Research
A model for the molecular organisation of the IS911 transpososome
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* Corresponding author: Mick Chandler mike@ibcg.biotoul.fr
- † Equal contributors
1 Université de Toulouse; UPS; Laboratoire de Microbiologie et Génétique Moléculaires, F-31000 Toulouse, France
2 Centre National de la Recherche Scientifique, LMGM, F-31000 Toulouse, France
3 Université de Toulouse; UPS; Institut de Pharmacologie et de Biologie Structurale, F-31000 Toulouse, France
4 Centre National de la Recherche Scientifique, IPBS, F-31000 Toulouse, France
Mobile DNA 2010, 1:16 doi:10.1186/1759-8753-1-16
Published: 16 June 2010Abstract
Tight regulation of transposition activity is essential to limit damage transposons may cause by generating potentially lethal DNA rearrangements. Assembly of a bona fide protein-DNA complex, the transpososome, within which transposition is catalysed, is a crucial checkpoint in this regulation. In the case of IS911, a member of the large IS3 bacterial insertion sequence family, the transpososome (synaptic complex A; SCA) is composed of the right and left inverted repeated DNA sequences (IRR and IRL) bridged by the transposase, OrfAB (the IS911-encoded enzyme that catalyses transposition). To characterise further this important protein-DNA complex in vitro, we used different tagged and/or truncated transposase forms and analysed their interaction with IS911 ends using gel electrophoresis. Our results allow us to propose a model in which SCA is assembled with a dimeric form of the transposase. Furthermore, we present atomic force microscopy results showing that the terminal inverted repeat sequences are probably assembled in a parallel configuration within the SCA. These results represent the first step in the structural description of the IS911 transpososome, and are discussed in comparison with the very few other transpososome examples described in the literature.