Molecular mechanisms of translational regulation in aging

衰老转化调控的分子机制

• 批准号: 10552685
• 负责人: Vyacheslav M Labunskyy
• 金额: $ 55.46万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552685
• 关键词: Address; Affect; Age; Aging; Amino Acids; Applications Grants; Bayesian Modeling; Binding; Binding Proteins; Binding Sites; Biological Assay; Biological Models; Candidate Disease Gene; Cardiovascular Diseases; Cells; Chimeric Proteins; Cytoprotection; Data; Development; Dietary Intervention; Disease; Elements; Fluorescence Microscopy; Functional disorder; Gene Deletion; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Epistasis; Genetic Transcription; Goals; Invertebrates; Knowledge; Link; Longevity; Malignant Neoplasms; Mammals; Maps; Mediating; Messenger RNA; Modeling; Molecular; Monitor; Mothers; Mutation; Nerve Degeneration; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Organism; Other Genetics; Pathway interactions; Pattern; Play; Post-Transcriptional Regulation; Process; Protein Biosynthesis; Proteins; RNA; RNA-Binding Proteins; Regulation; Regulatory Element; Reporter; Reproducibility of Results; Reverse engineering; Role; Signal Pathway; Starvation; System; Systems Biology; Testing; Time; Transcriptional Regulation; Translating; Translational Regulation; Translational Repression; Translations; Yeasts; aged; bioinformatics tool; cell age; data integration; dietary restriction; differential expression; experimental study; fly; gene therapy; genetic regulatory protein; genome wide screen; genome-wide; healthspan; human old age (65+); improved; in vivo; insight; intervention effect; mRNA Translation; microfluidic technology; mutant; next generation sequencing; posttranscriptional; programs; response; ribosome profiling; transcription factor; transcriptome sequencing

Molecular mechanisms of translational regulation in aging Genome-wide microarray and RNA sequencing studies have revealed changes in the expression of hundreds of genes during aging in diverse organisms. Transcriptional regulation plays an important role in the control of gene expression during aging; however, translation efficiency likely plays an equally important role in determining protein abundance, but has been understudied in this context. Here we propose to study translational changes that are associated with increased longevity induced by dietary and genetic interventions and examine the mech- anisms of post-transcriptional gene regulation in aging using yeast as a model system. We will test the hypoth- esis that, in response to dietary restriction (DR) and genetic alterations that extend lifespan, mRNA-binding pro- teins (RBPs) coordinately regulate diverse cytoprotective genes by affecting their translation efficiency. To iden- tify RBPs involved in regulation of these processes, we will apply RNA-Seq and ribosome profiling combined with next-generation sequencing and characterize transcriptional and translational changes in replicatively aged yeast cells in response to DR and in a panel of long-lived gene deletion mutants identified in genome-wide screens. Using the Bayesian network modeling approach, we will integrate transcriptional and translational pro- filing data with information about transcription factor (TF) binding sites and sequence elements recognized by RBPs and build a regulatory interaction network. We will then characterize RBPs and directly identify their mRNA-binding targets. These data will allow us to uncover specific mechanisms and identify cis-regulatory ele- ments that are responsible for the translational signatures associated with increased longevity. We will also utilize fluorescence microscopy and cutting-edge microfluidic technologies in order to monitor how the abun- dance of RBPs changes with age at the single-cell level. Finally, we will use reverse engineering approach to test if the regulators identified using our systems approach play a causative role in mediating the lifespan exten- sion through genetic epistasis experiments. Successful completion of this study will add valuable insight into fundamental principles of translational regulation, and may provide a better understanding of the molecular mechanisms of aging and lifespan control.

衰老过程中翻译调控的分子机制 全基因组微阵列和RNA测序研究揭示了数百个基因表达的变化 在不同的生物体中衰老过程中的基因。转录调控在控制病毒感染中起着重要作用。 基因在衰老过程中的表达；然而，翻译效率在决定 蛋白质丰度，但在这一背景下研究不足。在这里，我们建议研究翻译变化 与饮食和遗传干预导致的寿命延长有关，并检查了机制- 以酵母为模型系统研究衰老过程中转录后基因调控的失调性。我们将检验这一假设- 据认为，作为对饮食限制(DR)和延长寿命的基因改变的回应，mRNA结合蛋白- 蛋白质组(RBP)通过影响不同的细胞保护基因的翻译效率对其进行协调调节。为了认识到- 由于限制性商业惯例参与了这些过程的调节，我们将联合应用RNA-Seq和核糖体图谱 用下一代测序和表征复制老化的转录和翻译变化 酵母细胞对DR的反应以及在全基因组中发现的一组长寿命基因缺失突变 屏幕。使用贝叶斯网络建模方法，我们将整合转录和翻译过程。 使用关于转录因子(TF)结合位点和识别的序列元件的信息来归档数据 限制性商业惯例，并建立监管互动网络。然后，我们将描述RBP的特征并直接确定其 信使核糖核酸结合靶。这些数据将使我们能够揭示特定的机制，并确定顺式调控元件。 负责与延长寿命相关的翻译签名的片段。我们还将 利用荧光显微镜和尖端微流控技术来监测ABUN- 在单细胞水平上，限制性商业惯例的舞蹈随年龄变化。最后，我们将使用逆向工程方法来 测试使用我们的系统方法确定的监管机构是否在调解寿命延长- Sion通过遗传上位性实验。这项研究的成功完成将增加对 翻译调控的基本原理，并可能提供对分子更好的理解 衰老和寿命控制的机制。