Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1981 Apr;38(1):82–90. doi: 10.1128/jvi.38.1.82-90.1981

Quantitation of transcribing native simian virus 40 minichromosomes extracted from CV1 cells late in infection.

R Llopis, F Perrin, F Bellard, P Gariglio
PMCID: PMC171128  PMID: 6264116

Abstract

Simian virus 40 transcriptional complexes could be extracted from CV1 cells late in infection and separated from the bulk of inactive viral chromatin. Sucrose gradient sedimentation, cesium sulfate equilibrium density gradient centrifugation, and electron microscopy indicated that the viral transcriptional complexes corresponded to at least 0.5 to 1% of the viral minichromosomes extracted, about 5,000 copies per cell.

Full text

PDF
82

Images in this article

87Image
on p.87

Selected References

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

  1. Aloni Y., Bratosin S., Dhar R., Laub O., Horowitz M., Khoury G. Splicing of SV40 mRNAs: a novel mechanism for the regulation of gene expression in animal cells. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):559–570. doi: 10.1101/sqb.1978.042.01.058. [DOI] [PubMed] [Google Scholar]
  2. Bellard M., Oudet P., Germond J. E., Chambon P. Subunit structure of simian-virus-40 minichromosome. Eur J Biochem. 1976 Nov 15;70(2):543–553. doi: 10.1111/j.1432-1033.1976.tb11046.x. [DOI] [PubMed] [Google Scholar]
  3. Birkenmeier E. H., Radonovich M. F., Shani M., Salzman N. P. The SV40 DNA template for transcription of late mRNA in viral nucleoprotein complexes. Cell. 1977 Jul;11(3):495–504. doi: 10.1016/0092-8674(77)90067-8. [DOI] [PubMed] [Google Scholar]
  4. Bloom K. S., Anderson J. N. Fractionation of hen oviduct chromatin into transcriptionally active and inactive regions after selective micrococcal nuclease digestion. Cell. 1978 Sep;15(1):141–150. doi: 10.1016/0092-8674(78)90090-9. [DOI] [PubMed] [Google Scholar]
  5. Britten R. J., Davidson E. H. Repetitive and non-repetitive DNA sequences and a speculation on the origins of evolutionary novelty. Q Rev Biol. 1971 Jun;46(2):111–138. doi: 10.1086/406830. [DOI] [PubMed] [Google Scholar]
  6. Chambon P. Summary: the molecular biology of the eukaryotic genome is coming of age. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):1209–1234. doi: 10.1101/sqb.1978.042.01.122. [DOI] [PubMed] [Google Scholar]
  7. Cox R. F. Transcription of high-molecular-weight RNA from hen-oviduct chromatin by bacterial and endogenous form-B RNA polymerases. Eur J Biochem. 1973 Nov 1;39(1):49–61. doi: 10.1111/j.1432-1033.1973.tb03102.x. [DOI] [PubMed] [Google Scholar]
  8. Fernandez-Munoz R., Coca-Prados M., Hsu M. T. Intracellular forms of simian virus 40 nucleoprotein complexes. I. Methods of isolation and characterization in CV-1 cells. J Virol. 1979 Feb;29(2):612–623. doi: 10.1128/jvi.29.2.612-623.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  10. Garber E. A., Seidman M. M., Levine A. J. The detection and characterization of multiple forms of SV40 nucleoprotein complexes. Virology. 1978 Oct 15;90(2):305–316. doi: 10.1016/0042-6822(78)90315-x. [DOI] [PubMed] [Google Scholar]
  11. Gariglio P., Llopis R., Oudet P., Chambon P. The template of the isolated native simian virus 40 transcriptional complexes is a minichromosome. J Mol Biol. 1979 Jun 15;131(1):75–105. doi: 10.1016/0022-2836(79)90302-4. [DOI] [PubMed] [Google Scholar]
  12. Gariglio P., Mousset S. Characterization of a soluble simian-virus-40 transcription complex. Eur J Biochem. 1977 Jun 15;76(2):583–590. doi: 10.1111/j.1432-1033.1977.tb11628.x. [DOI] [PubMed] [Google Scholar]
  13. Gariglio P., Mousset S. Isolation and partial characterization of a nuclear RNA polymerase - SV40 DNA complex. FEBS Lett. 1975 Aug 1;56(1):149–155. doi: 10.1016/0014-5793(75)80130-x. [DOI] [PubMed] [Google Scholar]
  14. Gottesfeld J. M., Butler P. J. Structure of transcriptionally-active chromatin subunits. Nucleic Acids Res. 1977 Sep;4(9):3155–3173. doi: 10.1093/nar/4.9.3155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Green M. H., Brooks T. L. Isolation of two forms of SV40 nucleoprotein containing RNA polymerase from infected monkey cells. Virology. 1976 Jul 1;72(1):110–120. doi: 10.1016/0042-6822(76)90316-0. [DOI] [PubMed] [Google Scholar]
