Nucleic Acids Research, Vol 24, Issue 8 1389-1394, Copyright © 1996 by Oxford University Press
P Jaruga and M Dizdaroglu
Oxidative DNA damage is the most frequent type of damage encountered by
aerobic cells and may play an important role in biological processes such
as mutagenesis, carcinogenesis and aging in humans. Oxidative damage
generates a myriad of modifications in DNA. We investigated the cellular
repair of DNA base damage products in DNA of cultured human lymphoblast
cells, which were exposed to oxidative stress by H2O2. This DNA-damaging
agent is known to cause base modifications in genomic DNA of mammalian
cells [Dizdaroglu, M., Nackerdien, Z., Chao, B.-C., Gajewski, E. and Rao,
G. (1991) Arch. Biochem. Biophys. 285, 388-390]. Following treatment with
H2O2, the culture medium was freed from H2O2 and cells were incubated for
time periods ranging from 10 min to 6 h. DNA was isolated from control
cells, hydrogen peroxide-treated cells and cells incubated after H2O2
exposure. DNA samples were analyzed by gas chromatography/isotope-dilution
mass spectrometry. Eleven modified bases were identified and quantified.
The results showed a significant formation of these DNA base products upon
H2O2-treatment of cells. Subsequent incubation of cells caused a
time-dependent excision of these products from cellular DNA. The cell
viability did not change significantly by various treatments. There were
distinct differences between the kinetics of excision of individual
products. The observed excisions were attributed to DNA repair in cells.
The rate of repair of purine lesions was slower than that of pyrimidine
lesions. Most of the identified products are known to possess various
premutagenic properties. The results of this work may contribute to the
understanding of the cellular repair of oxidative DNA damage in human and
other mammalian cells.
ARTICLES
Repair of products of oxidative DNA base damage in human cells
Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaitersburg, MD 20899, USA.
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