Comparison of the molecular influences of NO-induced lesions in DNA strands on the reactivity of polynucleotide kinases, DNA ligases and DNA polymerases

초록

Nitric oxide (NO) causes DNA damage, generating xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G) and hypoxanthine (Hyp, H) from adenine (Ade, A) by nitrosative oxidation. Although these NO-induced lesions have been thought to cause mutagenic problems in cellular systems, the influence of these lesions on enzymatic functions has not yet been compared systematically. In this study, we investigated the effect of NO-induced lesions on the activities of DNA-binding/recognizing enzymes such as T4 polynucleotide kinase (T4 PNK), DNA ligases (T4 DNA ligase, Taq DNA ligase) and DNA polymerases (E. coli DNA polymerase I, Klenow fragment, T4 DNA polymerase). The phosphorylation efficiencies of T4 PNK are dependent on the base type at the 5'-end of single-stranded DNA, where Oxa congruent to Hyp congruent to Gua > Xan congruent to Ade. The enzymatic reactions efficiencies of DNA ligases or DNA polymerases were observed to be dependent on the base-pairing type bound by the enzymes, where G:C > H:C > O:C > X:C and A:T congruent to H:T > O:T > X:T. These results suggested that NO-induced lesions and their base-pairs could participate in the interaction mechanisms of the DNA-binding/recognizing enzymes in a similar manner as natural nucleobases.

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