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. 1992 Sep;66(9):5320–5328. doi: 10.1128/jvi.66.9.5320-5328.1992

A cis-acting DNA signal for encapsidation of simian virus 40.

A Oppenheim 1, Z Sandalon 1, A Peleg 1, O Shaul 1, S Nicolis 1, S Ottolenghi 1
PMCID: PMC289087  PMID: 1323699

Abstract

Encapsidation of simian virus 40 is a complex biological process involving DNA-protein and protein-protein interactions in the formation of a unique three-dimensional structure around the viral minichromosome. A pseudoviral system developed in our laboratory, in which the viral early and late gene products are supplied in trans (by helpers), was used to analyze the encapsidation process independent of viral gene expression. With this experimental system we have discovered a requirement for a specific DNA signal for encapsidation, ses (for simian virus 40 encapsidation signal).ses is present within a 200-bp DNA fragment, which includes, in addition to the viral origin of replication (ori), six GGGCGG repeats (GC boxes) and 26 bp of the enhancer element. Deletion of the GC boxes and the enhancer sequences almost abolished encapsidation, while DNA replication was only moderately decreased. The ability to encapsidate was not regained by reinserting a DNA fragment carrying ses in the sesdeleted plasmid 2 kbp away from the ori, suggesting that for encapsidation the two DNA elements have to be close to each other. These findings afford novel strategies for the investigation of viral encapsidation.

