Transposase promotes double strand breaks and single strand joints at Tn10 termini in vivo.
We present evidence that Tn10 transposase promotes double strand breaks and single strand joints at Tn10 termini in vivo. Plasmids containing a shortened Tn10 element and a transposase overproducer fusion give rise, upon transposase induction, to new DNA species. The most prominent class is a circularized transposon molecule whose structure suggests that it arises from double strand breakage at the two transposon ends followed by covalent joining between the 3' and 5' ends of one of the two strands. We have used formation of the circularized transposon as a physical assay for the interaction between transposase and different mutant and wild-type termini. These experiments show that transposase protein interacts preferentially with the genetically most active termini in a way that suppresses productive interaction with weaker termini present on the same substrate molecule.
D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010957:Plasmids; D019895:Transposases
D004251:DNA Transposable Elements; D009713:Nucleotidyltransferases; D019895:Transposases; D004262:DNA Restriction Enzymes