Detection of alkylation damage in human lymphocyte DNA with the comet assay

• Autorzy: Collins A. R. , Dusinska M. , Horska A.
• Języki publikacji: EN

Abstrakty

EN

The enzyme 3-methyladenine DNA glycosylase II (AlkA) is abacterial repair enzyme that acts preferentially at 3-methyladenine residues in DNA, releasing the damaged base. The resulting baseless sugars are alkali-labile, and under the conditions of the alkaline comet assay (single cell gel electrophoresis) they appear as DNA strand breaks. AlkA is not lesion-specific, but has a low activity even with undamaged bases. We have tested the enzyme at different concentrations to find conditions that maxi­mise detection of alkylated bases with minimal attack on normal, undamaged DNA. AlkA detects damage in the DNA of cells treated with low concentrations of methyl methanesulphonate. We also find low background levels of alkylated bases in normal human lymphocytes.

Słowa kluczowe

EN

alkylation damage; 4,6-diamidino-2-phenylindole; methyl methanesulphonate; formamidopyrimidine DNA glycosylase; enzyme; lymphocyte; human lymphocyte; DNA damage; 3-methyladenine DNA glycosylase II; comet assay; DNA; detection

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 2001
• Tom: 48
• Numer: 3
• Opis fizyczny: p.611-614,fig.

Twórcy

autor

Collins A. R.

• Rowett Research Institute, Greemburn Road, Bucksburn, Aberdeen AB21 9SB, Scotland, U.K.

autor

Dusinska M.

autor

Horska A.

Bibliografia

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• 3.Collins, A.R., Duthie, S.J. & Dobson, V.L. (1993) Direct enzymic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis 14, 1733-1735.
• 4.Collins, A.R., Duthie, S.J., Fillion, L., Gedik, C.M., Vaughan, N. & Wood, S.G. (1997a) Oxidative DNA damage in human cells: The influence of antioxidants and DNA repair. Biochem. Soc. Trans. 25, 326-331.
• 5.Collins, A.R., Mitchell, D.L., Zunino, A., de Wit, J. & Busch, D. (1997b) UV-sensitive rodent mutant cell lines of complementation groups 6 and 8 differ phenotypically from their human counterparts. Environ. Mol. Mutagen. 29, 152-160.
• 6.Dusinska, M. & Collins, A. (1996) Detection of oxidised purines and UV-induced photoproducts in DNA of single cells, by inclusion of lesion-specific enzymes in the comet assay. Altern. Lab. Anim. 24, 405-411.
• 7.Dusinska, M., Lietava, J., Olmedilla, B., Raslova, K., Southon, S. & Collins, A.R. (1999) Indicators of oxidative stress, antioxidants and human health; in Antioxidants in Human Health (Basu, T.K., Temple, N.J. & Garg, M.L., eds.) pp. 411-422, CABI Publishing, Oxford, U.K.
• 8.Lindahl, T. (1993) Instability and decay of the primary structure of DNA. Nature 362, 709-715.