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 design of a DNA duplex containing an active group for crosslinking and irreversible inhibition of the transcription factor NF-kappa B. Modified DNA duplexes with an identical nucleotide sequence but different internucleotide phosphates replaced by the trisubstituted pyrophosphate internucleotide group were synthesized. Crosslinking of the human NF-kappa 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 position of the phosphate in the NF-kappa B recognition site was found where replacement on the active trisubstituted pyrophosphate group resulted in a 50% yield of crosslinking. The fact that DNA duplexes containing 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 inhibitors for DNA-recognizing proteins.
Kozlov
I A
IA
Department of Chemistry, Moscow State University, Russia.
Kubareva
E A
EA
Ivanovskaya
M G
MG
Shabarova
Z A
ZA
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents; D018360:Crystallography, X-Ray; D004247:DNA / Q000378:metabolism*; D011756:Diphosphates; D015195:Drug Design; D004926:Escherichia coli; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry
D003432:Cross-Linking Reagents; D011756:Diphosphates; D046911:Macromolecular Substances; D016328:NF-kappa B; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D004247:DNA