Abstract: |
We report the reconstitution of the initial steps of the double-strand break-repair pathway where joint molecule formation between a duplex DNA fragment and a circular template by the combined action of RecA, RecBCD, and the single-stranded DNA binding protein provides the substrate for replication fork formation by the restart primosome and the DNA polymerase III holoenzyme. We show that PriA dictates the pathway of replication from the recombination intermediate by inhibiting a nonspecific, strand displacement DNA synthesis reaction and favoring the formation of a bona fide replication fork. Furthermore, we find that RecO and RecR significantly stimulate this recombination-directed DNA replication reaction, and that this stimulation is modulated by the presence of RecF, suggesting that the latter protein may also act as a regulator of the pathway of resolution of the recombination intermediate. |
Keywords: |
unclassified drug; dna-binding proteins; dna replication; dna synthesis; dna recombination; dna damage; dna repair; protein; dose-response relationship, drug; dna strand breakage; double stranded dna; recombination, genetic; plasmids; models, genetic; dna topoisomerase (atp hydrolysing); electrophoresis, gel, two-dimensional; replication protein a; exodeoxyribonucleases; exodeoxyribonuclease v; single stranded dna binding protein; protein pria; holoenzyme; rec a recombinases; article; dna polymerase iii holoenzyme; restart primosomal protein; reca gene
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