Overexpression of human peroxiredoxin 5 in subcellular compartments of Chinese hamster ovary cells: effects on cytotoxicity and DNA damage caused by peroxides.
Peroxiredoxin 5 is a mammalian thioredoxin peroxidase ubiquitously expressed in tissues. Peroxiredoxin 5 can be intracellularly localized to mitochondria, peroxisomes, the cytosol, and, to a lesser extent, the nucleus. This remarkably wide subcellular distribution compared with the five other mammalian peroxiredoxins prompted us to further investigate the antioxidant protective function of peroxiredoxin 5 in mammalian cells according to its subcellular localization. Chinese hamster ovary cells overexpressing human peroxiredoxin 5 in the cytosol, in mitochondria, or in the nucleus were established by stable transfection. Cells overexpressing peroxiredoxin 5 were exposed for 1 h to low or acute oxidative stress with exogenously added hydrogen peroxide or tert-butylhydroperoxide. Cell protection conferred by peroxiredoxin 5 was evaluated by clonogenicity and lactate dehydrogenase assays. Overexpressing peroxiredoxin 5 in either the cytosolic, mitochondrial, or nuclear compartment significantly reduced cell death, with more effective protection with overexpression of peroxiredoxin 5 in mitochondria, confirming that this organelle is a major target of peroxides. Moreover, we evaluated, with the comet assay, nuclear DNA damage induced by hydrogen peroxide or tert-butylhydroperoxide. Overexpression of peroxiredoxin 5 in the nucleus significantly decreased DNA damage induced by both peroxides. In conclusion, the present study suggests that multiple subcellular targeting of peroxiredoxin 5 in mammalian cells can be implicated in antioxidant protective mechanisms under nonpathological conditions but also during acute oxidative stress caused by peroxides occurring in pathophysiological situations.
Banmeyer
Ingrid
I
Laboratory of Cell Biology, Institut des Sciences de la Vie, Université catholique de Louvain, B-1348 Louvain-La-Neuve, Belgium.
Marchand
Cécile
C
Verhaeghe
Catherine
C
Vucic
Bénédicte
B
Rees
Jean-François
JF
Knoops
Bernard
B
D000818:Animals; D016466:CHO Cells; D002374:Catalase / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D005979:Glutathione Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D008928:Mitochondria / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics* / Q000378:metabolism*; D010545:Peroxides / Q000494:pharmacology*; D054464:Peroxiredoxins; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
D010545:Peroxides; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D010544:Peroxidases; C515974:PRDX5 protein, human; D054464:Peroxiredoxins; D002374:Catalase; D005979:Glutathione Peroxidase