Antonicka Hana, Sasarman Florin, Kennaway Nancy G, Shoubridge Eric A
Montreal Neurological Institute, McGill University, 3801 University Street, Montreal, Quebec, Canada.
Hum Mol Genet. 2006 Jun 1;15(11):1835-46. doi: 10.1093/hmg/ddl106. Epub 2006 Apr 21.
Defects in mitochondrial translation are associated with a remarkable, but unexplained diversity of clinical phenotypes. Here we have investigated the molecular basis for tissue specificity in patients with a fatal hepatopathy due to mutations in the mitochondrial translation elongation factor EFG1. Blue-native gel electrophoresis revealed unique, tissue-specific patterns in the nature and severity of the defect. Liver was the most severely affected tissue, with less than 10% residual assembly of complexes I and IV, and a 50% decrease in complex V. Skeletal muscle showed a 50% reduction in complex I, and complexes IV and V were 20% of control. In fibroblasts, complexes I and IV were 20% of control, and there was a 40-60% reduction in complexes III and V. In contrast, except for a 50% decrease in complex IV, all complexes were near normal in heart. The severity of the defect paralleled the steady-state level of the mutant EFG1 protein, which varied from 60% of control in heart to undetectable in liver. The ratio of translation elongation factors EFTu:EFTs increased from 1:6 to 1:2 in patient heart, whereas in liver it decreased from 1:1 to 1:4. Over-expression of either EFTu or EFTs in control and patient fibroblasts produced dominant negative effects, indicating that the relative abundance of these factors is an important determinant of translation efficiency. Our results demonstrate marked differences among tissues in the organization of the mitochondrial translation system and its response to dysfunction, and explain the severe hepatopathy, but normal cardiac function in EFG1 patients.
线粒体翻译缺陷与一系列显著但原因不明的临床表型多样性相关。在此,我们研究了因线粒体翻译延伸因子EFG1突变导致致命性肝病患者组织特异性的分子基础。蓝色非变性凝胶电泳揭示了缺陷性质和严重程度方面独特的组织特异性模式。肝脏是受影响最严重的组织,复合物I和IV的残余组装量不到10%,复合物V减少50%。骨骼肌中复合物I减少50%,复合物IV和V为对照的20%。在成纤维细胞中,复合物I和IV为对照的20%,复合物III和V减少40 - 60%。相比之下,除复合物IV减少50%外,心脏中的所有复合物均接近正常。缺陷的严重程度与突变型EFG1蛋白的稳态水平平行,该水平在心脏中为对照的60%,在肝脏中无法检测到。患者心脏中翻译延伸因子EFTu:EFTs的比例从1:6增加到1:2,而在肝脏中从1:1降至1:4。在对照和成纤维细胞患者中过表达EFTu或EFTs均产生显性负效应,表明这些因子的相对丰度是翻译效率的重要决定因素。我们的结果表明,线粒体翻译系统的组织及其对功能障碍的反应在不同组织之间存在显著差异,并解释了EFG1患者严重的肝病但心脏功能正常的现象。