Duborjal Hervé, Beugnot Réjane, Mousson de Camaret Bénédicte, Issartel Jean-Paul
GENOME Express SA, 38944 Meylan, France.
Genome Res. 2002 Dec;12(12):1901-9. doi: 10.1101/gr.194102.
We have measured, by reverse transcription and real-time quantitative PCR, the steady-state levels of the mitochondrial and nuclear transcripts encoding several subunits of the human oxidative phosphorylation (OXPHOS) system, in different normal tissues (muscle, liver, trachea, and kidney) and in cultured cells (normal fibroblasts, 143B osteosarcoma cells, 143B206 rho(0) cells). Five mitochondrial transcripts and nine nuclear transcripts were assessed. The measured amounts of these OXPHOS transcripts in muscle samples corroborated data obtained by others using the serial analysis of gene expression (SAGE) method to appraise gene expression in the same type of tissue. Steady-state levels for all the transcripts were found to range over more than two orders of magnitude. Most of the time, the mitochondrial H-strand transcripts were present at higher levels than the nuclear transcripts. The mitochondrial L-strand transcript ND6 was usually present at a low level. Cultured 143B cells contained significantly reduced amounts of mitochondrial transcripts in comparison with the tissue samples. In 143B206 rho(0) cells, fully depleted of mitochondrial DNA, the levels of nuclear OXPHOS transcripts were not modified in comparison with the parental cells. This observation indicated that nuclear transcription is not coordinated with mitochondrial transcription. We also observed that in the different tissues and cells, there is a transcriptional coregulation of all the investigated nuclear genes. Nuclear OXPHOS gene expression seems to be finely regulated.
我们通过逆转录和实时定量PCR,测量了编码人类氧化磷酸化(OXPHOS)系统几个亚基的线粒体和核转录本在不同正常组织(肌肉、肝脏、气管和肾脏)以及培养细胞(正常成纤维细胞、143B骨肉瘤细胞、143B206 rho(0)细胞)中的稳态水平。评估了五个线粒体转录本和九个核转录本。在肌肉样本中测得的这些OXPHOS转录本的量,证实了其他人使用基因表达序列分析(SAGE)方法在同一类型组织中评估基因表达所获得的数据。发现所有转录本的稳态水平范围超过两个数量级。大多数情况下,线粒体H链转录本的水平高于核转录本。线粒体L链转录本ND6通常水平较低。与组织样本相比,培养的143B细胞中线粒体转录本的量显著减少。在完全缺失线粒体DNA的143B206 rho(0)细胞中,与亲代细胞相比,核OXPHOS转录本的水平没有改变。这一观察结果表明,核转录与线粒体转录不协调。我们还观察到,在不同的组织和细胞中,所有研究的核基因存在转录共调节。核OXPHOS基因表达似乎受到精细调节。