Regulation of C. elegans SKN-1/Nrf activity by the unfolded protein response

通过未折叠蛋白反应调节线虫 SKN-1/Nrf 活性

基本信息

批准号：
8233869
负责人：
T Keith Blackwell
金额：
$ 28.19万
依托单位：
JOSLIN DIABETES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-30 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8233869
关键词：
Address Affect Animal Model Animals Biological Biological Process Caenorhabditis elegans Cell physiology Cells Cellular Stress Complex Data Defense Mechanisms Diabetes Mellitus Disease Endoderm Endoplasmic Reticulum Fatty acid glycerol esters Gene Expression Gene Targeting Gene Transfer Techniques Genes Genetic Genome Growth Hepatitis Hepatocyte Homeostasis Insulin Insulin-Like Growth Factor I Integral Membrane Protein Intestines Liver Failure Longevity Longevity Pathway Mammalian Cell Mammals Mediating Modeling Molecular Genetics Molecular Profiling Nerve Degeneration Neurons OmpR protein Organism Oxidation-Reduction Oxidative Stress Pathway interactions Play Process Processed Genes Protein Biosynthesis Protein Isoforms Proteins RNA Interference Regulation Resistance Role Screening procedure Secretory Cell Series Signal Pathway Signal Transduction Sirolimus Stress System Testing Tissues Toxin Transgenic Organisms Work Xenobiotics base biological adaptation to stress chromatin immunoprecipitation endoplasmic reticulum stress in vivo insight meetings multicatalytic endopeptidase complex novel protein folding research study response secretory protein sensor tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Many diverse diseases arise from an excess of unfolded proteins in the endoplasmic reticulum (ER stress). ER stress induces the unfolded protein response (UPR), in which numerous protective genes are activated. It is critical to understand how the UPR defends against ER stress, and how UPR signaling affects other cellular processes. While three canonical UPR transcription factors have been identified, we have determined in C. elegans that the transcription factor SKN-1 (mammalian Nrf1/2/3) is also central to the UPR. SKN-1 has a conserved function in oxidative stress defense, and is important in longevity. We have shown that SKN-1 is regulated by the UPR, and is needed for ER stress to activate core UPR regulators, as well as downstream effector genes. Surprisingly, UPR signaling is required for SKN-1 to respond to oxidative stress, which suggests that ER signaling may broadly influence cellular stress defenses, and may regulate SKN-1 in the insulin-IGF-1-like signaling (IIS) and TOR (target of rapamycin) pathways. Using the powerful C. elegans model, this project will investigate: (1) how and in what tissues SKN-1 defends against ER stress, and how IIS and TOR might affect ER stress resistance through SKN-1, (2) how SKN-1 is regulated by the UPR during ER stress, oxidative stress, and in the context of the IIS and TOR pathways, and (3) what genes and biological processes are controlled directly by SKN-1 during the UPR. Transgenic, genetic, molecular, cell biological, and chromatin immunoprecipitation (ChIP) approaches will be used to investigate how SKN-1 functions are regulated by the UPR and contribute to ER stress resistance in vivo. A proven RNAi screening strategy will identify new regulators of SKN-1 during ER stress, and high-throughput sequencing combined with expression profiling and ChIP will be employed to identify genes and processes regulated directly by SKN-1. These studies will provide important and novel insights into how the ER defends itself against stress, and will substantially alter paradigms for understanding ER-based signaling and how it influences other cellular defense mechanisms. PUBLIC HEALTH RELEVANCE: Many proteins are synthesized in the endoplasmic reticulum (ER). High levels of misfolded ER proteins (ER stress) cause or exacerbate disease states as diverse as diabetes, hepatitis, and neurodegeneration. This project will use a model organism (C. elegans) to determine how a regulator of diverse cellular defenses protects against ER stress, and how signaling from the ER controls this and other functions of this regulator.

描述（由申请人提供）：许多不同的疾病是由内质网中过量的未折叠蛋白（ER 应激）引起的。内质网应激会诱导未折叠蛋白反应（UPR），其中许多保护基因被激活。了解 UPR 如何防御 ER 应激以及 UPR 信号如何影响其他细胞过程至关重要。虽然已经鉴定出三种典型的 UPR 转录因子，但我们在秀丽隐杆线虫中确定转录因子 SKN-1（哺乳动物 Nrf1/2/3）也是 UPR 的核心。 SKN-1 在氧化应激防御中具有保守的功能，对长寿很重要。我们已经证明 SKN-1 受 UPR 调节，并且是 ER 应激激活核心 UPR 调节因子以及下游效应基因所必需的。令人惊讶的是，UPR 信号是 SKN-1 响应氧化应激所必需的，这表明 ER 信号可能广泛影响细胞应激防御，并可能在胰岛素-IGF-1 样信号 (IIS) 和 TOR 中调节 SKN-1。雷帕霉素的靶标）途径。使用强大的线虫模型，该项目将研究：(1) SKN-1 如何以及在哪些组织中抵御 ER 应激，以及 IIS 和 TOR 如何通过 SKN-1 影响 ER 应激抵抗，(2) SKN-1 如何1 在 ER 应激、氧化应激以及 IIS 和 TOR 通路背景下受 UPR 调节，以及 (3) 在 UPR 期间哪些基因和生物过程直接受 SKN-1 控制。转基因、遗传、分子、细胞生物学和染色质免疫沉淀 (ChIP) 方法将用于研究 SKN-1 功能如何受 UPR 调节并有助于体内内质网应激抵抗。经过验证的 RNAi 筛选策略将识别 ER 应激期间 SKN-1 的新调节因子，并且将采用高通量测序结合表达谱和 ChIP 来识别 SKN-1 直接调节的基因和过程。这些研究将为内质网如何防御应激提供重要而新颖的见解，并将大大改变理解基于内质网的信号传导及其如何影响其他细胞防御机制的范式。公共卫生相关性：许多蛋白质在内质网 (ER) 中合成。高水平的错误折叠 ER 蛋白（ER 应激）会导致或加剧糖尿病、肝炎和神经退行性疾病等多种疾病。该项目将使用模型生物（线虫）来确定多种细胞防御的调节剂如何防止内质网应激，以及来自内质网的信号如何控制该调节剂的这一功能和其他功能。