de Souza-Pinto N C, Hogue B A, Bohr V A
Laboratory of Molecular Genetics, NIA, National Institutes of Health, Baltimore, MD 21224, USA.
Free Radic Biol Med. 2001 Apr 15;30(8):916-23. doi: 10.1016/s0891-5849(01)00483-x.
8-oxo-deoxyguanosine (8-oxodG) is one of the major DNA lesions formed upon oxidative attack of DNA. It is a mutagenic adduct that has been associated with pathological states such as cancer and aging. Base excision repair (BER) is the main pathway for the repair of 8-oxodG. There is a great deal of interest in the question about age-associated accumulation of this DNA lesion and its intracellular distribution, particularly with respect to mitochondrial or nuclear localization. We have previously shown that 8-oxodG-incision activity increases with age in rat mitochondria obtained from both liver and heart. In this study, we have investigated the age-associated changes in DNA repair activities in both mitochondrial and nuclear extracts obtained from mouse liver. We observed that 8-oxodG incision activity of mitochondrial extracts increases significantly with age, from 13.4 + or - 2.2 fmoles of oligomer/100 microg of protein/16 h at 6 to 18.6 + or - 4.9 at 14 and 23.7 + or - 3.8 at 23 months of age. In contrast, the nuclear 8-oxodG incision activity showed no significant change with age, and in fact slightly decreased from 11.8 + or - 3 fmoles/50 microg of protein/2 h at 6 months to 9.7 + or - 0.8 at 14 months. Uracil DNA glycosylase and endonuclease G activities did not change with age in nucleus or mitochondria. Our results show that the repair of 8-oxodG is regulated differently in nucleus and mitochondria during the aging process. The specific increase in 8-oxodG-incision activity in mitochondria, rather than a general up-regulation of DNA metabolizing enzymes in those organelles, suggests that this pathway may be up regulated during aging in mice.
8-氧代脱氧鸟苷(8-oxodG)是DNA受到氧化攻击后形成的主要DNA损伤之一。它是一种诱变加合物,与癌症和衰老等病理状态有关。碱基切除修复(BER)是修复8-oxodG的主要途径。人们对这种DNA损伤与年龄相关的积累及其细胞内分布问题,特别是关于线粒体或细胞核定位,有着极大的兴趣。我们之前已经表明,从肝脏和心脏获取的大鼠线粒体中,8-oxodG切割活性会随着年龄增长而增加。在本研究中,我们调查了从小鼠肝脏获取的线粒体和细胞核提取物中与年龄相关的DNA修复活性变化。我们观察到,线粒体提取物的8-oxodG切割活性随着年龄显著增加,从6个月时的13.4±2.2飞摩尔寡聚物/100微克蛋白质/16小时,增加到14个月时的18.6±4.9,以及23个月时的23.7±3.8。相比之下,细胞核的8-oxodG切割活性并未随年龄发生显著变化,实际上从6个月时的11.8±3飞摩尔/50微克蛋白质/2小时略微下降到14个月时的9.7±0.8。尿嘧啶DNA糖基化酶和核酸内切酶G的活性在细胞核或线粒体中均未随年龄发生变化。我们的结果表明,在衰老过程中,8-oxodG在细胞核和线粒体中的修复调控方式不同。线粒体中8-oxodG切割活性的特异性增加,而非这些细胞器中DNA代谢酶的普遍上调,表明该途径在小鼠衰老过程中可能被上调。