DNA sequences at the ends of transposon Tn5 required for transposition

Abstract

Transposons are a class of genetic elements that can move from one site in a cell's genome to another independently of the cell's general recombination system. Little is known about the mechanism of transposition of compound transposons such as Tn5, but it is thought that a transposon-encoded protein (a transposase) must recognize the outer ends of the element and, together with host factors, catalyse the transfer of the internal DNA into a new site in a manner that may involve replication. It has previously been shown that the synthesis of an IS50R-encoded protein (protein 1) is an essential requirement for Tn5 transposition^1–5. Here we demonstrate that a structure containing only the outer 186 base pairs (bp) of both inverted repeats is capable of being efficiently complemented to transpose in Escherichia coli, provided IS50R is located close by on the same replicon. In addition, Bal31-generated deletions indicate that 16–18 bp of the outer end of IS50L are required for transposition. This 16–18-bp sequence contains the 8-9-bp small inverted repeat present at each end of IS50 plus a 9–bp sequence which is homologous to an interrelated sequence present in four copies in the chromosomal origin of replication in a variety of Gram-negative bacteria. This sequence organization suggests that the ends of Tn5 may function to provide a recognition site for the Tn5 transposase adjacent to a sequence recognized by the host replication system.