Understanding how aneuploidy disrupts quiescence in the model eukaryote Saccharomyces cerevisiae

了解非整倍体如何破坏模型真核生物酿酒酵母的静止状态

• 批准号: 10735074
• 负责人: AUDREY P GASCH
• 金额: $ 30.03万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10735074
• 关键词: Affect; Aneuploidy; Bar Codes; Biological Models; Buffers; Cell Cycle Arrest; Cell Cycle Regulation; Cell Maintenance; Cell division; Cells; Cellular biology; Chromosome Mapping; Chromosome Segregation; Chromosomes; Chronology; Complement; Cytoplasmic Granules; Defect; Development; Disease; Down Syndrome; Drug resistance; Edward' s syndrome; Eukaryota; Evolution; Expression Library; Fertility Disorders; Future; Genes; Genetic; Genomics; Glucose; Goals; Grant; Growth; Health; Human; Individual; Karyotype; Link; Longevity; Malignant Neoplasms; Maps; Meiosis; Metabolic; Methods; Microscopy; Mitochondria; Mitosis; Modeling; Molecular; Nutrient; Nutrient Depletion; Organism; Patau' s syndrome; Phase; Phenotype; Plasmids; Play; Pregnancy loss; Premature aging syndrome; Process; Proteins; Proteome; Proteomics; RNA; RNA-Binding Proteins; Respiration; Role; Saccharomyces cerevisiae; Saccharomycetales; Stress; Syndrome; System; Testing; Toxic effect; Work; Yeasts; cancer cell; daughter cell; developmental disease; exhaustion; experience; gene complementation; human disease; human pathogen; live cell imaging; overexpression; programs; promoter; response; single molecule; stem cell function; tissue regeneration; transcriptome; transcriptome sequencing; transcriptomics; tumor; Affect; Aneuploidy; Bar Codes; Biological Models; Buffers; Cell Cycle Arrest; Cell Cycle Regulation; Cell Maintenance; Cell division; Cells; Cellular biology; Chromosome Mapping; Chromosome Segregation; Chromosomes; Chronology; Complement; Cytoplasmic Granules; Defect; Development; Disease; Down Syndrome; Drug resistance; Edward' s syndrome; Eukaryota; Evolution; Expression Library; Fertility Disorders; Future; Genes; Genetic; Genomics; Glucose; Goals; Grant; Growth; Health; Human; Individual; Karyotype; Link; Longevity; Malignant Neoplasms; Maps; Meiosis; Metabolic; Methods; Microscopy; Mitochondria; Mitosis; Modeling; Molecular; Nutrient; Nutrient Depletion; Organism; Patau' s syndrome; Phase; Phenotype; Plasmids; Play; Pregnancy loss; Premature aging syndrome; Process; Proteins; Proteome; Proteomics; RNA; RNA-Binding Proteins; Respiration; Role; Saccharomyces cerevisiae; Saccharomycetales; Stress; Syndrome; System; Testing; Toxic effect; Work; Yeasts; cancer cell; daughter cell; developmental disease; exhaustion; experience; gene complementation; human disease; human pathogen; live cell imaging; overexpression; programs; promoter; response; single molecule; stem cell function; tissue regeneration; transcriptome; transcriptome sequencing; transcriptomics; tumor

