Montreal Neurological Institute and Department of Human Genetics, McGill University, Montreal H3A 2B4, Quebec, Canada.
Am J Hum Genet. 2010 Jul 9;87(1):115-22. doi: 10.1016/j.ajhg.2010.06.004.
We investigated the genetic basis for a global and uniform decrease in mitochondrial translation in fibroblasts from patients in two unrelated pedigrees who developed Leigh syndrome, optic atrophy, and ophthalmoplegia. Analysis of the assembly of the oxidative phosphorylation complexes showed severe decreases of complexes I, IV, and V and a smaller decrease in complex III. The steady-state levels of mitochondrial mRNAs, tRNAs, and rRNAs were not reduced, nor were those of the mitochondrial translation elongation factors or the protein components of the mitochondrial ribosome. Using homozygosity mapping, we identified a 1 bp deletion in C12orf65 in one patient, and DNA sequence analysis showed a different 1 bp deletion in the second patient. Both mutations predict the same premature stop codon. C12orf65 belongs to a family of four mitochondrial class I peptide release factors, which also includes mtRF1a, mtRF1, and Ict1, all characterized by the presence of a GGQ motif at the active site. However, C12orf65 does not exhibit peptidyl-tRNA hydrolase activity in an in vitro assay with bacterial ribosomes. We suggest that it might play a role in recycling abortive peptidyl-tRNA species, released from the ribosome during the elongation phase of translation.
我们研究了两个无关联家系中 Leigh 综合征、视神经萎缩和眼肌麻痹患者成纤维细胞中线粒体翻译普遍下降的遗传基础。氧化磷酸化复合物组装分析显示复合物 I、IV 和 V 严重减少,复合物 III 减少较少。线粒体 mRNA、tRNA 和 rRNA 的稳态水平没有降低,线粒体翻译延伸因子或线粒体核糖体的蛋白质成分也没有降低。我们使用纯合性作图在一位患者中鉴定出 C12orf65 中的 1 bp 缺失,DNA 序列分析显示第二位患者存在不同的 1 bp 缺失。这两种突变均预测出相同的过早终止密码子。C12orf65 属于线粒体 I 类肽释放因子家族的四个成员之一,该家族还包括 mtRF1a、mtRF1 和 Ict1,它们都在活性位点具有 GGQ 基序。然而,C12orf65 在体外与细菌核糖体的试验中不表现出肽酰-tRNA 水解酶活性。我们推测,它可能在翻译延长阶段从核糖体上释放出的无活性肽酰-tRNA 物种的再循环中发挥作用。