Skip to main content

Eukaryotic Nuclear RNA Polymerases (EC 2.7.7.6)

  • Protocol
Enzymes of Molecular Biology

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 16))

Abstract

DNA-dependent RNA polymerases (ribonucleoside triphosphate: RNA nucleotidyltransferase, EC 2.7.7.6) catalyze the synthesis of RNA from ribonucleoside triphosphates in the presence of a DNA template and divalent cation (1).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Slater, R. J. (1987) In vitro transcription, in Techniques in Molecular Biology, vol. 2 (Walker, J. M. and Gaastra, W., eds.), Groom Helm, London and Sidney, pp. 203–227.

    Google Scholar 

  2. Jacob, S. T. (1973) Mammalian RNA polymerases. Prog. Nucleic Acids Res. Mol. Biol. 13, 93–126.

    Article  CAS  Google Scholar 

  3. Chambon, P. (1975) Eukaryotic nuclear RNA polymerases. Ann. Rev. Biochem. 44, 613–638.

    Article  PubMed  CAS  Google Scholar 

  4. Roeder, R. G. (1976) Eukaryotic nuclear RNA polymerases, in RNA Polymerase (Losick, R. and Chamberlin, M., eds.), Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 285–329.

    Google Scholar 

  5. Chamberlin, M. (1982) Bacterial DNA-dependent RNA polymerases, in The Enzymes, 3rd ed., vol. 15B (Boyer, P., ed.), Academic, New York, pp. 61–108.

    Google Scholar 

  6. Guilfoyle, T. J. (1983) DNA-dependent RNA polymerases of plants and lower eukaryotes, in Enzymes of Nucleic Acid Synthesis and Modification, vol. 2 (Jacob, S. T., ed.), CRC, Cleveland, OH, pp. 1–41.

    Google Scholar 

  7. Beebee, T. J. C. and Butterworth, P. H. W. (1981) Eukaryotic DNA-dependent RNA polymerases: An evaluation of their role in the regulation of gene expression, in Eukaryotic Gene Regulation, vol. 2 (Kolodny, G. M., ed.), CRC, Cleveland, OH, pp. 1–56.

    Google Scholar 

  8. Chandler, D. W. and Gralla, J. (1980) Specific binding and protection of form II SV40 deoxyribonucleic acid by ribonucleic acid polymerase II from wheat germ. Biochemistry 19, 1604–1612.

    Article  PubMed  CAS  Google Scholar 

  9. Seidman, S., Surzycki, S. J., Delorbe, W., and Gussin, G. N. (1979) Interaction between wheat germ RNA polymerase II and adenovirus 2 DNA. Evidence for two types of stable binary complexes. Biochemistry 18, 3363–3371.

    Article  PubMed  CAS  Google Scholar 

  10. Grellet, F., Cooke, R., Teissere, M., Delseny, M., Xech, J., and Penon, P. (1981) Electron microscopic mapping of wheat germ RNA polymerase II binding sites on cloned CaMV DNA. Nucleic Acids Res. 9(16), 3927–3939.

    Article  PubMed  CAS  Google Scholar 

  11. Mildvan, A. S. and Loeb, L. A. (1979). The role of metal ions in the mechanism of DNA and RNA polymerases. CRC. Crit. Rev. Biochem. 6, 219–244.

    Article  PubMed  CAS  Google Scholar 

  12. Chen, Y. M., Lin, C. Y., Chang, H., Guilfoyle, T. J., and Key, J. L. (1975) Isolation and properties of nuclei from control and auxin treated soybean hypocotyl. Plant Physiol. 56, 78–82.

    Article  PubMed  CAS  Google Scholar 

  13. Lin, C. Y., Guilfoyle, T. J., Chen, Y. M., Nagao, R. T., and Key, J. L. (1975) Isolation of nucleoli and localisation of ribonucleic acid polymerase I from soybean hypocotyl. Plant Physiol. 56, 850–852.

    Article  PubMed  CAS  Google Scholar 

  14. Guilfoyle, T. J. and Key, J. L. (1977) Purification and characterization of soybean DNA-dependent RNA polymerases and the modulation of their activities during development, in Nucleic Acids and Protein Synthesis in Plants (Bogorad, L. and Weil, J., eds.), Plenum, New York, pp. 37–63.

    Google Scholar 

  15. Guilfoyle, T. J. (1980) Transcription of the cauliflower mosaic virus genome in isolated nuclei from turnip leaves. Virology 107, 71–80.

    Article  PubMed  CAS  Google Scholar 

  16. Matsui, T., Segall, J., Weil, A., and Roeder, R. G. (1980) Multiple factors for accurate initiation of transcription by purified RNA polymerase II. J. Biol. Chem. 255(24), 11,992–11,996.

