Abstract
Addition of thiol compounds containing an anionic group to the 3'-terminal unsaturated sugar of the 5' fragment obtained from an oligonucleotide containing an AP site cleaved by beta-elimination, can be followed by gel electrophoresis. The technique enables to distinguish between two mechanisms of cleavage of the C3'-O-P bond 3' to an AP site: hydrolysis or beta-elimination. Addition of thiols to the double-bond of the 3'-terminal sugar resulting from beta-elimination prevents a subsequent delta-elimination. The interpretation of the action of enzymes that start by nicking 3' to AP sites must take into account the presence or absence of thiols in the reaction medium. In living cells, thiols might influence the pathways followed by the repair processes of AP site-containing DNA.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bailly V., Verly W. G. Escherichia coli endonuclease III is not an endonuclease but a beta-elimination catalyst. Biochem J. 1987 Mar 1;242(2):565–572. doi: 10.1042/bj2420565. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bailly V., Verly W. G. Possible roles of beta-elimination and delta-elimination reactions in the repair of DNA containing AP (apurinic/apyrimidinic) sites in mammalian cells. Biochem J. 1988 Jul 15;253(2):553–559. doi: 10.1042/bj2530553. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bailly V., Verly W. G. The excision of AP sites by the 3'-5' exonuclease activity of the Klenow fragment of Escherichia coli DNA polymerase I. FEBS Lett. 1984 Dec 10;178(2):223–227. doi: 10.1016/0014-5793(84)80605-5. [DOI] [PubMed] [Google Scholar]
- César R., Verly W. G. The apurinic/apyrimidinic endodeoxyribonuclease of rat-liver chromatin. Eur J Biochem. 1983 Jan 1;129(3):509–517. doi: 10.1111/j.1432-1033.1983.tb07077.x. [DOI] [PubMed] [Google Scholar]
- Gossard F., Verly W. G. Properties of the main endonuclease specific for apurinic sites of Escherichia coli (endonuclease VI). Mechanism of apurinic site excision from DNA. Eur J Biochem. 1978 Jan 16;82(2):321–332. doi: 10.1111/j.1432-1033.1978.tb12026.x. [DOI] [PubMed] [Google Scholar]
- Mosbaugh D. W., Linn S. Further characterization of human fibroblast apurinic/apyrimidinic DNA endonucleases. The definition of two mechanistic classes of enzyme. J Biol Chem. 1980 Dec 25;255(24):11743–11752. [PubMed] [Google Scholar]
- Verly W. G., Paquette Y., Thibodeau L. Nuclease for DNA apurinic sites may be involved in the maintenance of DNA in normal cells. Nat New Biol. 1973 Jul 18;244(133):67–69. doi: 10.1038/newbio244067a0. [DOI] [PubMed] [Google Scholar]