Mitchondria-Nuclear Crosstalk in Hypoxic Gene Induction

缺氧基因诱导中的线粒体-核串扰

基本信息

批准号：
7921850
负责人：
ROBERT Oliver POYTON
金额：
$ 20.51万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7921850
关键词：
Address Aerobic Affect Aging Anemia Anoxia Blood Vessels Cell Nucleus Cells Cellular Structures Central Sleep Apnea Disease Eukaryota Event Exposure to Failure Genes Genetic Genomics Goals Grant Human Hypercapnic respiratory failure Hypoxia Inflammatory Response Lead Link Liver Mitochondria Mammalian Cell Metabolism Methodology Methods Mitochondria Modeling Monitor Myocardial Infarction Nitrates Nitric Oxide Nitrites Nuclear Organism Oxygen Pathway interactions Physiological Play Process Production Proteins Proteomics RNA Interference Rattus Reactive Oxygen Species Reporting Respiratory Chain Retinal Diseases Role Signal Pathway Signal Transduction Signal Transduction Pathway Syndrome Tissues Tyrosine Yeasts angiogenesis base experience gene induction genetic manipulation inhibitor/antagonist knock-down nitration nitrosative stress null mutation oxidation protein function research study respiratory sensory system transcription factor tumor growth

项目摘要

DESCRIPTION (provided by applicant): The ability of cells, tissues, and organisms to sense and respond to changes in oxygen levels is often crucial to their survival. Cellular adaptation to hypoxia is a good example of this. Intermittent episodes of hypoxia are associated with a variety of human patho-physiological states (e.g., sleep apnea, central hypoventilation syndrome, and vascular occlusion); these can alter metabolism, induce angiogenesis, and affect inflammatory responses. Several studies have reported that adaptation to hypoxia requires the induction of a large number of nuclear genes and that the mitochondrial respiratory chain plays an important role in the induction of some of these hypoxic genes in mammalian and yeast cells. The overall goal of our studies is to understand the role of the mitochondrion in this process. These studies combine proteomic, genomic, and genetic strategies to analyze hypoxic gene induction in yeast. In addition, parallel studies will be done with mammalian cells in culture. Aim 1 will use gene profiling methods to identify hypoxic nuclear genes that are under the control of the mitochondrial respiratory chain in yeast. Aim 2 will focus on the mitochondrial - dependent hypoxic gene induction pathway itself. Genetic manipulation will be used to identify and order yeast genes that are essential for the induction of mitochondrial-dependent hypoxic nuclear genes. Aim 3 will evaluate the importance of mitochondrially-generated reactive oxygen species and nitric oxide in yeast hypoxic gene induction, with emphasis on understanding the role of protein tyrosine nitration and protein carbonylation in hypoxic signaling. Aim 4 focuses on hypoxic gene induction in mammalian cells. It addresses two questions that are controversial or incompletely explored. First, it will use methodologies that have been successful in yeast to ask if mammalian cells experience transient oxidative or nitrosative stress when exposed to hypoxia or anoxia and, if so, identify proteins carbonylated or tyrosine nitrated under anoxic or hypoxic conditions. Second, it will use in RNAi knock down experiments to assess the importance of these carbonylated or nitrated proteins in hypoxic signaling in mammalian cells. The failure of cells to respond properly to hypoxia can lead to a variety of pathological states (e.g., anemia, myocardial infarction, retinopathy, and the growth of tumors). An understanding of hypoxic signaling pathways may lead to new therapies for these diseases and may help with our understanding of human aging.

描述（由申请人提供）：细胞，组织和生物的感知和响应氧气水平变化的能力通常对它们的生存至关重要。细胞适应缺氧是一个很好的例子。缺氧的间歇性发作与多种人类病态生理状态有关（例如，睡眠呼吸暂停，中央中央低文化综合征和血管闭塞）；这些可以改变新陈代谢，诱导血管生成并影响炎症反应。几项研究报告说，适应缺氧需要诱导大量的核基因，线粒体呼吸链在诱导哺乳动物和酵母细胞中某些缺氧基因的诱导中起着重要作用。我们研究的总体目标是了解线粒体在此过程中的作用。这些研究结合了蛋白质组学，基因组和遗传策略，以分析酵母中的低氧基因诱导。此外，将对培养物中的哺乳动物细胞进行平行研究。 AIM 1将使用基因分析方法来鉴定酵母中线粒体呼吸链控制下的低氧核基因。 AIM 2将集中于线粒体 - 依赖性低氧基因诱导途径本身。遗传操作将用于识别和订购酵母基因，这对于诱导线粒体依赖性低氧核基因至关重要。 AIM 3将评估线粒体生成的活性氧和一氧化氮在酵母低氧基因诱导中的重要性，重点是理解蛋白酪氨酸硝化和蛋白质羰基在低氧信号中的作用。 AIM 4专注于哺乳动物细胞中缺氧基因诱导。它解决了两个有争议或不完全探讨的问题。首先，它将使用在酵母中成功的方法来询问哺乳动物细胞在暴露于缺氧或缺氧的情况下是否会经历瞬时氧化或硝化应激，如果是的，则在缺氧或低氧条件下识别碳碳碳碳碳碳碳碳碳中硝酸或酪氨酸。其次，它将用于RNAi敲除实验，以评估哺乳动物细胞中低氧信号传导中这些羰基或硝化蛋白的重要性。细胞无法正确反应缺氧会导致多种病理状态（例如贫血，心肌梗死，视网膜病和肿瘤的生长）。对低氧信号通路的理解可能会导致这些疾病的新疗法，并可能有助于我们对人类衰老的理解。