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[2'-Deoxynucleoside-5'-triphosphate, modified via the carbohydrate and triphosphate residues, in the DNA synthesis
T A Rozovskaia1, A V Mishchenko, N B Tarusova
1Institute of Chemistry, National University of Mexico, Mexico.
Abstract:
We have investigated the substrate properties of deoxyribonucleoside 5'-triphosphate analogues, modified in the carbohydrate and triphosphate moieties, in DNA synthesis catalyzed by different DNA polymerases and reverse transcriptases. It was shown that (3'-azido-2',3'-dideoxythymidine-5'-O-methylenephosphonate) diphosphate, (3'-azido-2',3'-dideoxythymidine 5'-phosphate) dibromomethylenediphosphonate, (3'-azido-2',3'-dideoxythymidine 5'-phosphate) phosphonoacetate terminate DNA synthesis catalyzed by reverse transcriptases. (2'-Deoxythymidine 5'-phosphate) phosphonoacetate displays substrate properties for DNA polymerase beta, different reverse transcriptases, terminal deoxynucleotidyl transferase, but not for DNA polymerase alpha, Klenow's fragment DNA polymerase I. The Km value for this substance in DNA synthesis reactions catalyzed by reverse transcriptases was two of orders magnitude higher than that for native 2'-deoxythymidine 5'-triphosphate.