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. 1978 Jan;5(1):205–219. doi: 10.1093/nar/5.1.205

Initiation of transcription in nuclei isolated from adenovirus infected cells.

B Vennström, U Pettersson, L Philipson
PMCID: PMC341972  PMID: 643609

Abstract

Initiation of adenovirus transcription was analyzed by incubation of isolated nuclei from virus infected cells in the presence of beta-32P GTP or beta-32P ATP. Nucleotide analysis of RNA from nuclei incubated with beta-32P GTP shows that the label is incorporated exclusively into pppGp and ppGp. Under similar incubation conditions, the label from beta-32P ATP was incorporated primarily into the 5' phosphate of 5', 3' mononucleoside diphosphates, but label was also detected in pppAp, pppGp and in the 3' nucleoside monophosphates. Analysis of RNA, synthesized in the presence of different concentrations of alpha-amanitin, shows that only RNA polymerase III initiates virus specific transcription in isolated nuclei. The virus specific transcripts containing pppAp and pppGp in their 5' termini were identified as the 5.5S and 5.2S viral RNA species by hybridization and finger printing.

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Selected References

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

  1. Forget B. G., Weissman S. M. Nucleotide sequence of KB cell 5S RNA. Science. 1967 Dec 29;158(3809):1695–1699. doi: 10.1126/science.158.3809.1695. [DOI] [PubMed] [Google Scholar]
  2. Gilboa E., Soreq H., Aviv H. Initiation of RNA synthesis in isolated nuclei. Eur J Biochem. 1977 Jul 15;77(2):393–400. doi: 10.1111/j.1432-1033.1977.tb11679.x. [DOI] [PubMed] [Google Scholar]
  3. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  4. Marzluff W. F., Jr, Murphy E. C., Jr, Huang R. C. Transcription of ribonucleic acid in isolated mouse myeloma nuclei. Biochemistry. 1973 Aug 28;12(18):3440–3446. doi: 10.1021/bi00742a013. [DOI] [PubMed] [Google Scholar]
  5. Marzluff W. F., Jr, Murphy E. C., Jr, Huang R. C. Transcription of the genes for 5S ribosomal RNA and transfer RNA in isolated mouse myeloma cell nuclei. Biochemistry. 1974 Aug 27;13(18):3689–3696. doi: 10.1021/bi00715a011. [DOI] [PubMed] [Google Scholar]
  6. Mathews M. B. Genes for VA-RNA in adenovirus 2. Cell. 1975 Oct;6(2):223–229. doi: 10.1016/0092-8674(75)90013-6. [DOI] [PubMed] [Google Scholar]
  7. Ohe K., Weissman S. M. The nucleotide sequence of a low molecular weight ribonucleic acid from cells infected with adenovirus 2. J Biol Chem. 1971 Nov 25;246(22):6991–7009. [PubMed] [Google Scholar]
  8. Price R., Penman S. A distinct RNA polymerase activity, synthesizing 5-5 s, 5 s and 4 s RNA in nuclei from adenovirus 2-infected HeLa cells. J Mol Biol. 1972 Oct 14;70(3):435–450. doi: 10.1016/0022-2836(72)90551-7. [DOI] [PubMed] [Google Scholar]
  9. Price R., Penman S. Transcription of the adenovirus genome by an -amanitine-sensitive ribonucleic acid polymerase in HeLa cells. J Virol. 1972 Apr;9(4):621–626. doi: 10.1128/jvi.9.4.621-626.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Reeder R. H., Roeder R. G. Ribosomal RNA synthesis in isolated nuclei. J Mol Biol. 1972 Jun 28;67(3):433–441. doi: 10.1016/0022-2836(72)90461-5. [DOI] [PubMed] [Google Scholar]
  11. Reichard P., Eliasson R., Söderman G. Initiator RNA in discontinuous polyoma DNA synthesis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4901–4905. doi: 10.1073/pnas.71.12.4901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Söderlund H., Pettersson U., Vennström B., Philipson L., Mathews M. B. A new species of virus-coded low molecular weight RNA from cells infected with adenovirus type 2. Cell. 1976 Apr;7(4):585–593. doi: 10.1016/0092-8674(76)90209-9. [DOI] [PubMed] [Google Scholar]
  14. Teraoka H., Mizuta K., Sato F., Shimoyachi M., Tsukada K. Polynucleotide kinase from rat-liver nuclei. Purification and properties. Eur J Biochem. 1975 Oct 15;58(2):297–302. doi: 10.1111/j.1432-1033.1975.tb02376.x. [DOI] [PubMed] [Google Scholar]
  15. Vennström B., Philipson L. Fidelity of adenovirus RNA transcription in isolated HeLa cell nuclei. J Virol. 1977 May;22(2):290–299. doi: 10.1128/jvi.22.2.290-299.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Volckaert G., Jou W. M., Fiers W. Analysis of 32P-labeled bacteriophage MS2 RNA by a mini-fingerprinting procedure. Anal Biochem. 1976 May 7;72:433–446. doi: 10.1016/0003-2697(76)90551-0. [DOI] [PubMed] [Google Scholar]
  17. Zylber E. A., Penman S. Products of RNA polymerases in HeLa cell nuclei. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2861–2865. doi: 10.1073/pnas.68.11.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]

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