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Articles in PresS, published online ahead of print March 1, 2002
Am J Physiol Lung Cell Mol Physiol, 10.1152/ajplung.00443.2001
Submitted on November 16, 2001
Accepted on February 15, 2002
1 Cell Biology and Neuroscience, University of South Alabama, Mobile, AL, USA
2 Pharmacology, University of South Alabama, Mobile, AL, USA
3 Wells Center for Pediatric Research, Indiana University Medical School, Indianapolis, IN, USA
* To whom correspondence should be addressed. E-mail: mgillesp{at}jaguar1.usouthal.edu.
In rat cultured pulmonary arterial (PA), microvascular, and venous endothelial cells (ECs) the rate of mitochondrial (mt) DNA repair is predictive of the severity of xanthine oxidase (XO)-induced mtDNA damage and the sensitivity to XO-mediated cell death. To examine the importance of mtDNA damage and repair more directly, the impact of mitochondrial over-expression of the DNA repair enzyme, Ogg1, on XO-induced mtDNA damage and cell death was determined in PAECs. PAECs were transiently transfected with an Ogg1-mitochondrial targeting sequence construct. Mitochondrial-selective over-expression of the transgene product was confirmed microscopically by the observation that immunoreactive Ogg1 co-localized with a mitochondrial-specific tracer and, using an oligonucleotide cleavage assay, by a selective enhancement of mitochondrial Ogg1 activity. Over-expression of Ogg1 protected against both XO-induced mtDNA damage, determined by quantitative Southern analysis, and cell death as assessed by trypan blue exclusion and MTS assays. These findings show that mtDNA damage is a direct cause of cell death in XO-treated PAECs.
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