Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
Proc Natl Acad Sci U S A. 2011 May 3;108(18):7437-42. doi: 10.1073/pnas.1100084108. Epub 2011 Apr 18.
Ataxia with oculomotor apraxia 1 is caused by mutation in the APTX gene, which encodes the DNA strand-break repair protein aprataxin. Aprataxin exhibits homology to the histidine triad superfamily of nucleotide hydrolases and transferases and removes 5'-adenylate groups from DNA that arise from aborted ligation reactions. We report herein that aprataxin localizes to mitochondria in human cells and we identify an N-terminal amino acid sequence that targets certain isoforms of the protein to this intracellular compartment. We also show that transcripts encoding this unique N-terminal stretch are expressed in the human brain, with highest production in the cerebellum. Depletion of aprataxin in human SH-SY5Y neuroblastoma cells and primary skeletal muscle myoblasts results in mitochondrial dysfunction, which is revealed by reduced citrate synthase activity and mtDNA copy number. Moreover, mtDNA, not nuclear DNA, was found to have higher levels of background DNA damage on aprataxin knockdown, suggesting a direct role for the enzyme in mtDNA processing.
眼动运动不能伴共济失调 1 是由 APTX 基因突变引起的,该基因编码 DNA 链断裂修复蛋白 aprataxin。Aprataxin 与组氨酸三联体超家族的核苷酸水解酶和转移酶具有同源性,可从连接反应失败产生的 DNA 中去除 5'-腺苷酸基团。我们在此报告 aprataxin 定位于人细胞中的线粒体,并且鉴定出靶向该蛋白的某些同工型的 N 末端氨基酸序列到该细胞内隔室。我们还表明,编码这种独特的 N 末端延伸的转录本在人脑表达,在小脑中有最高的产量。在人 SH-SY5Y 神经母细胞瘤细胞和原代骨骼肌成肌细胞中耗尽 aprataxin 导致线粒体功能障碍,这通过柠檬酸合酶活性和 mtDNA 拷贝数降低来揭示。此外,在 aprataxin 敲低时发现 mtDNA(而非核 DNA)具有更高水平的背景 DNA 损伤,这表明该酶在 mtDNA 加工中具有直接作用。