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J. Biol. Chem., Vol. 261, Issue 28, 13087-13096, 10, 1986
DK Gonda and CM Radding
recA protein promotes the homologous pairing of single strands with duplex
DNA by polymerizing on the single strands to make presynaptic nucleoprotein
filaments which are polyvalent with respect to duplex DNA and which
consequently form large networks or coaggregates when duplex DNA is added.
Previous work has shown that efficient homologous pairing occurs within
these networks. In the experiments described here, we observed that the
length of the duplex DNA determined the stability of coaggregates, their
steady state level, and the yield of joint molecules. Correspondingly,
heterologous duplex DNA when preincubated with presynaptic filaments
excluded subsequently added homologous duplex DNA from coaggregates and
inhibited homologous pairing; the extents of exclusion and inhibition were
determined by the length of the heterologous duplex DNA. On the other hand,
long heterologous duplex DNA when added together with short homologous
duplex DNA was capable of stimulating the absorption of the homologous
molecules into coaggregates and increasing the rate of homologous pairing.
In reactions involving short duplex molecules, polyamines exerted
comparable effects on coaggregation and homologous pairing. We conclude
that coaggregates are instrumental in homologous pairing, that they
constitute distinct domains that are responsible for the processive or
first order character of the pairing reaction, and that they act by
concentrating DNA and facilitating diffusion.
The mechanism of the search for homology promoted by recA protein. Facilitated diffusion within nucleoprotein networks
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