Stable three-stranded DNA made by RecA protein.
When RecA protein, in the form of a nucleoprotein filament containing circular single-stranded DNA (plus strand only), reacts with homologous linear duplex DNA, a directional transfer ensues of a strand from the duplex DNA to the nucleoprotein filament, resulting in the displacement of the linear plus strand in the 5' to 3' direction. The initial homologous synapsis, however, can occur at either end of the duplex DNA, or anywhere in between, and when homology is restricted to different regions of the duplex DNA, the joint molecules that form in each region show striking differences in stability upon deproteinization: distal joints greater than proximal joints much greater than medial joints. In the deproteinized distal joints, which are thermostable, 2000 nucleotide residues of the circular plus strand are resistant to P1 nuclease; both strands of the original duplex DNA remain resistant to P1 nuclease, and the potentially displaceable linear plus strand, which has a 3' homologous end, remains resistant to Escherichia coli exonuclease I. These observations suggest that RecA protein promotes homologous pairing and strand exchange via long three-stranded DNA intermediates and, moreover, that, once formed, such triplex structures in natural DNA are stable even when RecA protein has been removed.
Rao
B J
BJ
Department of Human Genetics, Yale University School of Medicine, New Haven, CT 06510.
Dutreix
M
M
Radding
C M
CM
D004247:DNA / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004926:Escherichia coli; D005090:Exodeoxyribonucleases / Q000494:pharmacology; D006358:Hot Temperature; D066298:In Vitro Techniques; D009690:Nucleic Acid Conformation; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
D004277:DNA, Single-Stranded; D004247:DNA; D011938:Rec A Recombinases; D005090:Exodeoxyribonucleases; C028450:exodeoxyribonuclease I; D015719:Single-Strand Specific DNA and RNA Endonucleases