Multiple modes of Escherichia coli DNA gyrase activity revealed by force and torque.
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.
Nöllmann
Marcelo
M
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
Stone
Michael D
MD
Bryant
Zev
Z
Gore
Jeff
J
Crisona
Nancy J
NJ
Hong
Seok-Cheol
SC
Mitelheiser
Sylvain
S
Maxwell
Anthony
A
Bustamante
Carlos
C
Cozzarelli
Nicholas R
NR
D000255:Adenosine Triphosphate / Q000494:pharmacology; D027081:DNA Gyrase / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry; D004278:DNA, Superhelical / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology*; D008280:Magnetics; D008954:Models, Biological; D009690:Nucleic Acid Conformation / Q000187:drug effects; D021122:Protein Subunits / Q000378:metabolism; D019415:Torque
D004269:DNA, Bacterial; D004278:DNA, Superhelical; D021122:Protein Subunits; D000255:Adenosine Triphosphate; D027081:DNA Gyrase