  16. Green M. H., Miller H. I., Hendler S. Isolation of a polyoma-nucleoprotein complex from infected mouse-cell cultures. Proc Natl Acad Sci U S A. 1971 May;68(5):1032–1036. doi: 10.1073/pnas.68.5.1032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Griffith J. D. Chromatin structure: deduced from a minichromosome. Science. 1975 Mar 28;187(4182):1202–1203. doi: 10.1126/science.187.4182.1202. [DOI] [PubMed] [Google Scholar]
  18. Hendrick D., Tolstoshev P., Randlett D. Enrichment for the globin coding region in a chromatin fraction from chick reticulocytes by endonuclease digestion. Gene. 1977;2(3-4):147–158. doi: 10.1016/0378-1119(77)90014-2. [DOI] [PubMed] [Google Scholar]
  19. Jakobovits E. B., Aloni Y. Isolation and characterization of various forms of simian virus 40 DNA-protein complexes. Virology. 1980 Apr 15;102(1):107–118. doi: 10.1016/0042-6822(80)90074-4. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Laub O., Bratosin S., Horowitz M., Aloni Y. The initiation of transcription of SV40 DNA at late time after infection. Virology. 1979 Jan 30;92(2):310–323. doi: 10.1016/0042-6822(79)90136-3. [DOI] [PubMed] [Google Scholar]
  22. Laub O., Jakobovits E. B., Aloni Y. 5,6-dichloro-1-beta-ribofuranosylbenzimidazole enhances premature termination of late transcription of simian virus 40 DNA. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3297–3301. doi: 10.1073/pnas.77.6.3297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Llopis R., Stark G. R. Two deletions within genes for simian virus 40 structural proteins VP2 and VP3 lead to formation of abnormal transcriptional complexes. J Virol. 1981 Apr;38(1):91–103. doi: 10.1128/jvi.38.1.91-103.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Meinke W., Hall M. R., Goldstein D. A. Proteins in intracellular simian virus 40 nucleoportein complexes: comparison with simian virus 40 core proteins. J Virol. 1975 Mar;15(3):439–448. doi: 10.1128/jvi.15.3.439-448.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Müller U., Zentgraf H., Eicken I., Keller W. Higher order structure of simian virus 40 chromatin. Science. 1978 Aug 4;201(4354):406–415. doi: 10.1126/science.208155. [DOI] [PubMed] [Google Scholar]
  27. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Shani M., Birkenmeier E., May E., Salzman N. P. Properties of simian virus 40 transcriptional intermediates isolated from nuclei of permissive cells. J Virol. 1977 Jul;23(1):20–28. doi: 10.1128/jvi.23.1.20-28.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tata J. R., Baker B. Enzymatic fractionation of nuclei: polynucleosomes and RNA polymerase II as endogenous transcriptional complexes. J Mol Biol. 1978 Jan 25;118(3):249–272. doi: 10.1016/0022-2836(78)90227-9. [DOI] [PubMed] [Google Scholar]
  31. Tjian R. The binding site on SV40 DNA for a T antigen-related protein. Cell. 1978 Jan;13(1):165–179. doi: 10.1016/0092-8674(78)90147-2. [DOI] [PubMed] [Google Scholar]
  32. Varshavsky A. J., Bakayev V. V., Nedospasov S. A., Georgiev G. P. On the structure of eukaryotic, prokaryotic, and viral chromatin. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):457–473. doi: 10.1101/sqb.1978.042.01.049. [DOI] [PubMed] [Google Scholar]
  33. Varshavsky A. J., Sundin O. H., Bohn M. J. SV40 viral minichromosome: preferential exposure of the origin of replication as probed by restriction endonucleases. Nucleic Acids Res. 1978 Oct;5(10):3469–3477. doi: 10.1093/nar/5.10.3469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Varshavsky A. J., Sundin O., Bohn M. A stretch of "late" SV40 viral DNA about 400 bp long which includes the origin of replication is specifically exposed in SV40 minichromosomes. Cell. 1979 Feb;16(2):453–466. doi: 10.1016/0092-8674(79)90021-7. [DOI] [PubMed] [Google Scholar]
  35. Wasylyk B., Kédinger C., Corden J., Brison O., Chambon P. Specific in vitro initiation of transcription on conalbumin and ovalbumin genes and comparison with adenovirus-2 early and late genes. Nature. 1980 Jun 5;285(5764):367–373. doi: 10.1038/285367a0. [DOI] [PubMed] [Google Scholar]
  36. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES