-m. [DOI] [PubMed] [Google Scholar]
  10. Gharakhanian E., Takahashi J., Kasamatsu H. The carboxyl 35 amino acids of SV40 Vp3 are essential for its nuclear accumulation. Virology. 1987 Apr;157(2):440–448. doi: 10.1016/0042-6822(87)90286-8. [DOI] [PubMed] [Google Scholar]
  11. Ghosh P. K., Piatak M., Mertz J. E., Weissman S. M., Lebowitz P. Altered utilization of splice sites and 5' termini in late RNAs produced by leader region mutants of simian virus 40. J Virol. 1982 Nov;44(2):610–624. doi: 10.1128/jvi.44.2.610-624.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hay N., Aloni Y. Attenuation of late simian virus 40 mRNA synthesis is enhanced by the agnoprotein and is temporally regulated in isolated nuclear systems. Mol Cell Biol. 1985 Jun;5(6):1327–1334. doi: 10.1128/mcb.5.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ishii N., Nakanishi A., Yamada M., Macalalad M. H., Kasamatsu H. Functional complementation of nuclear targeting-defective mutants of simian virus 40 structural proteins. J Virol. 1994 Dec;68(12):8209–8216. doi: 10.1128/jvi.68.12.8209-8216.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jay G., Nomura S., Anderson C. W., Khoury G. Identification of the SV40 agnogene product: a DNA binding protein. Nature. 1981 May 28;291(5813):346–349. doi: 10.1038/291346a0. [DOI] [PubMed] [Google Scholar]
  15. Kleinschmidt J. A., Seiter A. Identification of domains involved in nuclear uptake and histone binding of protein N1 of Xenopus laevis. EMBO J. 1988 Jun;7(6):1605–1614. doi: 10.1002/j.1460-2075.1988.tb02986.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Margolskee R. F., Nathans D. Suppression of a VP1 mutant of simian virus 40 by missense mutations in serine codons of the viral agnogene. J Virol. 1983 Nov;48(2):405–409. doi: 10.1128/jvi.48.2.405-409.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Moreland R. B., Garcea R. L. Characterization of a nuclear localization sequence in the polyomavirus capsid protein VP1. Virology. 1991 Nov;185(1):513–518. doi: 10.1016/0042-6822(91)90811-o. [DOI] [PubMed] [Google Scholar]
  18. Ng S. C., Mertz J. E., Sanden-Will S., Bina M. Simian virus 40 maturation in cells harboring mutants deleted in the agnogene. J Biol Chem. 1985 Jan 25;260(2):1127–1132. [PubMed] [Google Scholar]
  19. Nomura S., Khoury G., Jay G. Subcellular localization of the simian virus 40 agnoprotein. J Virol. 1983 Jan;45(1):428–433. doi: 10.1128/jvi.45.1.428-433.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Resnick J., Shenk T. Simian virus 40 agnoprotein facilitates normal nuclear location of the major capsid polypeptide and cell-to-cell spread of virus. J Virol. 1986 Dec;60(3):1098–1106. doi: 10.1128/jvi.60.3.1098-1106.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Richardson W. D., Roberts B. L., Smith A. E. Nuclear location signals in polyoma virus large-T. Cell. 1986 Jan 17;44(1):77–85. doi: 10.1016/0092-8674(86)90486-1. [DOI] [PubMed] [Google Scholar]
  22. Robbins J., Dilworth S. M., Laskey R. A., Dingwall C. Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell. 1991 Feb 8;64(3):615–623. doi: 10.1016/0092-8674(91)90245-t. [DOI] [PubMed] [Google Scholar]
  23. Sedman S. A., Good P. J., Mertz J. E. Leader-encoded open reading frames modulate both the absolute and relative rates of synthesis of the virion proteins of simian virus 40. J Virol. 1989 Sep;63(9):3884–3893. doi: 10.1128/jvi.63.9.3884-3893.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Underwood M. R., Fried H. M. Characterization of nuclear localizing sequences derived from yeast ribosomal protein L29. EMBO J. 1990 Jan;9(1):91–99. doi: 10.1002/j.1460-2075.1990.tb08084.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Wychowski C., Benichou D., Girard M. The intranuclear location of simian virus 40 polypeptides VP2 and VP3 depends on a specific amino acid sequence. J Virol. 1987 Dec;61(12):3862–3869. doi: 10.1128/jvi.61.12.3862-3869.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES