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Nucleic Acids Research, 2003, Vol. 31, No. 3 1038-1044
© 2003 Oxford University Press

Synapsis and strand exchange in the resolution and DNA inversion reactions catalysed by the ß recombinase

Inés Canosa, Gema López, Fernando Rojo, Martin R. Boocock1 and Juan C. Alonso*

Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus de la Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain and 1 Institute of Biomedical and Life Sciences, University of Glasgow, 56 Dumbarton Road, Glasgow G11 6NU, UK

*To whom correspondence should be addressed. Tel: +34 91585 4546; Fax: +34 91585 4506; Email: jcalonso{at}cnb.uam.es
Present address:
Inés Canosa, Laboratorio Andaluz de Biología, Universidad Pablo de Olavide. Ctra. de Utrera, Km 1, 41013 Sevilla, Spain

In the presence of a sequence-independent chromatin-associated protein, such as Hbsu or HMGB, the ß recombinase catalyses resolution between two directly oriented recombination sites (six sites) and both resolution and DNA inversion between two inversely oriented six sites. Assembly of the synaptic complex requires binding of the ß recombinase to the six sites and the presence of Hbsu. Whether resolution or inversion will take place depends on the relative orientation of the two six sites, the level of DNA supercoiling and the amounts of Hbsu. In this work, the topologies of the products of the resolution and inversion reactions were analysed. The resolution reaction generated mainly singly catenated DNA circles, while DNA inversion gave rise to unknotted circles and small amounts of DNA molecules containing 3- or 5-noded knots. In spite of the distinctive features of the ß system, the topology of synapsis and strand exchange during the resolution reaction is similar to that of Tn3 and {gamma}{delta} resolvases. The ability of the ß recombinase to catalyse both inversion and resolution reactions probably reflects different possible architectures of the synaptic complex, which to a large extent depends on Hbsu.


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