title: Cyclic changes in the affinity of protein-DNA interactions drive the progression and regulate the outcome of the Tn10 transposition reaction.
abstract: The Tn10 transpososome is a DNA processing machine in which two transposon ends, a transposase dimer and the host protein integration host factor (IHF), are united in an asymmetrical complex. The transitions that occur during one transposition cycle are not limited to chemical cleavage events at the transposon ends, but also involve a reorganization of the protein and DNA components. Here, we demonstrate multiple pathways for Tn10 transposition. We show that one series of events is favored over all others and involves cyclic changes in the affinity of IHF for its binding site. During transpososome assembly, IHF is bound with high affinity. However, the affinity for IHF drops dramatically after cleavage of the first transposon end, leading to IHF ejection and unfolding of the complex. The ejection of IHF promotes cleavage of the second end, which is followed by restoration of the high affinity state which in turn regulates target interactions.
authors.affiliation: Department of Biochemistry, University of Oxford South Parks Road, Oxford OX1 3QU, UK.
authors.forename: Danxu
authors.initials: D
authors.lastname: Liu
authors.forename: Paul
authors.initials: P
authors.lastname: Crellin
authors.forename: Ronald
authors.initials: R
authors.lastname: Chalmers
mesh_terms: D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004251:DNA Transposable Elements; D034982:Integration Host Factors / Q000378:metabolism*; D007700:Kinetics; D008957:Models, Genetic; D016254:Mutagenesis, Insertional; D009690:Nucleic Acid Conformation; D010507:Periodicity; D019895:Transposases / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D034982:Integration Host Factors; C473501:Tn10 transposase; D004247:DNA; D019895:Transposases