title: Reverse gyrase binding to DNA alters the double helix structure and produces single-strand cleavage in the absence of ATP.
abstract: Stoichiometric amounts of pure reverse gyrase, a type I topoisomerase from the archaebacterium Sulfolobus acidocaldarius were incubated at 75 degrees C with circular DNA containing a single-chain scission. After covalent closure by a thermophilic ligase and removal of bound protein molecules, negatively supercoiled DNA was produced. This finding, obtained in the absence of ATP, contrasts with the ATP-dependent positive supercoiling catalyzed by reverse gyrase and is interpreted as the result of enzyme binding to DNA at high temperature. Another consequence of reverse gyrase stoichiometric binding to DNA is the formation of a cleavable complex which results in the production of single-strand breaks in the presence of detergent. Like eubacterial type I topoisomerase (protein omega), reverse gyrase is tightly attached to the 5' termini of the cleaved DNA. In the light of these results, a comparison is tentatively made between reverse gyrase and the eubacterial type I (omega) and type II (gyrase) topoisomerases.
authors.affiliation: Laboratoire d'Enzymologie des Acides Nucléiques, URA 3 CNRS, Université Pierre et Marie Curie, Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Jaxel
authors.forename: M
authors.initials: M
authors.lastname: Nadal
authors.forename: G
authors.initials: G
authors.lastname: Mirambeau
authors.forename: P
authors.initials: P
authors.lastname: Forterre
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: M
authors.initials: M
authors.lastname: Duguet
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004278:DNA, Superhelical; D004591:Electrophoresis, Polyacrylamide Gel; D009690:Nucleic Acid Conformation; D013696:Temperature
chemical_list: D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D000255:Adenosine Triphosphate; D004247:DNA; D004264:DNA Topoisomerases, Type I; D011088:DNA Ligases