Ribosomal RNA methylation regulation of longevity and stress resistance

核糖体RNA甲基化对长寿和抗压能力的调节

• 批准号: 10688324
• 负责人: Eric Lieberman Greer
• 金额: --
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10688324
• 关键词: Adenosine; Age; Aging; Animals; Binding; Biochemical; Biological; Caenorhabditis elegans; Cells; Complex; Coupled; Cues; DNA Methylation; Data; Enzymes; Eukaryota; Event; Genetic; Genetic Translation; Goals; Health; Heat-Shock Response; Heterogeneity; High Pressure Liquid Chromatography; In Vitro; Individual; Lead; Longevity; Maintenance; Mass Spectrum Analysis; Messenger RNA; Metabolism; Methods; Methylation; Methyltransferase; Modification; Molecular; Mus; Organism; Outcome; Pathway interactions; Play; Process; Protein Biosynthesis; Proteins; Proteome; RNA interference screen; RNA methylation; RNA, Ribosomal, 18S; Regulation; Resistance; Ribosomal DNA; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Signal Pathway; Specific qualifier value; Specificity; Stimulus; Stress; Structure; Testing; Tissues; Transcript; Transgenic Organisms; Translating; Translations; Work; biological adaptation to stress; environmental stressor; experimental study; gene product; healthy aging; in vivo; insight; mutant; polysome profiling; preservation; response; ribosome profiling; transcriptome; ultraviolet irradiation

Project Summary The goal of this project is to understand how ribosomal RNA methylation can regulate translation of specific proteins to regulate aging. Disruption of the proteome is a hallmark of aging. Having the capacity to express the appropriate protein in response to environmental cues is an essential and evolutionarily conserved process. Therefore, preserving the proteome is critical for maintaining organismal health and healthy aging. However, how aging-responsive mRNAs are selectively translated is unknown. We recently identified that ribosomal RNA methylation could facilitate the translation of a specific subset of proteins. Additionally, we characterized how ribosome occupancy changes as an organism ages and determined that changes in ribosomal DNA methylation are sufficient to project the organismal age. Whether ribosomal RNA methylation plays a role in preserving proteome integrity as organisms age is still unclear. We have recently found that enzymes which regulate RNA methylation and RNA methylation itself are dysregulated during aging and in response to stress. We found that an N6-adenosine methyltransferase, METL-5, directly methylates adenosine 1717 on 18S ribosomal RNA in C. elegans. Methylation of adenosine 1717 enhances ribosomal binding and selective translation of specific mRNAs. Our preliminary data shows that metl-5 deficient animals grow normally under homeostatic conditions; however, metl-5 mutants are resistant to a variety of stresses, including heat shock and UV irradiation. Thus, methylation of a specific residue of 18S rRNA by METL-5 selectively enhances translation of specific transcripts to regulate stress resistance. However, whether rRNA methylation more broadly can regulate age and stress-responsive translation and whether this dysregulation drives the aging process is still unknown. By performing a directed RNAi screen, we identified additional rRNA methyltransferases that when deleted increase C. elegans lifespan. These preliminary findings suggest that ribosomal RNA methylation can facilitate selective translation of specific transcripts providing another layer of regulation of the stress response and aging. Capitalizing on this preliminary data we will use genetic, biochemical, and molecular approaches both in vitro and in vivo to dissect the role of rRNA methylation in regulating aging and stress resistance. Our underlying hypothesis is that rRNA methylation promotes ribosome heterogeneity in response to stress conditions and aging to facilitate the appropriate translation of stress resistance transcripts.

项目摘要 该项目的目的是了解核糖体RNA甲基化如何调节特定的翻译 蛋白质调节衰老。蛋白质组的破坏是衰老的标志。有能力表达 响应环境线索的适当蛋白质是必不可少的且进化保守的 过程。因此，保留蛋白质组对于维持有机体健康和健康衰老至关重要。 但是，如何选择性地翻译衰老响应性mRNA是未知的。我们最近确定 核糖体RNA甲基化可以促进蛋白质特定子集的翻译。另外，我们 表征了核糖体占用率如何随着生物体的年龄变化而确定的变化 核糖体DNA甲基化足以投射有机年龄。核糖体RNA甲基化是否 由于生物年龄仍然不清楚，因此在保持蛋白质组完整性方面起作用。我们最近发现 调节RNA甲基化和RNA甲基化本身的酶在衰老期间和在 对压力的反应。我们发现N6-腺苷甲基转移酶Metl-5直接甲基化腺苷 1717年在秀丽隐杆线虫中的18S核糖体RNA。 1717腺苷的甲基化增强了核糖体结合和 特定mRNA的选择性翻译。我们的初步数据表明，Metl-5不足动物生长 通常在稳态条件下；但是，Met-5突变体对各种应力有抵抗力， 包括热休克和紫外线照射。因此，通过metl-5对18S rRNA的特定残基的甲基化 有选择地增强特定转录本的翻译以调节应力抗性。但是，是否rRNA 甲基化更广泛地调节年龄和压力响应性翻译，以及这种失调是否失调 驱动衰老过程仍然未知。通过执行定向RNAi屏幕，我们确定了其他rRNA 当删除时，甲基转移酶会增加秀丽隐杆线虫的寿命。这些初步发现表明 核糖体RNA甲基化可以促进特定转录本的选择性翻译，从而提供另一层 调节压力反应和衰老。利用这些初步数据，我们将使用遗传， 生化和分子在体外和体内都可以剖析rRNA甲基化的作用 调节衰老和压力抗性。我们的基本假设是rRNA甲基化促进核糖体 响应压力条件和衰老的异质性，以促进应力的适当翻译 电阻转录本。