RecA protein-mediated irreversible fixation of an oligodeoxyribonucleotide to specific site in DNA

RecA protein can polymerize on an oligodeoxyribonucleotide to form a filament that finds its homologous sequence in double-stranded DNA. When such an oligonucleotide is linked to psoralen, a photoactivatable DNA intercalator, it irreversibly binds to the homologous site in double-stranded DNA as a result of psoralen photoadduct formation at thymidines. The relative efficiency of specific vs. non-specific binding of an oligonucleotide depended upon the ratio of psoralenated oligonucleotide to total DNA. Na+ ions at concentrations greater than 50 mM eliminated specific binding. Under optimal conditions, the probability of binding of an 80-mer oligonucleotide to a specific site was > 10(5) times greater than that of binding to any single non-specific site. Under the conditions described, RecA-mediated photoadduction was equally efficient with superhelical and linear double-stranded DNA.