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

Andrew Collins | Zendy; Mária Dušinská | Zendy; Alexandra Horská | Zendy

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

Author(s) -

Andrew Collins,

Mária Dušinská,

Alexandra Horská

Publication year - 2001

Publication title -

acta biochimica polonica

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.452

H-Index - 78

eISSN - 1734-154X

pISSN - 0001-527X

DOI - 10.18388/abp.2001_3895

Subject(s) - dna glycosylase , dna , dna damage , comet assay , chemistry , enzyme , biochemistry , alkylation , microbiology and biotechnology , dna repair , biology , catalysis

The enzyme 3-methyladenine DNA glycosylase II (AlkA) is a bacterial 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 no t lesion-specific, but has a low activity even w ith undamagedbases. 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.

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