Genetic analysis of the terminal 8-bp inverted repeats of transposon Tn7

Mutations in the terminal 8-bp (5'-T1G2T3G4G5G6C7G8-3') of the inverted repeats of the bacterial transposon, Tn7, were analysed by measuring Tn7 transposition to the attachment site, attTn7. The mutation, C2, present at either end of Tn7 reduces transposition only threefold, but in the double mutant, with C2 at both ends of Tn7, no transposition is detected. C6 mutations have no effect on transposition frequency. Replacement with 5'-A3C4G5C6G7C8-3' at the right end of Tn7 apparently abolishes transposition; yet in the double mutant, where the inverted repeats are restored by substituting this sequence at both ends of Tn7, transposition is partially rescued. This suggests that the mechanism of Tn7 transposition requires communication between the two ends. Tn7 transposition has always been seen to generate a 5-bp target duplication. This is presumed to result from a staggered cut, plus repair synthesis during transposition. We found that two of our right-end mutants, C2 and C6, sometimes yielded a 6-bp target duplication. This observation implies that cleavage of the target site might also involve interaction with the donor ends which, when mutant, relax the specificity for target-site cleavage.