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J. Biol. Chem., Vol. 265, Issue 17, 10156-10163, 06, 1990
EC Conley and SC West
Homologous pairing between gapped circular and partially homologous form I
DNA, catalyzed by Escherichia coli RecA protein, leads to the formation of
nascent synaptic joints between regions of duplex DNA. These duplex-duplex
interactions result in underwinding of the form I DNA, as detected by a
topoisomerase assay. Underwound DNA species have been studied with regard
to their formation, stability, and topological requirements. The synaptic
joints are short-lived and of low frequency compared with those formed
between single-stranded and duplex DNA. Measurement of the degree of
underwinding indicates joints 300-400 base pairs in length, in which the
two DNA molecules are presumed to be interwound within the
RecA-nucleoprotein filament. Underwound DNA was not detected in reactions
between gapped DNA and partially homologous nicked circular or relaxed
covalently closed DNA. We have also investigated the requirements for the
initiation of strand exchange. Previous results have shown that strand
exchange requires a homologous 3'-terminus complementary to the gapped
region. We now show that the minimum length of overlap required for
efficient initiation of strand exchange is one to two turns of DNA within
the RecA-DNA nucleoprotein filament.
Underwinding of DNA associated with duplex-duplex pairing by RecA protein
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Hertfordshire, United Kingdom.
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