    PubMed  CAS  Google Scholar 

  17. Samuels, M., Fire, A., and Sharp, P. A. (1982) Separation and characterization of factors mediating accurate transcription by RNA polymerase II. J. Biol. Chem. 257, 14,419–14,427.

    PubMed  CAS  Google Scholar 

  18. Dignam, J. D., Martin, P. L., Shastry, B. S., and Roeder, R. G. (1983) Eukaryotic gene transcription with purified components. Methods Enzymol. 104, 582–598.

    Article  Google Scholar 

  19. Sawadogo, M. and Roeder, R. G. (1985) Factors involved in specific transcription by human RNA polymerase II: Analysis by a rapid and quantitative in vitro assay. Proc. Natl. Acad. Sci. USA 82, 4394–4398.

    Article  PubMed  CAS  Google Scholar 

  20. Sawadogo, M. and Roeder, R. G. (1985) Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell 43, 165–175.

    Article  PubMed  CAS  Google Scholar 

  21. Nakajima, N., Horikoshi, M., and Roeder, R. G. (1988) Factors involved in specific transcription by mammalian RNA polymerase II. Purification, genetic specificity and TATA box-promoter interaction of TFIID. Cell Biol. 8, 4028–4040.

    CAS  Google Scholar 

  22. Davison, B. L., Egly, J., Mulvihill, E. R., and Chambon, P. (1983) Formation of stable preinitiation complexes between class B transcription factors and promoter sequences. Nature 301, 680–686.

    Article  PubMed  CAS  Google Scholar 

  23. Fire, A., Samuels, M., and Sharp, P. A. (1984) Interactions between RNA polymerase II factors and template leading to accurate transcription. J. Biol. Chem. 259, 2509–2516.

    PubMed  CAS  Google Scholar 

  24. Grosschedl, R. and Birnstiel, M. L. (1980) Spacer DNA sequences upstream of the T-A-T-A-A-A-T-A sequence are essential for promotion of H2A histone gene transcription in vivo. Proc. Natl. Acad. Sci. USA 77, 7102–7106.

    Article  PubMed  CAS  Google Scholar 

  25. Dynan, W. S. and Tjian, R (1985) Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins. Nature 316, 774–778.

    Article  PubMed  CAS  Google Scholar 

  26. Zheng, X-M., Moncollin, V., Egly, J. M., and Chambon, P. (1987) A general transcription factor forms a stable complex with RNA polymerase II. Cell 50, 361–368.

    Article  PubMed  CAS  Google Scholar 

  27. Reinberg, D. and Roeder, R. G. (1987) Factors involved in specific transcription by mammalian RNA polymerase II: Purification and functional analysis of factors IIB and IIE. J. Biol. Chem. 262, 3310–3321.

    PubMed  CAS  Google Scholar 

  28. Buratowski, S., Hahn, S., Sharp, P. A., and Guarente, L. (1988) Function of a yeast TATA element binding protein a mammalian transcription system. Nature 334, 37–42.

    Article  PubMed  CAS  Google Scholar 

  29. Ptashne, M. (1986) Gene regulation by proteins acting nearby and at a distance. Nature 225, 683–689.

    Google Scholar 

  30. Jones, K. A., Yamamoto, K. R., and Tjian, R. (1985) Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro. Cell 42, 559–572.

    Article  PubMed  CAS  Google Scholar 

  31. Jones, K. A., Kadonaga, J. T., Rosenfield, P. J., Kelly, T. J., and Tjian, R. (1987) A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. Cell 48, 79–89.

    Article  PubMed  CAS  Google Scholar 

  32. Kadonaga, J. T., Jones, K. A., and Tjian, R. (1986) Promoter-specific activation of RNA polymerase II transcription by Sp1. TIBS 11, 20–23.

    CAS  Google Scholar 

  33. Sawadogo, M. and Roeder, R. G. (1985) Interaction of a gene specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell 43, 165–169.

    Article  PubMed  CAS  Google Scholar 

  34. Carthew, R W., Chodosh, L. A., and Sharp, P. A (1985) An RNA polymerase II transcription factor binds to an upstream element in the adenovirus major late promoter. Cell 43, 439–444.

    Article  PubMed  CAS  Google Scholar 

  35. Chodosh, L. A., Carthew, R. W., Morgan, J. G., Crabtree, G. R., and Sharp, P. A. (1987) The adenovirus major late transcription factor activates the rat γ-fibrinogen promoter. Science 238, 684–691.

