Enhanced mtDNA repair capacity protects pulmonary artery endothelial cells from oxidant-mediated death

Allison W. Dobson, Valentina Grishko, Susan P. Ledoux, Mark Kelley, Glenn L. Wilson, Mark N. Gillespie

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

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, we determined the impact of mitochondrial overexpression of the DNA repair enzyme, Ogg1, on XO-induced mtDNA damage and cell death in PAECs. PAECs were transiently transfected with an Ogg1-mitochondrial targeting sequence construct. Mitochondria-selective overexpression of the transgene product was confirmed microscopically by the observation that immunoreactive Ogg1 colocalized with a mitochondria-specific tracer and, with an oligonucleotide cleavage assay, by a selective enhancement of mitochondrial Ogg1 activity. Overexpression 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.

Original languageEnglish
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume283
Issue number1 27-1
StatePublished - 2002

Fingerprint

Mitochondrial DNA
Oxidants
Pulmonary Artery
Xanthine Oxidase
Endothelial Cells
Cell Death
Mitochondria
DNA Repair Enzymes
Trypan Blue
Transgenes
Oligonucleotides
DNA Repair
Cause of Death
Lung

Keywords

  • Cytotoxicity
  • Mitochondrial deoxyribonucleic acid
  • Ogg1
  • Xanthine oxidase

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

Cite this

Enhanced mtDNA repair capacity protects pulmonary artery endothelial cells from oxidant-mediated death. / Dobson, Allison W.; Grishko, Valentina; Ledoux, Susan P.; Kelley, Mark; Wilson, Glenn L.; Gillespie, Mark N.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 283, No. 1 27-1, 2002.

Research output: Contribution to journalArticle

Dobson, Allison W. ; Grishko, Valentina ; Ledoux, Susan P. ; Kelley, Mark ; Wilson, Glenn L. ; Gillespie, Mark N. / Enhanced mtDNA repair capacity protects pulmonary artery endothelial cells from oxidant-mediated death. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2002 ; Vol. 283, No. 1 27-1.
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