Transcription by single molecules of RNA polymerase observed by light microscopy

Dorothy A. Schafer, Jeff Gelles, Michael Sheetz, Robert Landick

Research output: Contribution to journalArticlepeer-review

208 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)444-448
Number of pages5
JournalNature
Volume352
Issue number6334
DOIs
StatePublished - Jan 1 1991
Externally publishedYes

ASJC Scopus subject areas

  • General

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