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Nucleic Acids Research, 1989, Vol. 17, No. 15 6043-6054
© 1989

MOLECULAR BIOLOGY

The recombinational enhancer for DNA inversion functions independent of its orientation as a consequence of dyad symmetry in the Fis-DNA complex

Ronald Kanaar*, Jochem P. van Hal and Pieter van de Putte

Laboratory of Molecular Genetics, Department of Biochemistry, Leiden University 2333 AL Leiden, The Netherlands

Received May 8, 1989. Revised July 11, 1989. Accepted July 11, 1989.

The Escherichia coli Fis protein binds to specific DNA sequences whose base composition varies enormously. One known function of Fis is to stimulate site-specific DNA recombination. We used the Gin-mediated DNA inversion system of bacteriophage Mu to analyze Fis-DNA interaction. Efficient inversion requires an enhancer which consists of two Fis binding sites at a fixed distance from each other. Using mutant enhancers in which one of the Fis binding sites is replaced we show that Fis binds symmetrically to the DNA and we locate the center of symmetry. Furthermore, we show that one of the Fis binding sites can be replaced by a Fis binding site that normally functions in a process other than site-specific recombination.

*Present address: Department of Molecular Biology. University of California, Berkeley, CA 94720. USA


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