ABSTRACT Aneuploidy, the state in which cells carry an incorrect number of chromosomes, is a major problem for human health. Aneuploidy is toxic during mammalian development and a leading cause of pregnancy loss. Down syndrome (DS) due to trisomy 21 is one of the few viable aneuploid syndromes, but affected individuals have life-long problems including premature aging. Despite intense study, the reasons for aneuploidy toxicity are still incompletely understood, presenting challenges for understanding DS. In contrast, aneuploidy is very common in human cancers, where most tumors tolerate and may even benefit from extra chromosomes. It is unclear how cancer cells overcome the stress of aneuploidy, because we don’t fully understand how aneuploidy affects cells in the first place. This proposal will utilize an extremely powerful and unique system to study the consequence of aneuploidy in an important model system, wild strains of budding yeast Saccharomyces cerevisiae. Yeast is a powerful model for dissecting cellular biology, because many of the mechanisms and defense strategies are conserved in humans. We recently made an exciting discovery that chromosome duplication in healthy yeast strains disrupts nutrient responses and quiescence, a conserved cellular program important for growth control and cell maintenance and renewal. Strains of multiple genetic background and carrying different chromosome amplifications display shared phenotypes, including incomplete cell-cycle arrest upon nutrient depletion, metabolic aberrations, defects in quiescence-induced silencing, and ultimately reduced chronological life span. This is remarkable, because defects in similar markers of quiescence are seen in both DS and many cancers – if disruption of quiescence is a conserved response to aneuploidy, it could have transformative impacts for future studies. This grant will elucidate how aneuploidy disrupts quiescence in an important eukaryotic model system. Aim 1 will use dynamic transcriptomics and single-cell microscopy to characterize the temporal order of defects, test several initial hypotheses, and implicate upstream regulators. It will also distinguish common versus chromosome-specific effects. Aim 2 will use a barcoded plasmid over-expression library to identify genes that complement aneuploid defects along the progression to quiescence. Integrating Aim 1 and 2 results will define a temporal map of genes and processes defective in aneuploid yeast strains and involved in quiescence. It will also point to the upstream defect(s) directly caused by chromosome duplication, whose further study will expand our understanding of aneuplodiy Aim 3 will use genomic, proteomic, single-cell and single-molecule analysis to define and characterize the “Ssd1 Q granule”, a phase separated granule containing the RNA-binding protein Ssd1, which we previously showed is fundamental for aneuploidy tolerance in healthy yeast. Since many mechanisms in yeast are conserved in higher organisms including humans, this project will expand our basic understand of aneuploidy and have far-reaching impact relevant for multiple human diseases.

抽象的 非整倍性是细胞携带不正确数量染色体的状态，是人类的主要问题 健康。非整倍性在哺乳动物发育过程中是有毒的，也是妊娠丧失的主要原因。向下 综合征（DS）由于21造成 终身问题，包括过早衰老。尽管进行了深入研究，但非整倍性毒性的原因是 仍然不完全理解，给理解DS带来了挑战。相反，非整倍性非常 在人类癌症中常见，大多数肿瘤耐受性，甚至可能受益于额外的染色体。这是 尚不清楚癌细胞如何克服非整倍性的压力，因为我们不完全了解 非整倍性首先会影响细胞。该建议将利用一个极其强大且独特的系统来 在重要的模型系统中研究非整倍性的后果 酿酒酵母。酵母是剖析细胞生物学的强大模型，因为许多 机制和防御策略在人类中是保守的。我们最近做出了一个令人兴奋的发现 健康酵母菌菌株中的染色体重复破坏营养反应和静止 保守的细胞计划对于生长控制和细胞维持和更新很重要。多重菌株 遗传背景和携带不同的染色体扩增显示共享表型，包括 营养部署，代谢像差，静止诱导的缺陷后，细胞周期停滞不完全 沉默，最终降低了年代寿命。这是非常了不起的，因为相似的缺陷 在DS和许多癌症中都可以看到静止的标记 - 如果静止的破坏是保守的 对非整倍性的反应，它可能会对未来的研究产生变革性的影响。这笔赠款将阐明如何 非整倍性破坏了重要的真核模型系统中的静止。 AIM 1将使用动态 转录组学和单细胞显微镜以表征缺陷的临时顺序，测试几个初始 假设和暗示上游调节器。它还将区分常见和特定于染色体的公共 效果。 AIM 2将使用条形码的质粒过表达库来识别完成的基因 沿静止的苯并酶缺陷。集成AIM 1和2的结果将定义临时性 基因和过程的图图在非整倍体酵母菌菌株中有缺陷，并参与静止。它也会指出 直接由染色体重复引起的上游缺陷，其进一步的研究将扩大我们 对Aneuplodiy AIM 3的了解将使用基因组，蛋白质组学，单细胞和单分子分析 定义并表征“ SSD1 Q颗粒”，这是一种含有RNA结合蛋白的相分离的颗粒 我们先前显示的SSD1对于健康酵母中的非整倍性耐受性至关重要。自从很多 酵母中的机制在包括人类在内的较高生物体中是保守的，该项目将扩大我们的基本 了解非整倍性，并具有与多种人类疾病有关的深远影响。