title: Eukaryotic topoisomerases recognize nucleic acid topology by preferentially interacting with DNA crossovers.
abstract: Eukaryotic topoisomerases recognize DNA topology and preferentially react with positively or negatively supercoiled molecules over relaxed substrates. To elucidate the mechanism of this recognition, we examined the interaction of topoisomerases with DNA by electron microscopy. Under all conditions employed, approximately 90% of the bound type I or II enzyme was observed at points of helix--helix juxtaposition on negatively supercoiled plasmids which contained as few as four crossovers. Recognition was independent of torsional stress, as enzyme molecules were also found at crossovers on linear DNA. Since juxtaposed helices are more prevalent in supercoiled compared with relaxed nucleic acids, we propose that eukaryotic topoisomerases I and II recognize underwound or overwound substrates by interacting preferentially with DNA crossovers. This may represent a general mechanism for the recognition of DNA topology by proteins.
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232-0146.
authors.forename: E L
authors.initials: EL
authors.lastname: Zechiedrich
authors.forename: N
authors.initials: N
authors.lastname: Osheroff
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology*; D004247:DNA / Q000378:metabolism / Q000648:ultrastructure*; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004331:Drosophila melanogaster; D009690:Nucleic Acid Conformation
chemical_list: D004247:DNA; D004264:DNA Topoisomerases, Type I; D004250:DNA Topoisomerases, Type II