Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1977 Jul;23(1):20–28. doi: 10.1128/jvi.23.1.20-28.1977

Properties of simian virus 40 transcriptional intermediates isolated from nuclei of permissive cells.

M Shani, E Birkenmeier, E May, N P Salzman
PMCID: PMC515795  PMID: 196103

Abstract

A nucleoprotein complex that is an intermediate in viral transcription has been isolated from simian virus 40 (SV40)-infected BSC-1 cells after lysing infected nuclei with Sarkosyl. It contain DNA, DNA-dependent RNA polymerase II, and nascent RNA chains. RNA chain elongation continues for several hours in vitro and is dependent on exogenous ribonucleoside triphosphates. The complex sediments in neutral sucrose gradients with a main peak at about 24 to 26S. When the nascent RNA on the complex is treated with RNase A, a fraction of the RNA remains resistant to RNase and is hydrogen bonded to the DNA template. The pulse-labeled RNase-resistant RNA can be chased into RNase-sensitive RNA, indicating that it is located at the 3' terminus of the RNA chain. The rate of RNA displacement from the DNA template is consistent with an average rate of RNA chain elongation of 15 to 30 nucleotides per min. At least 70% of the RNA synthesized in this in vitro system is SV40 specific. Hybridization with the separated strands of SV40 DNA and with fragments of SV40 DNA generated with endonucleases HindII + III indicates that this RNA is complementary to all regions of the "late" SV40 DNA strand. Studies of SV40 RNA synthesis in this partially purified preparation at early and late times after infection should provide a way of locating promoter sites for transcription and identifying the form of SV40 DNA that serves as a template for late transcription.

Full text

PDF
20

Selected References

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

  1. Acheson N. H. Transcription during productive infection with polyoma virus and Simian virus 40. Cell. 1976 May;8(1):1–12. doi: 10.1016/0092-8674(76)90179-3. [DOI] [PubMed] [Google Scholar]
  2. Aloni Y., Shani M., Reuveni Y. RNAs of simian virus 40 in productively infected monkey cells: kinetics of formation and decay in enucleate cells. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2587–2591. doi: 10.1073/pnas.72.7.2587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Attardi B., Attardi G. Expression of the mitochondrial genome in HeLa cells. I. Properties of the discrete RNA components from the mitochondrial fraction. J Mol Biol. 1971 Jan 28;55(2):231–249. doi: 10.1016/0022-2836(71)90194-x. [DOI] [PubMed] [Google Scholar]
  4. Cedar H., Felsenfeld G. Transcription of chromatin in vitro. J Mol Biol. 1973 Jun 25;77(2):237–254. doi: 10.1016/0022-2836(73)90334-3. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Gilboa E., Aviv H. Preferential synthesis of viral late RNA by nuclei isolated from SV40 lytically infected cells. Cell. 1976 Apr;7(4):567–573. doi: 10.1016/0092-8674(76)90207-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. HAYASHI M., SPIEGELMAN S. The selective synthesis of informational RNA in bacteria. Proc Natl Acad Sci U S A. 1961 Oct 15;47:1564–1580. doi: 10.1073/pnas.47.10.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Khoury G., Howley P., Nathans D., Martin M. Posttranscriptional selection of simian virus 40-specific RNA. J Virol. 1975 Feb;15(2):433–437. doi: 10.1128/jvi.15.2.433-437.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laub O., Aloni Y. Transcription of simian virus 40. V. Regulattion of simian virus 40 gene expression. J Virol. 1975 Nov;16(5):1171–1183. doi: 10.1128/jvi.16.5.1171-1183.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lebowitz J., Garon C. G., Chen M. C., Salzman N. P. Chemical modification of simian virus 40 DNA by reaction with a water-soluble carbodiimide. J Virol. 1976 Apr;18(1):205–210. doi: 10.1128/jvi.18.1.205-210.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lebowitz P., Bloodgood R. Transcription of simian virus 40 DNA by Escherichia coli RNA polymerase: synthesis of a DNA-RNA hybrid and discrete RNAs under restrictive transcription conditions. J Mol Biol. 1975 May 15;94(2):183–201. doi: 10.1016/0022-2836(75)90077-7. [DOI] [PubMed] [Google Scholar]
  13. May E., Kopecka H., May P. Mapping the transcription site of the SV40-specific late 16 S mRNA. Nucleic Acids Res. 1975 Oct;2(10):1995–2005. doi: 10.1093/nar/2.10.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meilhac M., Tysper Z., Chambon P. Animal DNA-dependent RNA polymerases. 4. Studies on inhibition by rifamycin derivatives. Eur J Biochem. 1972 Jul 13;28(2):291–300. doi: 10.1111/j.1432-1033.1972.tb01913.x. [DOI] [PubMed] [Google Scholar]
  15. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  16. Richardson J. P. Attachment of nascent RNA molecules to superhelical DNA. J Mol Biol. 1975 Nov 5;98(3):565–579. doi: 10.1016/s0022-2836(75)80087-8. [DOI] [PubMed] [Google Scholar]
  17. Shmookler R. J., Buss J., Green M. H. Properties of the polyoma virus transcription complex obtained from mouse nuclei. Virology. 1974 Jan;57(1):122–127. doi: 10.1016/0042-6822(74)90113-5. [DOI] [PubMed] [Google Scholar]
  18. Weinmann R., Roeder R. G. Role of DNA-dependent RNA polymerase 3 in the transcription of the tRNA and 5S RNA genes. Proc Natl Acad Sci U S A. 1974 May;71(5):1790–1794. doi: 10.1073/pnas.71.5.1790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wilhelm J., Brison O., Kedinger C., Chambon P. Characterization of adenovirus type 2 transcriptional complexes isolated from infected HeLa cell nuclei. J Virol. 1976 Jul;19(1):61–81. doi: 10.1128/jvi.19.1.61-81.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES