Molecular mechanisms of translational regulation in aging

衰老转化调控的分子机制

• 批准号: 9902274
• 负责人: Vyacheslav M Labunskyy
• 金额: $ 56.04万
• 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 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9902274
• 关键词: Address; Affect; Age; Aging; Amino Acids; Applications Grants; Bayesian Modeling; Binding; Binding Sites; Biological Assay; Biological Models; Candidate Disease Gene; Cardiovascular Diseases; Cells; Chimeric Proteins; Data; Development; Dietary Intervention; Disease; Elements; Engineering; Fluorescence Microscopy; Functional disorder; Gene Deletion; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Epistasis; Genetic Transcription; Genetic Translation; 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; Role; Signal Pathway; Starvation; System; Systems Biology; Testing; Time; Transcriptional Regulation; Translating; Translational Regulation; Translations; Yeasts; aged; bioinformatics tool; cell age; dietary restriction; differential expression; experimental study; fly; gene therapy; genetic regulatory protein; genome wide screen; genome-wide; healthspan; improved; in vivo; insight; intervention effect; microfluidic technology; mutant; next generation sequencing; 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种基因表达的变化， 基因在不同生物体衰老过程中的变化。转录调控在调控细胞凋亡中起着重要作用。 衰老过程中的基因表达;然而，翻译效率可能在决定衰老过程中起着同样重要的作用。 蛋白质丰度，但在这方面研究不足。在这里，我们建议研究翻译的变化 与饮食和遗传干预引起的长寿有关，并检查了这种机制， 使用酵母作为模型系统的衰老中转录后基因调控的anisms。我们将检验这个假设- 研究表明，作为对饮食限制（DR）和延长寿命的遗传改变的反应， 蛋白质（RBP）通过影响其翻译效率来协调调节多种细胞保护基因。去确认- 为了验证参与这些过程调控的RBP，我们将结合RNA-Seq和核糖体分析， 与下一代测序和表征转录和翻译的变化， 酵母细胞对DR的反应，以及在全基因组中鉴定的一组长寿基因缺失突变体中， 卡位使用贝叶斯网络建模方法，我们将整合转录和翻译前， 用转录因子（TF）结合位点和序列元件的信息归档数据， 限制性商业惯例和建立监管互动网络。然后，我们将描述限制性商业惯例，并直接确定其 mRNA结合靶点。这些数据将使我们能够揭示特定的机制，并确定顺式调节元件。 负责与寿命增加相关的翻译签名的片段。我们还将 利用荧光显微镜和尖端的微流体技术，以监测如何abun- 在单细胞水平上，RBP的舞蹈随年龄而变化。最后，我们将使用逆向工程的方法， 测试使用我们的系统方法确定的调节因子是否在介导寿命延长中发挥因果作用， 锡永遗传上位性实验。成功完成这项研究将增加宝贵的见解， 翻译调控的基本原则，并可能提供更好的理解的分子 衰老和寿命控制的机制。