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. 1975 Jan;72(1):194–199. doi: 10.1073/pnas.72.1.194

Chromatin as a template for RNA synthesis in vitro.

Y Groner, G Monroy, M Jacquet, J Hurwitz
PMCID: PMC432269  PMID: 1090933

Abstract

RNA transcribed in vitro from myeloblast chromatin by exogenously added RNA polymerase B predominantly consists of short chains that remain in hybrid structure with the template; the remainder of the product is free RNA of heterogeneous size. Addition of polyanions during synthesis caused an increase in the size and amount of free RNA with a concomitant decrease in the proportion of small RNA. The large molecular weight RNA is derived from the short RNA chains, which are synthesized de novo during the reaction in vitro. The effect of polyanions on the size and nature of the product may be related to structural changes induced in the template rather than to an inhibition of nuclease activity.

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Selected References

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

  1. Berkower I., Leis J., Hurwitz J. Isolation and characterization of an endonuclease from Escherichia coli specific for ribonucleic acid in ribonucleic acid-deoxyribonucleic acid hybrid structures. J Biol Chem. 1973 Sep 10;248(17):5914–5921. [PubMed] [Google Scholar]
  2. 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]
  3. Crick F. General model for the chromosomes of higher organisms. Nature. 1971 Nov 5;234(5323):25–27. doi: 10.1038/234025a0. [DOI] [PubMed] [Google Scholar]
  4. Garapin A. C., Leong J., Fanshier L., Levinson W. E., Bishop J. M. Identification of virus-specific RNA in cells infected with Rous sarcoma virus. Biochem Biophys Res Commun. 1971 Mar 5;42(5):919–925. doi: 10.1016/0006-291x(71)90518-3. [DOI] [PubMed] [Google Scholar]
  5. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  6. Hill M., Hillova J. Recovery of the temperature-sensitive mutant of Rous sarcoma virus from chicken cells exposed to DNA extracted from hamster cells transformed by the mutant. Virology. 1972 Jul;49(1):309–313. doi: 10.1016/s0042-6822(72)80034-5. [DOI] [PubMed] [Google Scholar]
  7. Hossenlopp P., Oudet P., Chambon P. Animal DNA-dependent RNA polymerases. Studies on the binding of mammalian RNA polymerases AI and B to Simian virus 40 DNA. Eur J Biochem. 1974 Jan 16;41(2):397–411. doi: 10.1111/j.1432-1033.1974.tb03281.x. [DOI] [PubMed] [Google Scholar]
  8. Howk R. S., Williams D. R., Haberman A. B., Parks W. P., Scolnick E. M. A comparison of the transcription of mouse cell chromatin by homologous RNA polymerase I and II. Cell. 1974 Sep;3(1):15–22. doi: 10.1016/0092-8674(74)90032-4. [DOI] [PubMed] [Google Scholar]
  9. Jacquet M., Groner Y., Monroy G., Hurwitz J. The in vitro synthesis of avian myeloblastosis viral RNA sequences. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3045–3049. doi: 10.1073/pnas.71.8.3045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mandel J. L., Chambon P. Animal DNA-dependent RNA polymerases. Analysis of the RNAs synthesized on Simian virus 40 superhelical DNA by mammalian RNA polymerases AI and B. Eur J Biochem. 1974 Jan 16;41(2):379–395. doi: 10.1111/j.1432-1033.1974.tb03280.x. [DOI] [PubMed] [Google Scholar]
  11. Palmiter R. D. Ovalbumin messenger ribonucleic acid translation. Comparable rates of polypeptide initiation and elongation on ovalbumin and globin messenger ribonucleic acid in a rabbit reticulocyte lysate. J Biol Chem. 1973 Mar 25;248(6):2095–2106. [PubMed] [Google Scholar]
  12. Rabin E. Z., Fraser M. J. Isolation of Neurospora crassa endonuclease specific for single-stranded DNA. Can J Biochem. 1970 Mar;48(3):389–392. doi: 10.1139/o70-063. [DOI] [PubMed] [Google Scholar]
  13. Rhoads R. E., McKnight G. S., Schimke R. T. Quantitative measurement of ovalbumin messenger ribonucleic acid activity. Localization in polysomes, induction by estrogen, and effect of actinomycin D. J Biol Chem. 1973 Mar 25;248(6):2031–2039. [PubMed] [Google Scholar]
  14. Roeder R. G., Rutter W. J. Multiple forms of DNA-dependent RNA polymerase in eukaryotic organisms. Nature. 1969 Oct 18;224(5216):234–237. doi: 10.1038/224234a0. [DOI] [PubMed] [Google Scholar]
  15. Rymo L., Parsons J. T., Coffin J. M., Weissmann C. In vitro synthesis of Rous sarcoma virus-specific RNA is catalyzed by a DNA-dependent RNA polymerase. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2782–2786. doi: 10.1073/pnas.71.7.2782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sigal N., Delius H., Kornberg T., Gefter M. L., Alberts B. A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3537–3541. doi: 10.1073/pnas.69.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stein H., Hausen P. Enzyme from calf thymus degrading the RNA moiety of DNA-RNA Hybrids: effect on DNA-dependent RNA polymerase. Science. 1969 Oct 17;166(3903):393–395. doi: 10.1126/science.166.3903.393. [DOI] [PubMed] [Google Scholar]
  18. WEIL R., VINOGRAD J. THE CYCLIC HELIX AND CYCLIC COIL FORMS OF POLYOMA VIRAL DNA. Proc Natl Acad Sci U S A. 1963 Oct;50:730–738. doi: 10.1073/pnas.50.4.730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yarbrough L. R., Hurwitz J. The reversible denaturation of deoxyribonucleic acid-dependent ribonucleic acid polymerase of Escherichia coli. J Biol Chem. 1974 Sep 10;249(17):5394–5399. [PubMed] [Google Scholar]

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