Nucleic Acids Research, Vol 26, Issue 2 662-668, Copyright © 1998 by Oxford University Press
RM Anson, DL Croteau, RH Stierum, C Filburn, R Parsell and VA Bohr
Photoactivated methylene blue was used to damage purified DNA and the
mitochondrial DNA (mtDNA) of human fibroblasts in culture. The primary
product of this reaction is the DNA lesion 7-hydro-8-oxo-deoxyguanosine
(8-oxo-dG). The DNA damage was quantitated using Escherichia coli
formamidopyrimidine DNA glycosylase (Fpg) in a gene-specific damage and
repair assay. Assay conditions were refined to give incision at all
enzyme-sensitive sites with minimal non-specific cutting. Cultured
fibroblasts were exposed to photoactivated methylene blue under conditions
that would produce an average of three oxidative lesions per
double-stranded mitochondrial genome. Within 9 h, 47% of this damage had
been removed by the cells. This removal was due to repair rather than to
replication, cell loss or degradation of damaged genomes. The rate of
repair was measured in both DNA strands of the frequently transcribed
ribosomal region of the mitochondrial genome and in both strands of the
non-ribosomal region. Fpg-sensitive alkali-resistant oxidative base damage
was efficiently removed from human mtDNA with no differences in the rate of
repair between strands or between two different regions of the genome that
differ substantially with regard to transcriptional activity.
ARTICLES
Homogenous repair of singlet oxygen-induced DNA damage in differentially transcribed regions and strands of human mitochondrial DNA
Laboratory of Molecular Genetics and Laboratory of Biological Chemistry, National Institute on Aging, Baltimore, MD, USA.
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