    Article  PubMed  CAS  Google Scholar 

  36. Pelham, H. R. B. (1985) Activation of heat shock genes in eukaryotes. Trends Genet. 1, 31–40.

    Article  CAS  Google Scholar 

  37. Sorger, P. K., Lewis, M. J., and Pelham, H. R. B. (1987) Heat shock factor is regulated differently in yeast and HeLa cells. Nature 329, 81–85.

    Article  PubMed  CAS  Google Scholar 

  38. Weil, P. A., Luse, D. S., Segall, J., and Roeder, R. G. (1979) Selective and accurate initiation of transcription at the Ad2 major late promoter in a soluble system dependent on purified RNA polymerase II and DNA. Cell 18, 469–484.

    Article  PubMed  CAS  Google Scholar 

  39. Manley, J. L., Fire, A., Cano, A., Sharp, P. A., and Gefter, M. L. (1980) DNA dependent transcription of adenovirus genes in a soluble whole cell extract. Proc. Natl. Acad. Sci. USA 77, 3855–3859.

    Article  PubMed  CAS  Google Scholar 

  40. Dignam, J. D., Lebovitz, R. M., and Roeder, R. G. (1983) Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acid. Res. 11, 1475–1489.

    Article  PubMed  CAS  Google Scholar 

  41. Zerivitz, K. and Akusjarvi, G. (1989) An improved nuclear extract preparation method. Gene Anal. Techn. 6, 101–109.

    Article  CAS  Google Scholar 

  42. Wu, G-J. (1978) Adenovirus DNA-directed transcription of 5.5S rRNA in vitro. Proc. Natl. Acad. Sci. USA 75, 2175–2179.

    Article  PubMed  CAS  Google Scholar 

  43. Heintz, N. and Roeder, R. (1982) Transcription of eukaryotic genes in soluble cell-free systems., in Genetic Engineering Principals and Methods, vol. 4 (Setlow, J. and Hollaener, A., eds.), Plenum, New York and London.

    Google Scholar 

  44. Manley, J. L. (1982) Transcription of mammalian genes in vitro, in Genetic Engineering, vol. 4 (Setlow, J. K. and Hollaender, A., eds.), Plenum, New York, pp. 37–56.

    Google Scholar 

  45. McKnight, S. L. and Kisbury, R. (1982) Transcription control signals of a eukaryotic protein coding gene. Science 217, 316–324.

    Article  PubMed  CAS  Google Scholar 

  46. Saiki, R. K. (1989) Basic methodology, in PCR Technology: Principles and Applications for DNA Amplification (Erlich, H. A., ed.), Macmillan, Basingstoke, UK, pp. 7–38.

    Google Scholar 

  47. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  48. Vogt, V. M. (1973) Purification and further properties of single-stranded specific nucleases from Aspergillus Oryzae. Eur. J. Biochem. 33, 192–198.

    Article  PubMed  CAS  Google Scholar 

  49. Berk, A. J. and Sharp, P. A. (1977) Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease digested hybrids. Cell 12, 721–728.

    Article  PubMed  CAS  Google Scholar 

  50. Weaver, R. F. and Weissmann, C. (1979) Mapping by a modification of the Berk-Sharp procedure. 5′ termini of 15s β-globin mRNA precursor and mature 10s β-globin mRNA have identical map coordinates. Nucleic Acids Res. 7, 1175–1180.

    Article  PubMed  CAS  Google Scholar 

  51. Casey, J. and Davidson, N. (1977) Rates of formation and thermal stabilities of RNA:DNA and DNA:DNA duplexes at high concentrations of formamide. Nucleic Acids Res. 4, 1539–1545.

    Article  PubMed  CAS  Google Scholar 

  52. Favalaro, J., Treisman, R., and Kamen, R. (1980) Transcription maps of polyoma virus specific RNA: Analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 65, 718–720.

    Article  Google Scholar 

  53. Manley, J. L., Fire, A., Samules, M., and Sharp, P. A. (1983) In vitro transcription: whole-cell extract. Methods Enzymol. 101, 568–582.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1993 Humana Press Inc.

About this protocol

Cite this protocol

Cook, D.A., Slater, R.J. (1993). Eukaryotic Nuclear RNA Polymerases (EC 2.7.7.6). In: Burrell, M.M. (eds) Enzymes of Molecular Biology. Methods in Molecular Biology™, vol 16. Humana Press. https://doi.org/10.1385/0-89603-234-5:59

Download citation

  • DOI: https://doi.org/10.1385/0-89603-234-5:59

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-322-1

  • Online ISBN: 978-1-59259-503-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics