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Titolo:
A simple and sensitive method for in vitro quantitation of abasic sites inDNA
Autore:
Boturyn, D; Constant, JF; Defrancq, E; Lhomme, J; Barbin, A; Wild, CP;
Indirizzi:
Univ Grenoble 1, LEDSS, CNRS, UMR 5616, F-38041 Grenoble 9, France Univ Grenoble 1 Grenoble France 9 , UMR 5616, F-38041 Grenoble 9, France Int Agcy Res Canc, F-69372 Lyon, France Int Agcy Res Canc Lyon France F-69372 gcy Res Canc, F-69372 Lyon, France Univ Leeds, Mol Epidemiol Unit, Leeds LS2 9JT, W Yorkshire, England Univ Leeds Leeds W Yorkshire England LS2 9JT S2 9JT, W Yorkshire, England
Titolo Testata:
CHEMICAL RESEARCH IN TOXICOLOGY
fascicolo: 6, volume: 12, anno: 1999,
pagine: 476 - 482
SICI:
0893-228X(199906)12:6<476:ASASMF>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
DEOXYRIBONUCLEIC-ACID; APURINIC SITES; APYRIMIDINIC SITES; ESCHERICHIA-COLI; ENDONUCLEASE-III; REPAIR ENZYMES; DAMAGE; INVITRO; ASSAY; QUANTIFICATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
47
Recensione:
Indirizzi per estratti:
Indirizzo: Constant, JF Univ Grenoble 1, LEDSS, CNRS, UMR 5616, BP 53, F-38041 Grenoble 9, France Univ Grenoble 1 BP 53 Grenoble France 9 1 Grenoble 9, France
Citazione:
D. Boturyn et al., "A simple and sensitive method for in vitro quantitation of abasic sites inDNA", CHEM RES T, 12(6), 1999, pp. 476-482

Abstract

A novel method for the quantitation of abasic sites (AP sites) in DNA is described. As abasic sites can be generated by controlled thermal treatment of base-modified DNA, this method can be used for estimation of the extent of DNA damage resulting from exposure to genotoxic agents. The method involves use of probe molecules 1 and 2 that contain a fluorescent label linked to an aminooxy group which reacts specifically with the aldehydic function of the ring-opened form of abasic sites. The two fluorescent probes 1 and 2were found to react with 2-deoxyribose, a model substrate, at the optimum of PH 4.0. As spontaneous depurination occurs at low PH, the reactions withabasic DNA were carried out at neutral PH with an excess concentration of the probes. Studies with alkylated, depurinated calf thymus DNA showed thatthe method is selective and quantitative. Good correlations were found between the level of 7-methylguanine (7-MeGua), generated in vitro in DNA by the methylating agent dimethyl sulfate, and the amount of AP sites as determined by the method presented here. In addition, similar correlations were found when the assay was used to detect abasic sites in DNA isolated from rats treated with carcinogenic alkylating agents. In each case, the level of abasic sites, as expected, is slightly higher than the level of 7-MeGua which is known to represent about 70% of the total modifications of DNA following exposure to the methylating agent. This method may be useful not only in experimental settings but also in studies of DNA damage in humans resulting from chemotherapy or exposure to environmental agents.

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Documento generato il 05/12/20 alle ore 14:13:51