Dynamics in Eukaryotic Dormancy: Gene Expression and Aging

真核休眠的动态：基因表达和衰老

• 批准号: 10670978
• 负责人: Hyun Youk
• 金额: $ 41.88万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10670978
• 关键词: Acceleration; Age; Aging; Basic Science; Biological Models; Biology; Cells; Deceleration; Face; Fungal Spores; Gene Deletion; Gene Expression; Gene Expression Regulation; Gene Library; Genes; Genetic Transcription; Goals; Health; Histoplasmosis; Human; Human body; Knowledge; Laboratories; Life; Longevity; Measurement; Meningitis; Messenger RNA; Methods; Microbe; Microscope; Mission; Nature; Nutrient; Organism; Pharmaceutical Preparations; Plants; Process; Production; Proteins; RNA; Reproduction spores; Research; Ribosomal Proteins; Role; Saccharomyces cerevisiae; Techniques; Translations; United States National Institutes of Health; Water; Work; Yeasts; antimicrobial drug; cancer cell; detection method; fungus; genome-wide; improved; insight; interest; mathematical model; microbial; novel therapeutics; transcriptome sequencing; virtual

PROJECT SUMMARY Dormancy is a state in which virtually all intracellular activities, such as gene expression, are thought to have (nearly) stopped. Many organisms and cells become dormant when they face dire conditions such as lack of nutrients. Despite its ubiquity, the state of dormancy remains poorly understood and underexplored. A major open question is which intracellular processes might still occur in dormancy, to what extent, and whether and how they are important for surviving dormancy. This question is relevant to dormancy of microbial spores, cancer cells, plant seeds, worms, cells in human body, and others. Microbial spores are particularly important because many microbes in nature often exist as dormant spores rather than as vegetative cells. Many fungal spores are of interest because they are infectious and are difficult to kill with existing drugs for unknown reasons. My laboratory's goal is to answer the critical question posed above for understanding dormancy. We use the dormant yeast (Saccharomyces cerevisiae) spores as a model system for studying eukaryotic dormancy. With dormant yeast spores, we focus on two fundamental aspects of life: (1) Dynamics and regulation of gene expression in dormancy (2) Dynamics and determinants of aging in dormancy My lab makes quantitative measurements at single-cell and genome-wide levels and combines them with mathematical models of gene regulations. We recently discovered that yeast spores express some genes while dormant (i.e., in water without any nutrients) and that, surprisingly, some of the expression levels can be as high as in vegetative yeasts. To extend this discovery, we adapted an RNA-Seq-based technique to detect all freshly made RNAs in dormant yeast spores. We discovered that dormant yeast spores transcribe ~65% of their genes, with ribosomal proteins being one of the most highly transcribed. With microscope-based techniques that detect mRNA and protein productions in the same single spore and mathematical models that screen various forms of gene regulation, we are now uncovering signs of globally (genome-wide) coordinated transcription and translation whose mechanisms we aim to elucidate in the next five years. Our ongoing work is also uncovering dormant yeast spores secreting molecules that help each other survive, extend lifespans, and regulate gene expression. We will elucidate the mechanisms of this "collective dormancy" and signs of aging in dormant yeast spores. A comprehensive library of gene-deleted yeast strains will help us determine how each gene accelerates or decelerates aging in dormant spores and its role in collective dormancy. Our work will advance the still- primitive understanding of eukaryotic, microbial dormancy by establishing foundational knowledge on gene regulation and aging in dormancy with quantitative approaches that have rarely been applied to these topics. More broadly, we expect that our work will provide conceptual insights into quiescent cells in general.

项目总结 休眠是一种几乎所有的细胞内活动，如基因表达，都被认为具有 (几乎)停了下来。许多生物体和细胞在面临可怕的条件时变得休眠，例如缺乏 营养素。尽管它无处不在，但人们对休眠状态仍然知之甚少，也没有得到充分的研究。一位少校 悬而未决的问题是，哪些细胞内过程可能仍然处于休眠状态，休眠程度如何，以及是否和 它们对于休眠的生存是多么的重要。这个问题与微生物孢子的休眠、癌症有关。 细胞、植物种子、蠕虫、人体细胞等。微生物孢子特别重要，因为 自然界中的许多微生物通常以休眠孢子而不是营养细胞的形式存在。许多真菌孢子是 引起人们的兴趣，因为它们具有传染性，很难用现有的药物杀死，原因不明。 我的实验室的目标是回答上面提出的理解休眠的关键问题。我们用 以休眠酵母(Saccharmyces Cerevisiae)孢子为模型系统研究真核生物休眠。 对于休眠的酵母孢子，我们关注生命的两个基本方面： (1)休眠过程中基因表达的动态与调控 (2)休眠中衰老的动力学及决定因素 我的实验室在单细胞和全基因组水平上进行定量测量，并将它们与 基因调控的数学模型。我们最近发现，酵母孢子表达一些基因，而 休眠(即在没有任何营养的水中)，令人惊讶的是，一些表达水平可以像 就像营养酵母一样。为了扩大这一发现，我们采用了一种基于RNA-Seq的技术来检测所有新鲜的 在休眠的酵母孢子中制造核糖核酸。我们发现，休眠的酵母孢子转录了~65%的基因， 其中核糖体蛋白是转录水平最高的蛋白质之一。利用基于显微镜的技术来检测 同一单孢子中的mRNA和蛋白质的产生以及筛选不同形式的 基因调控，我们现在正在发现全球(基因组范围)协调转录和 我们的目标是在未来五年阐明其机制。我们正在进行的工作也揭示了 休眠的酵母孢子分泌相互帮助生存、延长寿命和调节基因的分子 表情。我们将阐明这种“集体休眠”的机制和休眠酵母衰老的迹象。 孢子。一个全面的基因缺失酵母菌株文库将帮助我们确定每个基因是如何加速的 或减缓休眠孢子的衰老及其在集体休眠中的作用。我们的工作将推动仍然- 通过建立基因基础知识对真核生物、微生物休眠的初步认识 休眠中的调节和衰老与很少被应用于这些主题的量化方法。 更广泛地说，我们希望我们的工作将提供对静止细胞的概念性洞察。