Abstract |
8-Nitro-2'-deoxyguanosine (8-nitrodG) is a relatively unstable, mutagenic lesion of DNA that is increasingly believed to be associated with tissue inflammation. Due to the lability of the glycosidic bond, 8-nitrodG cannot be incorporated into oligodeoxynucleotides (ODNs) by chemical DNA synthesis and thus very little is known about its physicochemical properties and base-pairing preferences. Here we describe the synthesis of 8-nitro-2'-O-methylguanosine, a ribonucleoside analogue of this lesion, which is sufficiently stable to be incorporated into ODNs. Physicochemical studies demonstrated that 8-nitro-2'-O-methylguanosine adopts a syn conformation about the glycosidic bond; thermal melting studies and molecular modelling suggest a relatively stable syn-8-nitroG·anti-G base pair. Interestingly, when this lesion analogue was placed in a primer-template system, extension of the primer by either avian myeloblastosis virus reverse transcriptase (AMV-RT) or human DNA polymerase β (pol β), was significantly impaired, but where incorporation opposite 8-nitroguanine did occur, pol β showed a 2:1 preference to insert dA over dC, while AMV-RT incorporated predominantly dC. The fact that no 8-nitroG·G base pairing is seen in the primer extension products suggests that the polymerases may discriminate against this pairing system on the basis of its poor geometric match to a Watson-Crick pair.
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Authors | Inder Bhamra, Patricia Compagnone-Post, Ian A O'Neil, Lesley A Iwanejko, Andrew D Bates, Richard Cosstick |
Journal | Nucleic acids research
(Nucleic Acids Res)
Vol. 40
Issue 21
Pg. 11126-38
(Nov 2012)
ISSN: 1362-4962 [Electronic] England |
PMID | 22965127
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- 8-nitro-2'-O-methylguanosine
- 8-nitroguanine
- Oligodeoxyribonucleotides
- Guanosine
- Guanine
- DNA
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Topics |
- Base Pairing
- DNA
(biosynthesis)
- DNA Damage
- Guanine
(analogs & derivatives, chemistry)
- Guanosine
(analogs & derivatives, chemical synthesis, chemistry)
- Hydrolysis
- Mutagenesis
- Mutation
- Oligodeoxyribonucleotides
(chemical synthesis, chemistry)
- Templates, Genetic
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