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J Biol Chem, Vol. 274, Issue 22, 15345-15349, May 28, 1999
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From the Mutations of mitochondrial (mt) DNA accumulate
during normal aging. The most frequent mutation is a 4,977-base pair
deletion also called the common deletion, which is increased in
photoaged skin. Oxidative stress may play a major role in the
generation of large scale mtDNA deletions, but direct proof for this
has been elusive. We therefore assessed whether the common deletion can
be generated in vitro through UV irradiation and whether
reactive oxygen species are involved in this process. Normal human
fibroblasts were repetitively exposed to sublethal doses of UVA
radiation and assayed for the common deletion employing a
semiquantitative polymerase chain reaction technique. There was a
time/dose-dependent generation of the common deletion,
attributable to the generation of singlet oxygen, since the common
deletion was diminished when irradiating in the presence of singlet
oxygen quenchers, but increased when enhancing singlet oxygen half-life
by deuterium oxide. The induction of the common deletion by UVA
irradiation was mimicked by treatment of unirradiated cells with
singlet oxygen produced by the thermodecomposition of an endoperoxide.
These studies provide evidence for the involvement of reactive oxygen
species in the generation of aging-associated mtDNA lesions in human
cells and indicate a previously unrecognized role of singlet oxygen in
photoaging of human skin.
Clinical and Experimental Photodermatology,
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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