Mitochondrial DNA damage and a hypoxic response are induced by CoCl(2) in rat neuronal PC12 cells.
Generation of reactive oxygen species (ROS) and activation of a transcriptional program that mimics the hypoxic response have been documented in cultured cells in the presence of cobalt chloride. We found that in the presence of hypoxia-mimicking concentrations of CoCl(2), mitochondrial but not nuclear DNA damage is induced in rat neuronal, PC12 cells. To our knowledge, this is the first documentation of induction of mitochondrial DNA (mtDNA) damage under these conditions. Likewise, we provide the first evidence for elevation of MYH, the mammalian homolog of the Escherichia coli MutY DNA glycosylase, in mammalian cells. Recently, the human MYH was implicated in repair of oxidative DNA damage and shown to carry a mitochondrial localization sequence. Here, an induction of mtDNA damage and a time-dependent increase in the MYH level were detected with exposure of cells to 100 microM CoCl(2). In addition, the levels of proteins involved in cellular responses to hypoxia, ROS and nuclear DNA damage; hypoxia-inducible factor 1alpha(HIF-1alpha), p53, p21 and PCNA were also modulated temporally. Earlier studies suggested that the mtDNA is a primary target for oxidative damage. Our findings extend these observations and suggest that activation of DNA repair processes is associated with the presence of mtDNA damage.
Wang
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G
Department of Surgery and Shriners Hospitals for Children and Sealy Center for Molecular Science, The University of Texas Medical Branch, Galveston, TX 77555, USA.
Hazra
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TK
Mitra
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Lee
H M
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Englander
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EW
D000818:Animals; D016587:Antimutagenic Agents / Q000494:pharmacology; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology*; D003035:Cobalt / Q000494:pharmacology*; D004249:DNA Damage; D045647:DNA Glycosylases; D004272:DNA, Mitochondrial / Q000187:drug effects* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007700:Kinetics; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects*; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D051381:Rats
D016587:Antimutagenic Agents; D004272:DNA, Mitochondrial; D003035:Cobalt; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; C061738:mutY adenine glycosylase; C018021:cobaltous chloride