DESIGN AND SYNTHESIS OF DOUBLE-STRANDED OLIGONUCLEOTIDES CONTAINING REACTIVE ACYLPHOSPHATE INTERNUCLEOTIDE GROUPS

DNA duplex and dumbbells containing chemically active acylphosphate in ternucleotide groups were synthesized. To obtain these compounds the c hemical ligation method was used. The acylphosphate group was inserted into a DNA duplex and dumbbells as a result of template-directed cond ensation of 5'-phosphate and especially introduced 3'-carboxy groups o f oligonucleotides, 1-Ethyl-3-(3'-dimethylaminopropyl)carbodiimide (ED C) was used as a condensing agent. Oligonucleotides containing a carbo xy group mere obtained by the interaction of their 3'-phosphate with g lycine methyl ester under the action of EDC, followed by ester hydroly sis, The yields of acylphosphate-containing double-stranded oligonucle otides achieved 15-25% depending on the structure of their precursors. It was shown that these compounds are acylating agents and are effici ently cleaved in near-physiological conditions under the action of eth ylenediamine or N-methylimidazole. These results indicate that doubles tranded oligonucleotides carrying acylphosphate internucleotide groups could constitute new crosslinking reagents for affinity modification of DNA recognizing proteins. (C) 1998 Federation of European Biochemic al Societies.