IHF-independent assembly of the Tn10 strand transfer transpososome: implications for inhibition of disintegration.
The frequency of DNA transposition in transposition systems that employ a strand transfer step may be significantly affected by the occurrence of a disintegration reaction, a reaction that reverses the strand transfer event. We have asked whether disintegration occurs in the Tn10 transposition system. We show that disintegration substrates (substrates constituting one half of the strand transfer product) are assembled into a transpososome that mimics the strand transfer intermediate. This strand transfer transpososome (STT) does appear to support an intermolecular disintegration reaction, but only at a very low level. Strikingly, assembly of the STT is not dependent on IHF, a host protein that is required for de novo assembly of all previously characterized Tn10 transpososomes. We suggest that disintegration substrates are able to form both transposon end and target type contacts with transposase because of their enhanced conformational flexibility. This probably allows the conformation of DNA within the complex that prevents the destructive disintegration reaction, and is responsible for relaxing the DNA sequence requirements for STT formation relative to other Tn10 transpososomes.
Stewart
Barry J
BJ
Department of Biochemistry, University of Western Ontario, London, Ontario, Canada N6A 5C1.
Wardle
Simon J
SJ
Haniford
David B
DB
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000378:metabolism
D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D011506:Proteins; D004247:DNA