Bi-allelic Mutations in NDUFA6 Establish Its Role in Early-Onset Isolated Mitochondrial Complex I Deficiency.
Bi-allelic Mutations in NDUFA6 Establish Its Role in Early-Onset Isolated Mitochondrial Complex I Deficiency.
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DOI：
10.1016/j.ajhg.2018.08.013
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发表时间：
2018-10-04
影响因子：
9.8
通讯作者：
Taylor RW
中科院分区：
文献类型：
--
作者：
Alston CL;Heidler J;Dibley MG;Kremer LS;Taylor LS;Fratter C;French CE;Glasgow RIC;Feichtinger RG;Delon I;Pagnamenta AT;Dolling H;Lemonde H;Aiton N;Bjørnstad A;Henneke L;Gärtner J;Thiele H;Tauchmannova K;Quaghebeur G;Houstek J;Sperl W;Raymond FL;Prokisch H;Mayr JA;McFarland R;Poulton J;Ryan MT;Wittig I;Henneke M;Taylor RW
孤立的复合物I缺乏症是儿科线粒体疾病中观察到的常见生化表型，并且通常是由于致病性变体影响编码复合物I结构亚基或组装因子的p565基因之一而引起的。这种遗传异质性意味着将下一代测序技术应用于未确诊的队列，是基因诊断和基因-疾病关联的催化剂。我们描述了四个不相关的儿童的临床和分子遗传学研究，他们表现为神经放射学检查结果和/或乳酸水平升高，高度提示潜在的线粒体诊断。下一代测序鉴定了NDUFA 6中的双等位基因变体，其编码形成Q模块的一部分的含有15 kDa LYR基序的复合物I亚基。使用受试者的成纤维细胞系进行的功能研究证明了复合物I的组装缺陷，并通过基于质谱的复合物组分析对其进行了详细表征。这证实了掺入的NDUFA 6的显著减少和其他Q-模块亚基（包括NDUFAB 1、NDUFA 7和NDUFA 12）的伴随减少。受试者成纤维细胞的慢病毒转导显示复合物I的正常化。这些数据也支持超复合物的形成，其中的1830 kDa的复合物I中间体（由P-和Q-模块组成）是在复杂的组装复合物III和IV全酶，尽管缺乏N-模块。有趣的是，RNA测序数据提供的证据表明，共有RefSeq登录号与临床相关组织中的主要转录物不对应，这促使对NDUFA 6 RefSeq转录物进行修订，不仅强调了彻底的变体解释的重要性，而且还强调了评估适当的转录物以进行分析的重要性。
Isolated complex I deficiency is a common biochemical phenotype observed in pediatric mitochondrial disease and often arises as a consequence of pathogenic variants affecting one of the ∼65 genes encoding the complex I structural subunits or assembly factors. Such genetic heterogeneity means that application of next-generation sequencing technologies to undiagnosed cohorts has been a catalyst for genetic diagnosis and gene-disease associations. We describe the clinical and molecular genetic investigations of four unrelated children who presented with neuroradiological findings and/or elevated lactate levels, highly suggestive of an underlying mitochondrial diagnosis. Next-generation sequencing identified bi-allelic variants in NDUFA6, encoding a 15 kDa LYR-motif-containing complex I subunit that forms part of the Q-module. Functional investigations using subjects’ fibroblast cell lines demonstrated complex I assembly defects, which were characterized in detail by mass-spectrometry-based complexome profiling. This confirmed a marked reduction in incorporated NDUFA6 and a concomitant reduction in other Q-module subunits, including NDUFAB1, NDUFA7, and NDUFA12. Lentiviral transduction of subjects’ fibroblasts showed normalization of complex I. These data also support supercomplex formation, whereby the ∼830 kDa complex I intermediate (consisting of the P- and Q-modules) is in complex with assembled complex III and IV holoenzymes despite lacking the N-module. Interestingly, RNA-sequencing data provided evidence that the consensus RefSeq accession number does not correspond to the predominant transcript in clinically relevant tissues, prompting revision of the NDUFA6 RefSeq transcript and highlighting not only the importance of thorough variant interpretation but also the assessment of appropriate transcripts for analysis.
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