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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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).
Geerts, H. et al. Biophys. J. 52, 775–782 (1987).
Sheetz, M. P., Turney, S., Qian, H. & Elson, E. L. Nature 340, 284–288 (1989).
Lee, D. N., Phung, L., Stewart, J. & Landick, R. J. biol. Chem. 265, 15145–15153 (1990).
Levin, J. R., Krummel, B. & Chamberlin, M. J. J. molec. Biol. 196, 85–100 (1987).
Landick, R. & Yanofsky, C. J. molec. Biol. 196, 363–377 (1987).
Metzger, W., Schicktor, P. & Heumann, H. EMBO J. 8, 2745–2754 (1989).
Krummel, B. & Chamberlin, M. J. Biochemistry 28, 7829–7842 (1989).
Saenger, W. Principles of Nucleic Acid Structure (Springer, New York, 1984).
Higuchi, R., Krumel, B. & Saiki, R. K. Nucleic Acids Res. 16, 7351–7367 (1988).
Gelles, J., Schnapp, B. J. & Sheetz, M. P. Nature 331, 450–453 (1988).
Allen, R. D. & Allen, N. S. J. Microsc. 129, 3–17 (1983).
Press, W. H., Flannery, B. P., Teukolsky, S. A. & Vetterling, W. T. Numerical Recipes in C (Cambridge University Press, Cambridge, 1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schafer, D., Gelles, J., Sheetz, M. et al. Transcription by single molecules of RNA polymerase observed by light microscopy. Nature 352, 444–448 (1991). https://doi.org/10.1038/352444a0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/352444a0
- Springer Nature Limited
This article is cited by
-
Single-cell measurement of plasmid copy number and promoter activity
Nature Communications (2021)
-
Continuous biomarker monitoring by particle mobility sensing with single molecule resolution
Nature Communications (2018)
-
Helicase promotes replication re-initiation from an RNA transcript
Nature Communications (2018)
-
Real-time assembly of ribonucleoprotein complexes on nascent RNA transcripts
Nature Communications (2018)
-
Diverse architectural properties of Sso10a proteins: Evidence for a role in chromatin compaction and organization
Scientific Reports (2016)