FASTKD2 is an RNA-binding protein required for mitochondrial RNA processing and translation.
FASTKD2 is an RNA-binding protein required for mitochondrial RNA processing and translation.
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DOI：
10.1261/rna.052365.115
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发表时间：
2015-11
期刊：
影响因子：
--
通讯作者：
Hentze MW
中科院分区：
文献类型：
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作者：
Popow J;Alleaume AM;Curk T;Schwarzl T;Sauer S;Hentze MW
线粒体 RNA 加工是所有真核生物中电子传递链成分合成的重要步骤，但线粒体 RNA 生物合成和调控的几个方面尚未得到充分了解。 RNA 相互作用组捕获鉴定了几种具有非典型 RNA 结合结构的疾病相关 RNA 结合蛋白 (RBP)，包括 FASTK（FAS 激活的丝氨酸/苏氨酸激酶）蛋白家族的所有六个成员。这些新指定的 FASTK RBP 之一（FASTKD2）内的突变会导致一种罕见的孟德尔线粒体脑肌病。为了研究 FASTKD2 的 RNA 结合是否会导致疾病表型，我们通过 iCLIP 鉴定了 FASTKD2 的 RNA 靶标。 FASTKD2 与一组确定的线粒体转录物相互作用，包括 16S 核糖体 RNA (RNR2) 和 NADH 脱氢酶亚基 6 (ND6) 信使 RNA。 CRISPR 介导的 FASTKD2 缺失会导致编码呼吸复合物 I 亚基的 RNR2 和 ND6 mRNA 的异常加工和表达。FASTKD2 缺陷细胞的代谢表型显示细胞呼吸受损，所有呼吸复合物的活性降低。这项工作通过基因组、分子和代谢分析的结合，确定了以前未表征的、与疾病相关的 RNA 结合蛋白 FASTKD2 分子网络的关键方面。
Mitochondrial RNA processing is an essential step for the synthesis of the components of the electron transport chain in all eukaryotic organisms, yet several aspects of mitochondrial RNA biogenesis and regulation are not sufficiently understood. RNA interactome capture identified several disease-relevant RNA-binding proteins (RBPs) with noncanonical RNA-binding architectures, including all six members of the FASTK (FAS-activated serine/threonine kinase) family of proteins. A mutation within one of these newly assigned FASTK RBPs, FASTKD2, causes a rare form of Mendelian mitochondrial encephalomyopathy. To investigate whether RNA binding of FASTKD2 contributes to the disease phenotype, we identified the RNA targets of FASTKD2 by iCLIP. FASTKD2 interacts with a defined set of mitochondrial transcripts including 16S ribosomal RNA (RNR2) and NADH dehydrogenase subunit 6 (ND6) messenger RNA. CRISPR-mediated deletion of FASTKD2 leads to aberrant processing and expression of RNR2 and ND6 mRNA that encodes a subunit of the respiratory complex I. Metabolic phenotyping of FASTKD2-deficient cells reveals impaired cellular respiration with reduced activities of all respiratory complexes. This work identifies key aspects of the molecular network of a previously uncharacterized, disease-relevant RNA-binding protein, FASTKD2, by a combination of genomic, molecular, and metabolic analyses.