Establishing the role of the atypical kinase ADCK3 in mitochondrial metabolism

确定非典型激酶 ADCK3 在线粒体代谢中的作用

• 批准号: 8900321
• 负责人: David J Pagliarini
• 金额: $ 7.42万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8900321
• 关键词: Address; Affect; Anabolism; Animal Model; Architecture; Ataxia; Biochemical; Biochemistry; Biological Process; Biology; Cataloging; Catalogs; Cell Culture Techniques; Cell Line; Cerebellar Ataxia; Data; Disease; Electron Transport; Environment; Estrogen receptor positive; Etiology; Eukaryotic Cell; Extensive Stage; Family; Fibroblasts; Foundations; Functional disorder; Future; Gene Mutation; Genes; Genetic; Glioblastoma; Goals; Health; Human; In Vitro; Inborn Errors of Metabolism; Inner mitochondrial membrane; Knockout Mice; Lipids; Malignant Neoplasms; Mammalian Cell; Mammary Neoplasms; Mass Spectrum Analysis; Measurement; Membrane; Metabolism; Mitochondria; Mitochondrial Diseases; Modeling; Molecular; Mus; Mutate; Mutation; Organelles; Orthologous Gene; Outcome; Pathway interactions; Patients; Phenotype; Phospholipids; Phosphotransferases; Physiological; Property; Proteins; Proteome; RNA Interference; Reporting; Research Infrastructure; Research Personnel; Resources; Respiratory Chain; Role; Saccharomyces cerevisiae; Signal Transduction; Structure; System; Techniques; Testing; Ubiquinone; Work; Workplace; Yeasts; base; cell type; chemical genetics; design; human disease; in vitro Assay; insight; lipid metabolism; member; metabolomics; mitochondrial dysfunction; mouse model; novel; preference; programs; protein function; protein metabolite; tool

DESCRIPTION (provided by applicant): Mitochondria are centers of metabolism and signaling whose function is essential to all but a few eukaryotic cell types. General dysfunction of these organelles is implicated in a wide range of inborn errors of metabolism, and in an increasing number of common human diseases. Elucidation of the biochemical functions of these disease-related proteins has become a bottleneck in understanding mitochondrial pathophysiology. This proposal initially focuses on one such protein, ADCK3, a predicted kinase with a poorly understood connection to the biosynthesis of coenzyme Q, an integral part of the mitochondrial electron transport chain. Mutations in this protein can cause a form of cerebellar ataxia, one of numerous mitochondrial respiratory chain disorders. More broadly, ADCK3 is part of a large and ancient family of predicted atypical kinases-the UbiB family-that has no structural or functional characterization reported to date. This proposal will establish the fundamental enzymatic properties of this archetypal member of the UbiB family, identify its direct endogenous substrate(s), and test the hypothesis that it functions to remodel the protein infrastructure of th inner mitochondrial membrane through alterations in the local phospholipid environment. Leveraging the depth and breadth of expertise available through the investigators of this program project, the proposed work incorporates a diverse range of tools to address these goals, including: a robust, high mass-accuracy lipidomics platform and yeast, mammalian cell culture and mouse knockout models of ADCK3 deficiency. As the characterization of ADCK3 progresses, the approaches and techniques established here will subsequently be applied to the other ADCK kinases that are generally conserved from yeast through metazoans. These kinases include four in human mitochondria (ADCK1, 2, 4 and 5), which have now been associated both with lipid metabolism and with specific cancers, including glioblastoma multiforme (GBM) and estrogen receptor-positive (ER+) breast tumors. Thus, in addition to the direct insights that will be generate regarding the specific function of ADCK3, this work sets the stage for an extensive analysis of a unique mitochondrial kinase family of mounting biomedical importance.

描述（由申请人提供）：线粒体是代谢和信号传导的中心，其功能对于除少数真核细胞类型之外的所有真核细胞类型都是必需的。这些细胞器的一般功能障碍与广泛的先天性代谢缺陷以及越来越多的常见人类疾病有关。阐明这些疾病相关蛋白的生化功能已成为理解线粒体病理生理学的瓶颈。该建议最初集中在一个这样的蛋白质，ADCK 3，预测激酶与辅酶Q的生物合成，线粒体电子传递链的一个组成部分知之甚少。这种蛋白质的突变会导致小脑共济失调，这是众多线粒体呼吸链疾病之一。更广泛地说，ADCK 3是一个庞大而古老的预测非典型激酶家族的一部分-UbiB家族-迄今为止没有报道结构或功能特征。该提案将建立UbiB家族的这种原型成员的基本酶性质，鉴定其直接内源性底物，并测试其功能是通过改变局部磷脂环境来重塑线粒体内膜的蛋白质基础结构的假设。利用该计划项目的研究人员提供的专业知识的深度和广度，拟议的工作采用了多种工具来实现这些目标，包括：一个强大的，高质量精度的脂质组学平台和酵母，哺乳动物细胞培养和ADCK 3缺陷的小鼠敲除模型。随着ADCK 3的表征进展，本文建立的方法和技术随后将应用于通常从酵母到后生动物保守的其他ADCK激酶。这些激酶包括人类线粒体中的四种（ADCK 1，2，4和5），它们现在与脂质代谢和特定癌症（包括多形性胶质母细胞瘤（GBM）和雌激素受体阳性（ER+）乳腺肿瘤）相关。因此，除了将产生关于ADCK 3的特定功能的直接见解之外，这项工作还为广泛分析具有越来越重要的生物医学意义的独特线粒体激酶家族奠定了基础。