Elevated metals compromise repair of oxidative DNA damage via the base excision repair pathway: implications of pathologic iron overload in the brain on integrity of neuronal DNA.
Tissue-specific iron content is tightly regulated to simultaneously satisfy specialized metabolic needs and avoid cytotoxicity. In the brain, disruption of iron homeostasis may occur in acute as well as progressive injuries associated with neuronal dysfunction and death. We hypothesized that adverse effects of disrupted metal homeostasis on brain function may involve impairment of DNA repair processes. Because in the brain, the base excision repair (BER) pathway is central for handling oxidatively damaged DNA, we investigated effects of elevated iron and zinc on key BER enzymes. In vitro DNA repair assays revealed inhibitory effects of metals on BER activities, including the incision of abasic sites, 5'-flap cleavage, gap filling DNA synthesis and ligation. Using the comet assay, we showed that while metals at concentrations which inhibit BER activities in in vitro assays, did not induce direct genomic damage in cultured primary neurons, they significantly delayed repair of genomic DNA damage induced by sublethal exposure to H(2)O(2). Thus, in the brain even a mild transient metal overload, may adversely affect the DNA repair capacity and thereby compromise genomic integrity and initiate long-term deleterious sequelae including neuronal dysfunction and death.
Li
Hui
H
Department of Surgery, University of Texas Medical Branch, Galveston, Texas 77555-1220, USA.
Swiercz
Rafal
R
Englander
Ella W
EW
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D020552:Comet Assay / Q000379:methods; D004249:DNA Damage / Q000187:drug effects*; D004260:DNA Repair / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D006861:Hydrogen Peroxide / Q000633:toxicity; D007501:Iron / Q000633:toxicity; D008297:Male; D008670:Metals / Q000378:metabolism / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects* / Q000201:enzymology*; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress / Q000187:drug effects*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015032:Zinc / Q000633:toxicity
D008670:Metals; D016877:Oxidants; D006861:Hydrogen Peroxide; D007501:Iron; D015032:Zinc