title: DNA gyrase with a single catalytic tyrosine can catalyze DNA supercoiling by a nicking-closing mechanism.
abstract: The topological state of DNA is important for replication, recombination and transcription, and is regulated in vivo by DNA topoisomerases. Gyrase introduces negative supercoils into DNA at the expense of ATP hydrolysis. It is the accepted view that gyrase achieves supercoiling by a strand passage mechanism, in which double-stranded DNA is cleaved, and a second double-stranded segment is passed through the gap, converting a positive DNA node into a negative node. We show here that gyrase with only one catalytic tyrosine that cleaves a single strand of its DNA substrate can catalyze DNA supercoiling without strand passage. We propose an alternative mechanism for DNA supercoiling via nicking and closing of DNA that involves trapping, segregation and relaxation of two positive supercoils. In contrast to DNA supercoiling, ATP-dependent relaxation and decatenation of DNA by gyrase lacking the C-terminal domains require both tyrosines and strand passage. Our results point towards mechanistic plasticity of gyrase and might pave the way for finding novel and specific mechanism-based gyrase inhibitors.
authors.affiliation: University of Muenster, Institute for Physical Chemistry, Corrensstrasse 30, D-48149 Muenster, Germany.
authors.forename: Airat
authors.initials: A
authors.lastname: Gubaev
authors.affiliation: University of Muenster, Institute for Physical Chemistry, Corrensstrasse 30, D-48149 Muenster, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Weidlich
authors.affiliation: University of Muenster, Institute for Physical Chemistry, Corrensstrasse 30, D-48149 Muenster, Germany dagmar.klostermeier@uni-muenster.de.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Klostermeier
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology; D002384:Catalysis; D027081:DNA Gyrase / Q000737:chemistry* / Q000378:metabolism*; D004278:DNA, Superhelical / Q000737:chemistry* / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D008968:Molecular Conformation; D013329:Structure-Activity Relationship; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004278:DNA, Superhelical; D014443:Tyrosine; D027081:DNA Gyrase