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Multiple modes of Escherichia coli DNA gyrase activity revealed by force and torque.

Authors
  • M, Nöllmann
  • Michael Stone
  • Z, Bryant
  • J, Gore
  • Nj, Crisona
  • Sc, Hong
  • S, Mitelheiser
  • A, Maxwell
  • C, Bustamante
  • Nr, Cozzarelli
Type
Published Article
Journal
Nature Structural & Molecular Biology
Publisher
Springer Nature
Volume
14
Issue
4
Pages
264–271
Source
UCSC Cancer biomedical-ucsc
License
Unknown

Abstract

E. coli DNA gyrase uses the energy of ATP hydrolysis to introduce essential negative supercoils into the genome, thereby working against the mechanical stresses that accumulate in supercoiled DNA. Using a magnetic-tweezers assay, we demonstrate that small changes in force and torque can switch gyrase among three distinct modes of activity. Under low mechanical stress, gyrase introduces negative supercoils by a mechanism that depends on DNA wrapping. Elevated tension or positive torque suppresses DNA wrapping, revealing a second mode of activity that resembles the activity of topoisomerase IV. This distal T-segment capture mode results in active relaxation of left-handed braids and positive supercoils. A third mode is responsible for the ATP-independent relaxation of negative supercoils. We present a branched kinetic model that quantitatively accounts for all of our single-molecule results and agrees with existing biochemical data.

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