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.

衰老中翻译调节的分子机制 全基因组微阵列和RNA测序研究揭示了数百种表达的变化 不同生物体衰老过程中的基因。转录调节在控制中起着重要作用 衰老期间的基因表达；但是，翻译效率可能在确定中起着同样重要的作用 蛋白质丰度，但在这种情况下已经被研究了。在这里，我们建议研究翻译变化 与饮食和遗传干预引起的寿命增加有关的，并检查机甲 使用酵母作为模型系统衰老中转录后基因调节的疾病。我们将测试假设 ESI，应响应饮食限制（DR）和延长寿命的遗传改变，mRNA结合促进 Teins（RBPS）通过影响其翻译效率来协调调节多样化的细胞保护基因。到 TIFY参与这些过程调节的RBP，我们将应用RNA-Seq和核糖体分析合并 下一代测序并表征复制年龄的转录和翻译变化 酵母细胞响应DR和在全基因组中鉴定出的一组长寿命缺失突变体中 屏幕。使用贝叶斯网络建模方法，我们将整合转录和翻译促销 使用有关转录因子（TF）结合位点的信息和序列元素的归档数据 RBP并建立一个监管交互网络。然后，我们将表征RBP并直接识别他们的 mRNA结合靶标。这些数据将使我们能够发现特定的机制并确定顺式调节性ELE- 负责与寿命增加相关的翻译特征的损失。我们也会 利用荧光显微镜和尖端的微流体技术，以监测 RBP的舞蹈随着单细胞水平的年龄而变化。最后，我们将使用逆向工程方法进行 测试使用我们的系统方法确定的调节器是否在介导寿命延长的情况下起着致命作用 通过遗传性上述实验进行的。这项研究的成功完成将增加对 翻译调节的基本原理，可以更好地理解分子 衰老和寿命控制的机制。