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Nucleic Acids Research, 1994, Vol. 22, No. 25 5613-5620
© 1994

Articles

Interactions of the site-specific recombinases XerC and XerD with the recombination site dif

Garry W. BIakely and David J. Sherratt*

Microbiology Unit, Department of Biochemistry, University of Oxford South Parks Road, Oxford 0X1 3QU, UK

*To whom correspondence should be addressed

Received September 20, 1994. Revised November 18, 1994. Accepted November 18, 1994.

The Xer site-specific recombination system of Escherichia coll Is involved in the stable Inheritance of circular replicons. Multimeric repllcons, produced by homologous recombination, are converted to monomers by the action of two related recombinases XerC and XerD. Site-specific recombination at a locus, dif, within the chromosomal replication terminus region is thought to convert dlmerlc chromosomes to monomers, which can then be segregated prior to cell division. The recombinases XerC and XerD bind cooperatively to dif, where they catalyse recombination. Chemical modification of specific bases and the phosphate - sugar backbone within dif was used to Investigate the requirements for binding of the recombinases. Site-directed mutagenesls was then used to alter bases Implicated in recomblnase binding. Characterization of these mutants by In vitro recom-binase binding and In vivo recombination, has demonstrated that the cooperative interactions between XerC and XerD can partially overcome DNA alterations that should interfere with specific recombinase - dif Interactions.


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