Enzymatic Incorporation of Modified Purine Nucleotides in DNA

A series of nucleotide analogues, with a hypoxanthine base moiety (8-aminohypoxanthine, 1-methyl-8-aminohypoxanthine, and 8-oxohypoxanthine), together with 5-methylisocytosine were tested as potential pairing partners of N8-glyco sylated nucleotides with an 8-azaguanine or 8-aza-9-deaza guanine base moiety by using DNA polymerases (incorporation studies). The best results were obtained with the 5-methyl isocytosine nucleotide followed by the 1-methyl-8-aminohypo xanthine nucleotide. The experiments demonstrated that small differences in the structure (8-azaguanine versus 8-aza-9-deazaguanine) might lead to significant differences in recognition efficiency and selectivity, base pairing by Hoogsteen recognition at the polymerase level is possible, 8-aza-9-deazaguanine represents a self-complementary base pair, and a correlation exists between in vitro incorporation studies and in vivo recognition by natural bases in Escherichia coli, but this recognition is not absolute (exceptions were observed).status: publishe