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. 1982 Feb;79(3):744–748. doi: 10.1073/pnas.79.3.744

Identification of thymidine-5′-aldehyde at DNA strand breaks induced by neocarzinostatin chromophore

Lizzy S Kappen *, Irving H Goldberg *,†, Jerrold M Liesch ‡
PMCID: PMC345828  PMID: 16593156

Abstract

Snake venom phosphodiesterase or endonuclease S1 digestion of neocarzinostatin chromophore-treated DNA, labeled in its thymidine residues, liberates an unusual labeled nucleoside from the 5′ end of a drug-induced break. This substance, isolated by reverse-phase HPLC, possesses carbons from both the thymine and the deoxyribose moieties of thymidine in the DNA but, unlike thymidine, is readily degraded at pH 12 to thymine and a sugar fragment. The altered nucleoside was shown to contain a carbonyl group by its reduction with NaBH4 to form a substance that has the chromatographic properties of thymidine and by its reaction with various hydrazines to form the respective hydrazone derivatives; the carbonyl exists as the 5′ aldehyde as shown by its mild chemical oxidation to the carboxylic acid with simultaneous loss of the 5′ 3H. Mass spectral analysis showed a fragmentation pattern compatible with the structure thymidine-5′-aldehyde. These data indicate that the nonprotein chromophore of neocarzinostatin, in the presence of a reducing substance (2-mercaptoethanol) and molecular oxygen, selectively oxidizes the 5′ carbon of nucleosides in DNA to the aldehyde, resulting in a strand break and a DNA fragment bearing nucleoside-5′-aldehyde at its 5′ end.

Keywords: nucleoside oxidation, alkali lability, nuclease digestion, base release

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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