Letter | Published:

Transcription by single molecules of RNA polymerase observed by light microscopy

Naturevolume 352pages444448 (1991) | Download Citation

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Abstract

THE kinetics of transcription by Escherichia coli RNA polymerase relate directly to the regulation of transcription and to the properties of processive enzymes in general1, but analysis of RNA polymerase movement along the DNA template has so far been limited to the study of populations of enzyme molecules. The ability to view nanometre-sized particles with the light microscope2,3 suggested a method of monitoring transcription by individual RNA polymerase molecules. We describe here the behaviour of 40-nm-diameter particles of colloidal gold attached to the ends of DNA molecules being transcribed by RNA polymerase immobilized on a glass surface. The tethered gold particles are released from the surface at times after addition of nucleoside triphosphates that are consistent with the kinetics of transcription by RNA polymerase in solution. Analysis of the brownian motion of the gold particles enabled us to measure the movement along the template DNA of individual polymerase molecules.

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References

  1. 1

    Yager, T. D. & von Hippel, P. H. in Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology (eds F. C. Neidhardt et al.) 1241–1275 (American Society for Microbiology, Washington, DC, 1989).

  2. 2

    Geerts, H. et al. Biophys. J. 52, 775–782 (1987).

  3. 3

    Sheetz, M. P., Turney, S., Qian, H. & Elson, E. L. Nature 340, 284–288 (1989).

  4. 4

    Lee, D. N., Phung, L., Stewart, J. & Landick, R. J. biol. Chem. 265, 15145–15153 (1990).

  5. 5

    Levin, J. R., Krummel, B. & Chamberlin, M. J. J. molec. Biol. 196, 85–100 (1987).

  6. 6

    Landick, R. & Yanofsky, C. J. molec. Biol. 196, 363–377 (1987).

  7. 7

    Metzger, W., Schicktor, P. & Heumann, H. EMBO J. 8, 2745–2754 (1989).

  8. 8

    Krummel, B. & Chamberlin, M. J. Biochemistry 28, 7829–7842 (1989).

  9. 9

    Saenger, W. Principles of Nucleic Acid Structure (Springer, New York, 1984).

  10. 10

    Higuchi, R., Krumel, B. & Saiki, R. K. Nucleic Acids Res. 16, 7351–7367 (1988).

  11. 11

    Gelles, J., Schnapp, B. J. & Sheetz, M. P. Nature 331, 450–453 (1988).

  12. 12

    Allen, R. D. & Allen, N. S. J. Microsc. 129, 3–17 (1983).

  13. 13

    Press, W. H., Flannery, B. P., Teukolsky, S. A. & Vetterling, W. T. Numerical Recipes in C (Cambridge University Press, Cambridge, 1988).

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Author notes

  1. Robert Landick: To whom correspondence should be addressed.

Affiliations

  1. Department of Cell Biology, Washington University, St Louis, Missouri, 63130, USA

    • Dorothy A. Schafer
  2. Department of Biology, Washington University, St Louis, Missouri, 63130, USA

    • Robert Landick
  3. Graduate Department of Biochemistry and The Center for Complex Systems, Brandeis University, Waltham, Massachusetts, 02254, USA

    • Jeff Gelles
  4. Department of Cell Biology, Duke University Medical Center, Durham, North Carolina, 27710, USA

    • Michael P. Sheetz

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https://doi.org/10.1038/352444a0

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