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）：许多多种疾病是由于内质网中未折叠的蛋白质过量引起的（ER应激）。 ER应力诱导展开的蛋白质反应（UPR），其中许多保护基因被激活。了解UPR如何防御ER应力以及UPR信号如何影响其他细胞过程至关重要。尽管已经确定了三个规范UPR转录因子，但我们在秀丽隐杆线虫中确定了转录因子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，并且在寿命中很重要。我们已经表明，SKN-1受UPR的调节，而ER应力需要激活核心UPR调节剂以及下游效应子基因。令人惊讶的是，SKN-1需要响应氧化应激，这表明ER信号传导可能会广泛影响细胞应力防御，并且可能调节胰岛素-IGF-1样信号传导（IIS）和TOR（Rapamycin）途径的SKN-1。使用强大的秀丽隐杆线虫模型，该项目将调查：（1）SKN-1如何以及在哪些组织中进行防御应力，以及IIS和TOR如何通过SKN-1来影响ER压力的抗压力，（2）（2）在ER压力，氧化压力，氧化压力，IIS和TOR途径以及IIS和TOR PATH的上下文中，SKN-1如何受到UPR的调节，以及在什么基因和（3）的上下文中，以及在skn中直接通过Skn进行了skn。转基因，遗传，分子，细胞生物学和染色质免疫沉淀（CHIP）方法将用于研究如何通过UPR调节SKN-1功能，并在体内有助于ER抗应力抗性。经过验证的RNAI筛选策略将在ER应力期间确定SKN-1的新调节剂，并且将采用高通量测序与表达分析和CHIP结合起来，以识别由SKN-1直接调节的基因和过程。这些研究将为ER如何防御压力提供重要和新颖的见解，并将大大改变理解基于ER的信号的范式以及它如何影响其他细胞防御机制。公共卫生相关性：许多蛋白质是在内质网中合成的（ER）。高水平的错误折叠的ER蛋白（ER应激）引起或加剧疾病状态，如糖尿病，肝炎和神经变性。该项目将使用模型生物体（秀丽隐杆线虫）来确定各种细胞防御的调节剂如何保护ER应力，以及来自ER的信号如何控制该调节器的此功能和其他功能。