Biophysical Stability and Enzymatic Recognition of Oxanine in Dna

Abstract

Oxanine (Oxa), which is one of the major products generated from guanine by nitrosative oxidation and is as long-lived as Gua in DNA, has been thought to be one of the major causes for NO-induced DNA damage. In the present study, using several synthetic Oxa-containing oligodeoxynucleotides, biophysical stability and enzymatic recognition of Oxa was investigated in DNA strands. It was found that Oxa did not mediate marked distortion in the whole DNA structure although Oxa pairing with 4 normal bases decreased thermal stability of the DNA duplexes compared to Gua:Cyt base pair. Regarding the responses of the DNA-relevant enzymes to Oxa, it was determined that Oxa was recognized as Gua except that DNA polymerases incorporated Thy as well as Cyt opposite Oxa. These results imply that Oxa tends to behave as a kind of naturally occurring base, Gua and therefore, would be involved in the genotoxic and cytotoxic threats of NO in cellular system.

Keywords:

• Oxanine
• oligodeoxynucleotide
• thermal stability
• DNA-relevant enzymes

This work is supported by the Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan to K. M. (No. 18350083), and by the Grant-in-Aid for regional science and technology promotion “Kyoto Nanotechnology Cluster” project from the MEXT, Japan. This work was also supported by CREST of the Japan Science and Technology Agency.