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Nucleic Acids Research, 1981, Vol. 9, No. 16 4201-4210
© 1981

MOLECULAR BIOLOGY

Genetic recombination: recA protein promotes homologous pairing between duplex DNA molecules without strand unwinding

Era Cassuto, Stephen C. West, Jill Podell and Paul Howard-Flanders

Department of Molecular Biophysics and Biochemistry, and Department of Therapeutic Radiology Yale University, Box 6666, New Haven, CT 06511, USA

Received March 30, 1981. The recA protein of Escherichia coli promotes pairing in vitro between covalent circular duplex DNA and homologous circular duplex DNA containing a single stranded region. We have used a filter binding assay to investigate the frequency of homologous pairing between gapped and intact duplex DNA when unwinding of the free 3' and 5' ends of the gapped molecules was blocked. In order to obtain DNA without free ends, the gapped DNA was treated with trimethylpsoralen and 360 nm light so as to introduce about 6 crosslinks per DNA molecule and the double stranded regions on either side of the gaps were then digested up to the first crosslinks with exonuclease III and {gamma} exonuclease. This treatment did not diminish the frequency of homologous pairing, an observation which is difficult to reconcile with models for recombination requiring strand unwinding before pairing.


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