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

基本信息

批准号：
8765976
负责人：
David J Pagliarini
金额：
$ 31.79万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8765976
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 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 public health relevance tool

项目摘要

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多种先天性代谢错误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。该提案最初关注的是这样一种蛋白质 ADCK3，这是一种预测的激酶，与辅酶 Q（线粒体电子传递链的组成部分）的生物合成之间的联系知之甚少。这种蛋白质的突变可导致某种形式的小脑共济失调，这是众多线粒体呼吸链疾病之一。更广泛地说，ADCK3 是一个古老的预测非典型激酶家族（UbiB 家族）的一部分，迄今为止尚未有结构或功能特征报道。该提案将建立 UbiB 家族原型成员的基本酶特性，鉴定其直接内源底物，并测试其通过改变局部磷脂环境来重塑线粒体内膜蛋白质基础设施的假设。利用该计划项目研究人员提供的专业知识的深度和广度，拟议的工作结合了多种工具来实现这些目标，包括：强大的、高质量准确的脂质组学平台和酵母、哺乳动物细胞培养和小鼠基因敲除ADCK3 缺陷模型。随着 ADCK3 表征的进展，本文建立的方法和技术将随后应用于通常从酵母到后生动物中保守的其他 ADCK 激酶。这些激酶包括人类线粒体中的四种激酶（ADCK1、2、4 和 5），它们现已与脂质代谢和特定癌症相关，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。因此，除了对 ADCK3 的特定功能产生直接见解之外，这项工作还为广泛分析具有日益重要的生物医学重要性的独特线粒体激酶家族奠定了基础。