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Selected References

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

  1. Ambrose C., Blasquez V., Bina M. A block in initiation of simian virus 40 assembly results in the accumulation of minichromosomes containing an exposed regulatory region. Proc Natl Acad Sci U S A. 1986 May;83(10):3287–3291. doi: 10.1073/pnas.83.10.3287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderer F. A., Schlumberger H. D., Koch M. A., Frank H., Eggers H. J. Structure of simian virus 40. II. Symmetry and components of the virus particle. Virology. 1967 Jul;32(3):511–523. doi: 10.1016/0042-6822(67)90303-0. [DOI] [PubMed] [Google Scholar]
  3. Baker T. S., Drak J., Bina M. Reconstruction of the three-dimensional structure of simian virus 40 and visualization of the chromatin core. Proc Natl Acad Sci U S A. 1988 Jan;85(2):422–426. doi: 10.1073/pnas.85.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bargonetti J., Friedman P. N., Kern S. E., Vogelstein B., Prives C. Wild-type but not mutant p53 immunopurified proteins bind to sequences adjacent to the SV40 origin of replication. Cell. 1991 Jun 14;65(6):1083–1091. doi: 10.1016/0092-8674(91)90560-l. [DOI] [PubMed] [Google Scholar]
  5. Bergsma D. J., Olive D. M., Hartzell S. W., Subramanian K. N. Territorial limits and functional anatomy of the simian virus 40 replication origin. Proc Natl Acad Sci U S A. 1982 Jan;79(2):381–385. doi: 10.1073/pnas.79.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Buchanan R. L., Gralla J. D. Factor interactions at simian virus 40 GC-box promoter elements in intact nuclei. Mol Cell Biol. 1987 Apr;7(4):1554–1558. doi: 10.1128/mcb.7.4.1554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Chang X. B., Wilson J. H. Formation of deletions after initiation of simian virus 40 replication: influence of packaging limit of the capsid. J Virol. 1986 May;58(2):393–401. doi: 10.1128/jvi.58.2.393-401.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chen S. S., Hsu M. T. Evidence for variation of supercoil densities among simian virus 40 nucleoprotein complexes and for higher supercoil density in replicating complexes. J Virol. 1984 Jul;51(1):14–19. doi: 10.1128/jvi.51.1.14-19.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Coca-Prados M., Yu H. Y., Hsu M. T. Intracellular forms of simian virus 40 nucleoprotein complexes. IV. Micrococcal nuclease digestion. J Virol. 1982 Nov;44(2):603–609. doi: 10.1128/jvi.44.2.603-609.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dynan W. S., Tjian R. The promoter-specific transcription factor Sp1 binds to upstream sequences in the SV40 early promoter. Cell. 1983 Nov;35(1):79–87. doi: 10.1016/0092-8674(83)90210-6. [DOI] [PubMed] [Google Scholar]
  12. Garber E. A., Seidman M. M., Levine A. J. Intracellular SV40 nucleoprotein complexes: synthesis to encapsidation. Virology. 1980 Dec;107(2):389–401. doi: 10.1016/0042-6822(80)90306-2. [DOI] [PubMed] [Google Scholar]
  13. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  14. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Guo Z. S., Gutierrez C., Heine U., Sogo J. M., Depamphilis M. L. Origin auxiliary sequences can facilitate initiation of simian virus 40 DNA replication in vitro as they do in vivo. Mol Cell Biol. 1989 Sep;9(9):3593–3602. doi: 10.1128/mcb.9.9.3593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hartman J. R., Nayak D. P., Fareed G. C. Human influenza virus hemagglutinin is expressed in monkey cells using simian virus 40 vectors. Proc Natl Acad Sci U S A. 1982 Jan;79(2):233–237. doi: 10.1073/pnas.79.2.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hearing P., Samulski R. J., Wishart W. L., Shenk T. Identification of a repeated sequence element required for efficient encapsidation of the adenovirus type 5 chromosome. J Virol. 1987 Aug;61(8):2555–2558. doi: 10.1128/jvi.61.8.2555-2558.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  19. Jakobovits E. B., Bratosin S., Aloni Y. A nucleosome-free region in SV40 minichromosomes. Nature. 1980 May 22;285(5762):263–265. doi: 10.1038/285263a0. [DOI] [PubMed] [Google Scholar]
  20. Khoury G., Lai C. J. Preparation of simian virus 40 and its DNA. Methods Enzymol. 1979;58:404–412. doi: 10.1016/s0076-6879(79)58155-5. [DOI] [PubMed] [Google Scholar]
  21. Liddington R. C., Yan Y., Moulai J., Sahli R., Benjamin T. L., Harrison S. C. Structure of simian virus 40 at 3.8-A resolution. Nature. 1991 Nov 28;354(6351):278–284. doi: 10.1038/354278a0. [DOI] [PubMed] [Google Scholar]
  22. Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
  23. Martin R. G. On the nucleoprotein core of simian virus 40. Virology. 1977 Dec;83(2):433–437. doi: 10.1016/0042-6822(77)90190-8. [DOI] [PubMed] [Google Scholar]
  24. McCutchan J. H., Pagano J. S. Enchancement of the infectivity of simian virus 40 deoxyribonucleic acid with diethylaminoethyl-dextran. J Natl Cancer Inst. 1968 Aug;41(2):351–357. [PubMed] [Google Scholar]
  25. 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]
  26. Oppenheim A., Peleg A., Fibach E., Rachmilewitz E. A. Efficient introduction of plasmid DNA into human hemopoietic cells by encapsidation in simian virus 40 pseudovirions. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6925–6929. doi: 10.1073/pnas.83.18.6925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oppenheim A., Peleg A. Helpers for efficient encapsidation of SV40 pseudovirions. Gene. 1989 Apr 15;77(1):79–86. doi: 10.1016/0378-1119(89)90361-2. [DOI] [PubMed] [Google Scholar]
  28. Oppenheim A., Peleg A., Rachmilewitz E. A. Efficient introduction and transient expression of exogenous genes in human hemopoietic cells. Ann N Y Acad Sci. 1987;511:418–427. doi: 10.1111/j.1749-6632.1987.tb36271.x. [DOI] [PubMed] [Google Scholar]
  29. Rao B. S., Martin R. G. DpnI assay for DNA replication in animal cells: enzyme-resistant material can result from factors not related to replication. Nucleic Acids Res. 1988 May 11;16(9):4171–4171. doi: 10.1093/nar/16.9.4171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  32. Salunke D. M., Caspar D. L., Garcea R. L. Self-assembly of purified polyomavirus capsid protein VP1. Cell. 1986 Sep 12;46(6):895–904. doi: 10.1016/0092-8674(86)90071-1. [DOI] [PubMed] [Google Scholar]
  33. Saragosti S., Moyne G., Yaniv M. Absence of nucleosomes in a fraction of SV40 chromatin between the origin of replication and the region coding for the late leader RNA. Cell. 1980 May;20(1):65–73. doi: 10.1016/0092-8674(80)90235-4. [DOI] [PubMed] [Google Scholar]
  34. Soussi T. DNA-binding properties of the major structural protein of simian virus 40. J Virol. 1986 Sep;59(3):740–742. doi: 10.1128/jvi.59.3.740-742.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  36. Stillman B., Gerard R. D., Guggenheimer R. A., Gluzman Y. T antigen and template requirements for SV40 DNA replication in vitro. EMBO J. 1985 Nov;4(11):2933–2939. doi: 10.1002/j.1460-2075.1985.tb04026.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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