2001•Zhongguo yaolixue yu dulixue zazhiRequires access
Comparison of the repair percentage of oxidative damage hotspots with that of overall oxidative damage in mitochondrial DNA
Liu Li, Zhixiong Zhuang, Wen Chen, Yang Xing-fen, Qiao Zhang, Du Liutao, Qing Wei, Deng Lixia
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Abstract
In order to study whether or not the formation of mtDNA oxidative damage hotspots is related to its slow DNA repair, BRL 3A cells were exposed to 50 mmol·L -1 H 2O 2 for 30 min and then harvested at 0, 1, 4 and 24 h, respectively. The repair percentage of hotspots was analyzed by the method of LMPCR and then compared with that of overall mtDNA oxidative damage. Blot slot assay was also utilized to show the authenticity of mtDNA repair. Our results showed that at 4 and 24 h, the repair percentage of hotspots were 5.2% and 42.1%, but that of overall mtDNA oxidative repair were 76.9% and 84.1%, the repair rate of overall mtDNA oxidative damage was 1-14 folds faster than that of hospots. The results of slot blot assay showed that p53/mtDNA ratio in cellular total DNA at each time point was not significantly different. These results suggest that the formation of mtDNA oxidative damage hotspots is related to its slow repair rate, and the results of slot blot assay indicate that our repair rate can authentically describe the real conditions.