Dissecting the genetic and molecular networks underlying longevity and aging

剖析长寿和衰老背后的遗传和分子网络

基本信息

批准号：
9145438
负责人：
Harmen J Bussemaker
金额：
$ 55.65万
依托单位：
PRINCETON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9145438
关键词：
Acetylesterase Adult Affect Affinity Chromatography Age Aging Amino Acid Sequence Animal Model Animals Binding Binding Proteins Biochemical Biological Assay Caenorhabditis elegans Cell Nucleus Cells Co-Immunoprecipitations Complement Coupled Cytoplasm DNA Binding DNA Sequence Alteration Data Development Disease Eating Elements Employee Strikes Enzymes Equilibrium Exhibits Gene Expression Profiling Gene Targeting Genes Genetic Genetic Models Genotype Goals Growth Growth and Development function Health Homologous Gene Human Insulin Insulin-Like Growth Factor I Intestines Laboratories Life Life Cycle Stages Longevity MAP Kinase Gene MHC Class I Genes MHC Class II Genes Mammalian Cell Mammals Mass Spectrum Analysis Methods Microfluidics Microscopy Modeling Molecular Molecular Genetics Molecular Profiling Nematoda Nuclear Nuclear Translocation Nutrient Organism Pathway interactions Pattern Phosphorylation Site Phosphotransferases Play Post-Translational Protein Processing Process Proteins Quantitative Trait Loci RNA Interference Recovery Regulation Regulatory Pathway Reporter Role Signal Pathway Signal Transduction Stress System Testing Time Tissues Transcription Coactivator Ursidae Family Variant Work age effect base biological adaptation to stress genome-wide high throughput screening human disease in vivo innovation insight luminescence mutant normal aging novel response small molecule transcription factor

项目摘要

DESCRIPTION (provided by applicant): Reduced insulin/IGF-1-like signaling (IIS) extends C. elegans lifespan by up-regulating stress response (Class I) and down-regulating development (Class II) genes through a mechanism that depends on the conserved transcription factor DAF-16/FOXO. By integrating a genomewide analysis of gene expression responsiveness to DAF-16 with genomewide in vivo binding data for a compendium of transcription factors, we discovered that the transcriptional activator PQM-1 directly controls Class II genes by binding to the DAF-16 associated element (DAE). DAF-16 directly regulates Class I genes only, through the DAF-16 binding element (DBE). Loss of PQM-1 suppresses daf-2 and eat-2 longevity as well as thermotolerance, and further slows development. The nuclear presence of PQM-1 and DAF- 16 is controlled by IIS in opposite ways, and, surprisingly, was found to be mutually exclusive. We also observe progressive loss of nuclear PQM-1 with age, explaining declining expression of PQM-1 targets. Together, our data suggest an elegant mechanism for switching between stress response and development. The overall goal of this project is to elucidate the mechanisms underlying the observed antagonistic relationship between PQM-1 and DAF-16. We will employ high-throughput screens based on reporter assays and microfluidics microscopy to identify genetic and small-molecule regulators of PQM-1 translocation. Additionally, we will use mass spectrometry to identify PQM-1's post-translational modifications and protein interactors, and determine how these affect nuclear translocation. We will also identify factors responsible for the nuclear exit of DAF-16 and PQM-1 with age. Finally, we will identify PQM-1 homologs in mammalian cells that exhibit a similar antagonism with FOXOs. Together, our results and insights will provide a framework for understanding how PQM-1 and DAF-16 and its mammalian counterparts allow cells to strike a balance between development and stress response.

描述（由申请人提供）：减少胰岛素/IGF-1样信号传导（IIS）通过上调应激反应（I类）和下调发育（II类）基因来延长秀丽隐杆线虫的寿命，该机制取决于保守转录因子 DAF-16/FOXO 通过将 DAF-16 基因表达反应性的全基因组分析与转录因子纲要的全基因组体内结合数据相结合，我们发现转录激活剂 PQM-1 通过与 DAF-16 相关元件 (DAE) 结合直接控制 II 类基因，DAF-16 仅通过 DAF-16 结合元件 (DBE) 抑制 PQM-1 的丢失。 daf-2 和 eat-2 的寿命以及耐热性，并进一步减慢发育 PQM-1 和 DAF-16 的核存在由 IIS 以相反的方式控制，并且，令人惊讶的是，我们还观察到核 PQM-1 随着年龄的增长而逐渐丧失，这解释了 PQM-1 靶点表达的下降，我们的数据表明了一种在应激反应和发育之间切换的优雅机制。该项目的目的是阐明 PQM-1 和 DAF-16 之间观察到的拮抗关系的机制，我们将采用基于报告分析和微流体显微镜的高通量筛选来识别 PQM-1 和 DAF-16 的遗传和小分子调节因子。此外，我们将使用质谱法来识别 PQM-1 的翻译后修饰和蛋白质相互作用因子，并确定它们如何影响核转位。我们还将确定导致 DAF-16 和 PQM- 出核的因素。最后，我们将鉴定哺乳动物细胞中与 FOXO 表现出类似拮抗作用的 PQM-1 同源物，我们的结果和见解将为理解如何发挥作用提供一个框架。 PQM-1和DAF-16及其哺乳动物成员使细胞能够在发育和应激反应之间取得平衡。