Systematic study of 2'-deoxyoxanosine formation during nitrosative deamination of 2'-deoxyguanosine under biologically relevant conditions

TOXI 115

Jose L. McFaline, jose_mc@mit.edu, Bo Pang, pangbo@mit.edu, Matthew R. Sullivan, matsul30@MIT.EDU, and Peter C. Dedon, pcdedon@mit.edu. Biological Engineering Division, Massachusetts Institute of Technology, 77 Massachusetts Avenue, NE47-293, Cambridge, MA 02139
Deamination of DNA bases occurs by both chemical and enzymatic mechanisms, including nitrosation reactions with derivatives of nitric oxide. Significant controversy surrounds the nitrosative deamination of 2¢-deoxyguanosine (dG), with evidence for the formation of both 2¢-deoxyxanthosine (dX) and 2¢-deoxyoxanosine (dO) under acidic conditions but only dX under conditions of biological pH. We have been unable to detect dO at a level of >1 per 107 nt in isolated DNA exposed to biologically relevant levels of nitrous anhydride, an NO-derived nitrosating species, or in E. coli, cultured mammalian cells or tissues from mouse models of inflammation. To test a recent model of base-pairing influences on dO formation, we have used an NO/O2 delivery system and a sensitive LC/MS-MS method to quantify the formation of dX and dO from dG as a free nucleoside and in single- and double-stranded oligonucleotides. The results shed light on the biological relevance of dO as a product of nitrosative deamination of dG.