Direct observation of DNA rotation during transcription by Escherichia coli RNA polymerase

Nature. 2001 Jan 4;409(6816):113-5. doi: 10.1038/35051126.

Abstract

Helical filaments driven by linear molecular motors are anticipated to rotate around their axis, but rotation consistent with the helical pitch has not been observed. 14S dynein and non-claret disjunctional protein (ncd) rotated a microtubule more efficiently than expected for its helical pitch, and myosin rotated an actin filament only poorly. For DNA-based motors such as RNA polymerase, transcription-induced supercoiling of DNA supports the general picture of tracking along the DNA helix. Here we report direct and real-time optical microscopy measurements of rotation rate that are consistent with high-fidelity tracking. Single RNA polymerase molecules attached to a glass surface rotated DNA for >100 revolutions around the right-handed screw axis of the double helix with a rotary torque of >5 pN nm. This real-time observation of rotation opens the possibility of resolving individual transcription steps.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry
  • DNA / metabolism*
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / metabolism*
  • Escherichia coli / enzymology
  • Fluorescent Dyes
  • Magnetics
  • Microscopy
  • Microspheres
  • Molecular Motor Proteins
  • Nucleic Acid Conformation
  • RNA / metabolism
  • Rotation
  • Transcription, Genetic*

Substances

  • Fluorescent Dyes
  • Molecular Motor Proteins
  • RNA
  • DNA
  • DNA-Directed RNA Polymerases