Tn5 transposase loops DNA in the absence of Tn5 transposon end sequences.

Abstract:

:Transposases mediate transposition first by binding specific DNA end sequences that define a transposable element and then by organizing protein and DNA into a highly structured and stable nucleoprotein 'synaptic' complex. Synaptic complex assembly is a central checkpoint in many transposition mechanisms. The Tn5 synaptic complex contains two Tn5 transposase subunits and two Tn5 transposon end sequences, exhibits extensive protein-end sequence DNA contacts and is the node of a DNA loop. Using single-molecule and bulk biochemical approaches, we found that Tn5 transposase assembles a stable nucleoprotein complex in the absence of Tn5 transposon end sequences. Surprisingly, this end sequence-independent complex has structural similarities to the synaptic complex. This complex is the node of a DNA loop; transposase dimerization and DNA specificity mutants affect its assembly; and it likely has the same number of proteins and DNA molecules as the synaptic complex. Furthermore, our results indicate that Tn5 transposase preferentially binds and loops a subset of non-Tn5 end sequences. Assembly of end sequence-independent nucleoprotein complexes likely plays a role in the in vivo downregulation of transposition and the cis-transposition bias of many bacterial transposases.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Adams CD,Schnurr B,Skoko D,Marko JF,Reznikoff WS

doi

10.1111/j.1365-2958.2006.05471.x

subject

Has Abstract

pub_date

2006-12-01 00:00:00

pages

1558-68

issue

6

eissn

0950-382X

issn

1365-2958

pii

MMI5471

journal_volume

62

pub_type

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