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. 1982;1(1):9–14. doi: 10.1002/j.1460-2075.1982.tb01116.x

Transcription of Simian Virus 40 chromosomes in an extract of HeLa cells.

P Beard, K Nyfeler
PMCID: PMC552987  PMID: 6325152

Abstract

Simian Virus 40 (SV40) chromosomes were incubated with a concentrated extract of HeLa cells containing RNA polymerase II and other factors involved in transcription. SV40-specific RNA was synthesized. In the absence of HeLa cell extract the synthesis of labeled RNA by endogenous RNA polymerase in the chromosome preparations amounted to less than one tenth of that when the HeLa cell extract was present. Incubation with the HeLa extract increased the amount of Sarkosyl-resistant (i.e., transcribing) RNA polymerase on the SV40 chromosomes. When restriction endonucleases able to cut SV40 DNA were added to transcription reactions containing SV40 chromosomes and the HeLa cell extract, RNAs of discrete length were produced. The RNAs were identified as correctly initiated run-off transcripts of the early and late genes. Most of the RNA synthesized by the HeLa extract from SV40 chromosomes was from the region of the late genes, whereas transcription of purified SV40 DNA was from both the early and the late regions, with the early transcripts predominating.

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Selected References

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

  1. Alwine J. C., Reed S. I., Stark G. R. Characterization of the autoregulation of simian virus 40 gene A. J Virol. 1977 Oct;24(1):22–27. doi: 10.1128/jvi.24.1.22-27.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beard P., Kaneko M., Cerutti P. N-Acetoxy-acetylaminofluorene reacts preferentially with a control region of intracellular SV40 chromosome. Nature. 1981 May 7;291(5810):84–85. doi: 10.1038/291084a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crémisi C., Pignatti P. F., Yaniv M. Random location and absence of movement of the nucleosomes on SV 40 nucleoprotein complex isolated from infected cells. Biochem Biophys Res Commun. 1976 Dec 6;73(3):548–554. doi: 10.1016/0006-291x(76)90845-7. [DOI] [PubMed] [Google Scholar]
  6. Dubochet J., Ducommun M., Zollinger M., Kellenberger E. A new preparation method for dark-field electron microscopy of biomacromolecules. J Ultrastruct Res. 1971 Apr;35(1):147–167. doi: 10.1016/s0022-5320(71)80148-x. [DOI] [PubMed] [Google Scholar]
  7. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  8. Felsenfeld G. Chromatin. Nature. 1978 Jan 12;271(5641):115–122. doi: 10.1038/271115a0. [DOI] [PubMed] [Google Scholar]
  9. Gariglio P., Buss J., Green M. H. Sarkosyl activation of RNA polymerase activity in mitotic mouse cells. FEBS Lett. 1974 Aug 30;44(3):330–333. doi: 10.1016/0014-5793(74)81170-1. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Germond J. E., Hirt B., Oudet P., Gross-Bellark M., Chambon P. Folding of the DNA double helix in chromatin-like structures from simian virus 40. Proc Natl Acad Sci U S A. 1975 May;72(5):1843–1847. doi: 10.1073/pnas.72.5.1843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ghosh P. K., Reddy V. B., Swinscoe J., Choudary P. V., Lebowitz P., Weissman S. M. The 5'-terminal leader sequence of late 16 S mRNA from cells infected with simian virus 40. J Biol Chem. 1978 May 25;253(10):3643–3647. [PubMed] [Google Scholar]
  13. Green M. R., Lebovitz R. M., Roeder R. G. Expression of the autonomous parvovirus H1 genome: evidence for a single transcriptional unit and multiple spliced polyadenylated transcripts. Cell. 1979 Aug;17(4):967–977. doi: 10.1016/0092-8674(79)90336-2. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Handa H., Kaufman R. J., Manley J., Gefter M., Sharp P. A. Transcription of Simian virus 40 DNA in a HeLa whole cell extract. J Biol Chem. 1981 Jan 10;256(1):478–482. [PubMed] [Google Scholar]
  16. 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]
  17. Keller W., Müller U., Eicken I., Wendel I., Zentgraf H. Biochemical and ultrastructural analysis of SV40 chromatin. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):227–244. doi: 10.1101/sqb.1978.042.01.025. [DOI] [PubMed] [Google Scholar]
  18. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Llopis R., Perrin F., Bellard F., Gariglio P. Quantitation of transcribing native simian virus 40 minichromosomes extracted from CV1 cells late in infection. J Virol. 1981 Apr;38(1):82–90. doi: 10.1128/jvi.38.1.82-90.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Mann K., Hunter T. Association of simian virus 40 T antigen with simian virus 40 nucleoprotein complexes. J Virol. 1979 Jan;29(1):232–241. doi: 10.1128/jvi.29.1.232-241.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murray K., Murray N. E. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli. J Mol Biol. 1975 Nov 5;98(3):551–564. doi: 10.1016/s0022-2836(75)80086-6. [DOI] [PubMed] [Google Scholar]
  24. Proudfoot N. J., Shander M. H., Manley J. L., Gefter M. L., Maniatis T. Structure and in vitro transcription of human globin genes. Science. 1980 Sep 19;209(4463):1329–1336. doi: 10.1126/science.6158093. [DOI] [PubMed] [Google Scholar]
  25. Reiser J., Renart J., Crawford L. V., Stark G. R. Specific association of simian virus 40 tumor antigen with simian virus 40 chromatin. J Virol. 1980 Jan;33(1):78–87. doi: 10.1128/jvi.33.1.78-87.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Scott W. A., Wigmore D. J. Sites in simian virus 40 chromatin which are preferentially cleaved by endonucleases. Cell. 1978 Dec;15(4):1511–1518. doi: 10.1016/0092-8674(78)90073-9. [DOI] [PubMed] [Google Scholar]
  28. Su R. T., DePamphilis M. L. In vitro replication of simian virus 40 DNA in a nucleoprotein complex. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3466–3470. doi: 10.1073/pnas.73.10.3466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. 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]

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