Comparative Biology Elucidation of Environmental Pathways and Susceptibility

环境途径和敏感性的比较生物学阐明

基本信息

批准号：
7629806
负责人：
SAMUEL T LAMITINA
金额：
$ 37.41万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7629806
关键词：
Address Aging Alzheimer&apos s Disease Animals Architecture Bioinformatics Biological Assay Caenorhabditis elegans Caenorhabditis elegans Proteins Cellular Stress Cellular Stress Response Comparative Biology Complex Data Discrimination Disease Environment Environmental Health Etiology Exposure to Feedback Fluorescence Foundations Gene Expression Gene Targeting Genes Genetic Genetic Epistasis Genome Genomics Goals Green Fluorescent Proteins Health Heat Stress Disorders Heating Homeostasis Human Huntington Disease Knowledge Life Measurement Mediating Microarray Analysis Modeling Molecular Mutation National Institute of Environmental Health Sciences Nematoda Organism Oxidative Stress Parkinson Disease Pathway interactions Physiological Physiological Processes Play Poison Predisposition Principal Investigator Process Protein-Protein Interaction Map Proteins RNA Interference Regulation Regulatory Pathway Reporter Research Personnel Resistance Role Screening procedure Set protein Signal Pathway Signal Transduction Soil Stress System Testing Toxin Transgenic Animals Transgenic Organisms Work base biological adaptation to stress comparative computerized tools data integration disease registry environmental stressor functional genomics genome sequencing genome-wide human disease in vivo mutant overexpression polyglutamine prevent programs promoter protein aggregation protein folding protein function protein misfolding repair enzyme repaired response stressor transcription factor

项目摘要

DESCRIPTION (provided by applicant): Many human diseases, such as Huntington's, Parkinson's, and Alzheimer's, and normal physiological states, such as aging, are strongly influenced by exposure to environmental stress. However, little is known about the mechanisms by which environmental stressors are detected and discriminated from one another in animals. The long term objective of this work is to utilize functional genomic approaches in the nematode C. elegans to define fundamental environmental stress sensing and signaling mechanisms. Such studies will more precisely define the role of the environment in human health and disease and help to define the modes of action of numerous toxic substances. The specific hypothesis to be tested in this proposal is that compartment specific protein damage constitutes a feedback inhibition mechanism for discrimination between different forms of environmental stress. This hypothesis is based on the observations that 1) knockdown of protein homeostasis (PH) genes that normally prevent the accumulation of some forms of protein damage activates osmosensitive gene expression 2) Knockdown of PH genes that activate osmosensitive gene expression accelerates protein aggregation 3) Protein damage induced by heat and oxidative stress does not activate osmosensitive gene expression. The specific aims of this proposal are: 1) Define the PH genes that are transcriptionally regulated by environmental stressors. We will examine the spatial and temporal, and functional involvement of PH genes in response to environmental stress using i) comparative whole genome microarray analysis of heat, oxidative, and osmotically induced gene expression in wild type and transcription factor mutant animals ii) in vivo localization of GFP tagged PH genes during exposure to environmental stress iii) stress survival assays in transgenic animals overexpressing PH targets. 2) Test the role of protein misfolding as an activator of stress induced gene expression. Protein damage will be genetically induced using RNAi knockdown of specific PH complexes. The effects of each gene knockdown on the activation of heat, oxidative, and osmotic stress signaling pathways will be quantified by i) in vivo high throughput fluorescence measurements of animals expressing stress-specific GFP reporters ii) epistasis analysis using stress sensing transcription factor mutants iii) whole animal survival assays in PH knockdown animals.

描述（由申请人提供）：许多人类疾病，如亨廷顿病、帕金森病和阿尔茨海默病，以及正常的生理状态，如衰老，都受到环境压力的强烈影响。然而，人们对动物中检测和区分环境压力源的机制知之甚少。这项工作的长期目标是利用线虫的功能基因组方法。秀丽隐杆线虫来定义基本的环境应激感知和信号传导机制。此类研究将更准确地确定环境在人类健康和疾病中的作用，并有助于确定多种有毒物质的作用方式。该提案要测试的具体假设是，区室特异性蛋白质损伤构成了区分不同形式的环境应激的反馈抑制机制。该假设基于以下观察结果：1) 通常防止某些形式的蛋白质损伤积累的蛋白质稳态 (PH) 基因的敲低会激活渗透敏感基因表达 2) 激活渗透敏感基因表达的 PH 基因敲低会加速蛋白质聚集 3) 蛋白质热和氧化应激引起的损伤不会激活渗透敏感基因的表达。该提案的具体目标是：1）定义受环境应激源转录调节的 PH 基因。我们将使用 i) 对野生型和转录因子突变动物中热、氧化和渗透诱导的基因表达进行比较全基因组微阵列分析 ii) 体内定位来检查 PH 基因响应环境应激的空间和时间以及功能参与暴露于环境应激期间 GFP 标记的 PH 基因的变化 iii) 过度表达 PH 目标的转基因动物中的应激生存测定。 2) 测试蛋白质错误折叠作为应激诱导基因表达激活剂的作用。蛋白质损伤将通过特定 PH 复合物的 RNAi 敲低来基因诱导。每个基因敲低对热、氧化和渗透应激信号通路激活的影响将通过 i) 对表达应激特异性 GFP 报告基因的动物进行体内高通量荧光测量 ii) 使用应激传感转录因子突变体进行上位分析 iii 进行量化) PH 敲低动物的整体动物存活测定。