Nuclear-mitochondrial co-regulation during mitochondrial biogenesis

线粒体生物发生过程中核线粒体的共同调节

基本信息

批准号：
9289152
负责人：
Lee Stirling Churchman
金额：
$ 33.45万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-15 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9289152
关键词：
Acute Address Biochemical Biogenesis Biology Carbon Cardiac Myocytes Cells Chemicals Complex Data Defect Degenerative Disorder Diabetes Mellitus Disease Engineering Equilibrium Functional disorder Gene Expression Gene Expression Process Gene Expression Regulation Genes Genetic Transcription Genome Goals Human In Vitro Lead Light Maintenance Malignant Neoplasms Mammalian Cell Maps Measures Mitochondria Mitochondrial Diseases Monitor Nerve Degeneration Neurodegenerative Disorders Nuclear Oxidative Phosphorylation Parkinson Disease Physiological Processes Process Production Protein Biosynthesis Public Health RNA Regulation Regulator Genes Research Resolution Ribosomes Role Saccharomyces cerevisiae Source Stress Testing Transcriptional Regulation Translational Regulation Translations Up-Regulation Work Yeasts biological adaptation to stress environmental chemical induced pluripotent stem cell insight mitochondrial dysfunction mitochondrial genome prevent programs response ribosome profiling transcriptome sequencing

项目摘要

PROJECT SUMMARY/ABSTRACT Defects in the assembly and maintenance of mitochondrial oxidative phosphorylation (OXPHOS) machinery lead to a range of degenerative illnesses, including diabetes, cancer, and neurodegenerative diseases. OXPHOS complexes are encoded on both the nuclear and mitochondrial genomes, so their biogenesis requires the precise coordination of gene regulatory mechanisms across genomes. To this end, mitochondrial biogenesis and stress response programs involve the simultaneous transcriptional upregulation of nuclear- encoded OXPHOS genes and mitochondrial gene expression factors. These transcriptional programs are thought to facilitate nuclear-mitochondrial balance, where mitochondrial-encoded OXPHOS subunits assemble in stoichiometric ratios with their nuclear-encoded counterparts. We recently observed in Saccharomyces cerevisiae that transcription regulation of nuclear-encoded and mitochondrial-encoded OXPHOS subunits are not coordinated during carbon source adaptation. Instead, the cell synchronizes the translational regulation of OXPHOS subunits across compartments. Whether synchronized translation is a widespread response remains unknown. The goal of this proposal is to determine how mitochondrial and nuclear genomes are co-regulated, particularly during protein synthesis, throughout mitochondrial biogenesis and stress response programs. As an extension of our recent work in yeast, Aim 1 will investigate whether synchronous translation programs occur across a range of environmental and mitochondrial stress adaptation programs. We will also determine how the synchronous translation regulation occurs by determining the role of key mitochondrial translation regulators in the dynamic regulation of OXPHOS genes during carbon source adaptation. This will be done using our mitochondrial ribosome profiling approach and cytosolic ribosome profiling to measure protein synthesis and RNA-seq to measure global transcription. In Aims 2 and 3, we extend our studies to human cells through re-engineering ribosome profiling to robustly capture human mitochondrial translation. To test the approach, we will investigate how a putative translation activator, TACO1, impacts human mitochondrial translation. We will investigate nuclear-mitochondrial co-regulation after acute mitochondrial stress induced by chemical inhibition of OXPHOS complexes, mimicking OXPHOS dysfunction in disease processes. Finally, we will investigate mitochondrial and nuclear gene expression programs during the in vitro differentiation of iPS cells to cardiomyocytes, when extensive mitochondrial biogenesis occurs. Determining the regulation and the extent of nuclear-mitochondrial co-regulation will provide critical insight towards understanding how imbalanced production of OXPHOS subunits transpires in disease states.

项目概要/摘要线粒体氧化磷酸化（OXPHOS）机制的组装和维护缺陷导致一系列退行性疾病，包括糖尿病、癌症和神经退行性疾病。 OXPHOS 复合物在核基因组和线粒体基因组上编码，因此它们的生物发生需要跨基因组的基因调控机制的精确协调。为此，线粒体生物发生和应激反应程序涉及核的同时转录上调编码 OXPHOS 基因和线粒体基因表达因子。这些转录程序是被认为有助于促进核线粒体平衡，线粒体编码的 OXPHOS 亚基在此聚集与其核编码对应物成化学计量比。我们最近在酵母菌中观察到酿酒酵母中核编码和线粒体编码的 OXPHOS 亚基的转录调控是碳源适应过程中不协调。相反，细胞同步翻译调节 OXPHOS 亚基跨区室。同步翻译是否得到广泛响应仍然存在未知。该提案的目标是确定线粒体和核基因组如何共同调节，特别是在蛋白质合成过程中、整个线粒体生物发生和应激反应程序中。作为作为我们最近在酵母方面的工作的延伸，目标 1 将研究同步翻译程序是否发生在一系列环境和线粒体应激适应计划中。我们还将确定通过确定关键线粒体翻译的作用，同步翻译调节是如何发生的碳源适应过程中 OXPHOS 基因动态调节的调节因子。这将完成使用我们的线粒体核糖体分析方法和胞质核糖体分析来测量蛋白质合成和 RNA-seq 来测量全局转录。在目标 2 和 3 中，我们将研究扩展到人类通过重新设计核糖体分析来强有力地捕获人类线粒体翻译。测试方法，我们将研究假定的翻译激活剂 TACO1 如何影响人类线粒体翻译。我们将研究急性线粒体应激后核线粒体的共同调节 OXPHOS 复合物的化学抑制，模拟疾病过程中 OXPHOS 功能障碍。最后，我们将研究 iPS 体外分化过程中的线粒体和核基因表达程序当广泛的线粒体生物发生发生时，细胞转变为心肌细胞。确定监管和核线粒体共同调节的程度将为理解如何进行提供重要的见解 OXPHOS 亚基的产生不平衡在疾病状态下发生。