Nucleic Acids Research, Vol 26, Issue 9 2184-2191, Copyright © 1998 by Oxford University Press
M Jaiswal, LJ Lipinski, VA Bohr and SJ Mazur
To investigate the repair of oxidative damage in DNA, we have established
an in vitro assay utilizing human lymphoblastoid whole cell extracts and
plasmid DNA damaged by exposure to methylene blue and visible light. This
treatment has been shown to produce predominantly 7-
hydro-8-oxodeoxyguanosine (8-oxodG) in double-stranded DNA at low levels of
modification. DNA containing 1. 6 lesions per plasmid is substrate for
efficient repair synthesis by cell extracts. The incorporation of dGMP is
2.7 +/- 0.5 times greater than the incorporation of dCMP, indicating an
average repair patch of 3-4 nucleotides. Damage-specific nicking occurs
within 15 min, while resynthesis is slower. The incorporation of dGMP
increases linearly, while the incorporation of dCMP exhibits a distinct
lag. Extracts from xeroderma pigmentosum (XP) complementation groups A and
B exhibit 25 and 40%, respectively, of the incorporation of dCMP compared
with normal extracts, but extracts from an XP-D cell line exhibit twice the
activity. These data suggest that the efficient repair of 8-oxodG lesions
observed in human cell extracts involves more than one pathway of base
excision repair.
ARTICLES
Efficient in vitro repair of 7-hydro-8-oxodeoxyguanosine by human cell extracts: involvement of multiple pathways
Laboratory of Molecular Genetics, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
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