Functional Genomics of Stress Defense in Yeast

酵母应激防御的功能基因组学

• 批准号: 7614185
• 负责人: AUDREY P GASCH
• 金额: $ 31.26万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7614185
• 关键词: Adverse effects; Animal Model; Area; Biochemical; Biochemical Genetics; Biochemistry; Biology; Cell Survival; Cells; Cellular Stress; Complex; Computational Biology; Data Set; Defense Mechanisms; Dependence; Disease; Dissection; Dose; Eukaryota; Fostering; Foundations; Fungal Genome; Gene Expression; Generations; Genes; Genetic; Genome; Genomics; Goals; Growth; Health; Homeostasis; Human; Malignant Neoplasms; Maps; Mediating; Medicine; Memory; Modeling; Molecular Profiling; Monitor; Mutation; Myocardial Infarction; Operative Surgical Procedures; Organism; Pathway Analysis; Phase; Phenotype; Play; Process; Proteomics; Regulation; Resistance; Role; Screening procedure; Signal Transduction; Stress; Stroke; System; Techniques; Toxin; Trauma; Untranslated Regions; Yeasts; acute stress; base; biological adaptation to stress; chemotherapy; deletion library; disorder prevention; functional genomics; gene function; high throughput screening; memory acquisition; mutant; novel; prevent; promoter; research study; response; success; transcription factor

DESCRIPTION (provided by applicant): All organisms must protect their internal system from cellular stress. Whether stress arises from external toxins or mutation and disease, cells must sensitively monitor stress signals and mount the appropriate responses to maintain internal homeostasis. Despite the importance of stress defense, much remains unknown about the mechanisms eukaryotes use to survive stressful situations. Functional genomics has uncovered functions for many genes in various genomes, largely by characterizing gene function under standard conditions. However, a substantial fraction of genes remains uncharacterized, and many of these are likely to be involved in stress defense and thus have not been uncovered through traditional studies. This proposal will use high-throughput functional genomics, genomic expression analysis, computational biology, and techniques in genetics and biochemistry to identify and characterize genes involved in stress defense in yeast. Aim 1 will exploit two new phenotypes related to stress defense to uncover novel genes involved in eukaryotic stress survival. The first is a phenomenon known as `acquired stress resistance', in which cells exposed to a small dose of one stress become resistant to an otherwise lethal dose of a different stress. The second is a phenomenon in which cells retain a `memory' of stress resistance that persists for many generations after mild-stress treatment, even after the mild stress has been removed. We will use these phenotypes in high-throughput selections to identify yeast deletion mutants that cannot acquire or retain resistance to severe stress after mild-stress treatment. Identified genes, as well as known regulators of acquired stress resistance, will be characterized to define their precise roles in these phenomena. Cells respond to stress with a multi-facetted response. This response, including reorganization of genomic expression, is coordinated by a complex signaling network that responds to stress. Aim 2 will elucidate the intricate stress-activated signaling network in yeast that orchestrates genomic expression responses to stress. Regulators of stress-dependent genes will be identified by screening the yeast-deletion library for mutants unable to induce expression upon stress treatments. Identified regulators and various known network components will be organized into a putative signaling network, using numerous computational approaches. This network will be subsequently dissected and refined based on genomic, genetic, and biochemical studies. These experiments will help to elucidate the complex stress-activated signaling network in yeast, which serves as an excellent model for such networks in humans and other organisms, while developing computational approaches that are likely to advance this area of biology. As many of these responses are conserved in humans, these results will foster stress minimization and disease prevention in human medicine. Project Relevance: Many stress-defense mechanisms used by yeast are conserved in humans, and therefore the results of this proposal will provide a strong foundation for understanding, and eventually modulating, stress resistance for human health. These results will have broad application, from minimizing debilitating side effects of chemotherapy, to reducing trauma inflicted by invasive surgery, heart attacks and strokes, to preventing cancer. Furthermore, understanding how yeast sense and respond to stress is an excellent model for how human cells respond to analogous cellular stresses.

描述（由申请人提供）：所有生物体必须保护其内部系统免受细胞应激。无论压力来自外部毒素还是突变和疾病，细胞都必须敏感地监测压力信号，并做出适当的反应以维持内部稳态。尽管压力防御的重要性，但真核生物在压力环境中生存的机制仍然未知。功能基因组学主要通过在标准条件下表征基因功能来揭示各种基因组中许多基因的功能。然而，相当一部分基因仍然没有被鉴定，其中许多基因可能与压力防御有关，因此尚未通过传统研究发现。该提案将使用高通量功能基因组学，基因组表达分析，计算生物学以及遗传学和生物化学技术来识别和表征酵母中参与应激防御的基因。目的1将利用两种新的与应激防御相关的表型来揭示真核生物应激生存相关的新基因。第一种是一种称为“获得性应激抗性”的现象，即细胞受到小剂量的一种应激后，对另一种应激的致死剂量产生了抗性。第二种现象是，细胞在轻度压力处理后，甚至在轻度压力消除后，仍保留着对压力抵抗的“记忆”，这种记忆持续了许多代。我们将使用这些表型在高通量选择，以确定酵母缺失突变体，不能获得或保持抵抗严重的压力后，温和的压力处理。已确定的基因，以及已知的调节获得性应激抗性，将其特点，以确定其在这些现象中的确切作用。细胞对压力的反应是多方面的。这种反应，包括基因组表达的重组，是由一个复杂的信号网络，响应压力协调。目的2将阐明酵母中复杂的应激激活信号网络，协调基因组表达对应激的反应。将通过筛选酵母缺失文库中在胁迫处理后不能诱导表达的突变体来鉴定胁迫依赖性基因的调节子。确定的监管机构和各种已知的网络组件将被组织成一个假定的信号网络，使用多种计算方法。随后将根据基因组、遗传和生物化学研究对该网络进行解剖和细化。这些实验将有助于阐明酵母中复杂的应激激活信号网络，这是人类和其他生物体中此类网络的一个很好的模型，同时开发可能推进这一生物学领域的计算方法。由于许多这些反应在人类中是保守的，这些结果将促进人类医学中的压力最小化和疾病预防。 项目相关性：酵母所使用的许多应激防御机制在人类中是保守的，因此这项提议的结果将为理解并最终调节人类健康的应激抵抗力提供坚实的基础。这些结果将有广泛的应用，从最大限度地减少化疗的副作用，到减少侵入性手术，心脏病发作和中风造成的创伤，到预防癌症。此外，了解酵母如何感知和应对压力是人类细胞如何应对类似细胞压力的绝佳模型。