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RNA facilitates RecA-mediated DNA pairing and strand transfer between molecules bearing limited regions of homology
Authors:Hidehito Kotani  Markus W Germann  Alex Andrus  Ravi Vinayak  Bashar Mullah and Eric B Kmiec
Institution:(1) Department of Pharmacology, Jefferson Cancer Center, Thomas Jefferson University, 233 S. 10th Street, 19107 Philadelphia, PA, USA;(2) Division of Perlsin-Elman Group, Applied Biosystems, Inc., 850 Lincoln Drive, 94404 Foster City, CA, USA
Abstract:The RecA protein ofEscherichia coli catalyzes homologous pairing and strand exchange between a wide range of molecules showing nucleotide sequence complementarity, including a linear duplex and a single-stranded DNA molecule. We demonstrate that RecA can promote formation of joint molecules when the duplex contains an RNA/DNA hairpin and a single-stranded circle serves as the pairing partner. A chimeric RNA/DNA hairpin can be used to form stable joint molecules with as little as 15 bases of shared homology as long as the RNA stretch contains complementarity to the circle. The joint molecule bears some resemblance to a triple helical structure composed of RNA residues surrounded by two DNA strands which are in a parallel orientation. Evidence is presented that supports the notion that short stretches of RNA can be used in homologous pairing reactions at lengths below that required for DNA-DNA heteroduplex formation.
Keywords:RNA  RecA protein  Homologous recombination
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