DESIGN OF NEW REAGENTS ON THE BASE OF DNA DUPLEXES FOR IRREVERSIBLE INHIBITION OF TRANSCRIPTION FACTOR NF-KAPPA-B

The main purpose of the present work is to search for the optimal desi gn of a DNA duplex containing an active group for crosslinking and irr eversible inhibition of the transcription factor NF-kappa B. Modified DNA duplexes with an identical nucleotide sequence but different inter nucleotide phosphates replaced by the trisubstituted pyrophosphate int ernucleotide group were synthesized. Crosslinking of the human NF-kapp a B p50 subunit with the modified DNA duplexes was carried out. It was shown that only four modified duplexes crosslinked with the NF-kappa B p50 subunit. The specificity of these reactions was confirmed. A pos ition of the phosphate in the NF-kappa B recognition site was found wh ere replacement on the active trisubstituted pyrophosphate group resul ted in a 50% yield of crosslinking. The fact that DNA duplexes contain ing the trisubstituted pyrophosphate group specifically react with the NF-kappa B p50 subunit in the Escherichia coli total lysate supports the idea that such modified DNA can be used as high specific inhibitor s for DNA-recognizing proteins.