Institute of Human Genetics, Technische Universität München, Munich, Germany.
Mol Genet Metab. 2011 Jun;103(2):161-6. doi: 10.1016/j.ymgme.2011.03.004. Epub 2011 Mar 11.
Mitochondrial complex I deficiency is a frequent biochemical condition, causing about one third of respiratory chain disorders. Partly due to the large number of genes necessary for its assembly and function only a small proportion of complex I deficiencies are yet confirmed at the molecular genetic level. Now, next generation sequencing approaches are applied to close the gap between biochemical definition and molecular diagnosis. Nevertheless such approaches result in a long list of novel rare single nucleotide variants. Identifying the causative mutations still remains challenging. Here we describe the identification and functional confirmation of novel NDUFS1 mutations using a cellular rescue-assay. Patient-derived complex I-defective fibroblast cell lines were transduced with wild type and mutant NDUFS1-cDNA and subsequently analyzed on the functional and protein level. We established the pathogenic nature of identified rare variants in four out of five disease alleles. This approach is a valuable add-on in disease genetics and it allows the analysis of the functional consequences of genetic variants in metabolic disorders.
线粒体复合物 I 缺陷是一种常见的生化状况，导致约三分之一的呼吸链紊乱。部分由于其组装和功能所需的大量基因，只有一小部分复合物 I 缺陷在分子遗传水平上得到证实。现在，下一代测序方法被应用于缩小生化定义和分子诊断之间的差距。然而，这些方法会产生一长串新的罕见单核苷酸变异。确定致病突变仍然具有挑战性。在这里，我们使用细胞拯救实验描述了新型 NDUFS1 突变的鉴定和功能确认。用野生型和突变型 NDUFS1-cDNA 转导患者来源的复合物 I 缺陷成纤维细胞系，并在功能和蛋白水平上进行分析。我们确定了五个疾病等位基因中的四个中鉴定出的罕见变异的致病性。这种方法是疾病遗传学的一个有价值的补充，它允许分析代谢紊乱中遗传变异的功能后果。
