Tn7 transposition: target DNA recognition is mediated by multiple Tn7-encoded proteins in a purified in vitro system.

We have reconstituted the transposition of the bacterial transposon Tn7 into its specific insertion site attTn7 with four purified Tn7-encoded proteins, TnsA+TnsB+TnsC+TnsD, and ATP. TnsA+TnsB+TnsC form a "core" recombination machine that recognizes the transposon ends and executes DNA breakage and joining; TnsD specifically recognizes attTn7. TnsA+TnsB+TnsC are specifically targeted to attTn7 through the TnsD-dependent interaction of TnsC, a nonspecific DNA-binding protein, with attTn7. Recombination appears to be activated by the assembly of a nucleoprotein complex containing the DNA substrates and Tns proteins. We suggest that TnsC plays a central role in communication between the transposon and the target DNA, particularly in directing insertion away from DNAs already containing a copy of Tn7.