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

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.

摘要 非整倍性，即细胞携带不正确数量的染色体的状态，是人类染色体的主要问题。 健康非整倍体在哺乳动物发育过程中是有毒的，是导致妊娠丢失的主要原因。下来 21三体综合征（DS）是少数几个可行的非整倍体综合征之一，但受影响的个人有 包括过早衰老在内的终身问题。尽管进行了大量的研究，但非整倍体毒性的原因是 仍然不完全理解，对理解DS提出了挑战。相比之下，非整倍体非常 这在人类癌症中很常见，大多数肿瘤都能耐受，甚至可能受益于额外的染色体。是 不清楚癌细胞如何克服非整倍体的压力，因为我们不完全了解如何 非整倍性首先影响细胞。该提案将利用一个非常强大和独特的系统， 在一个重要的模式系统中研究非整倍体的后果，即芽殖酵母的野生菌株 酿酒酵母酵母是一个强大的模型，解剖细胞生物学，因为许多 机制和防御策略在人类中是保守的。我们最近有了一个令人兴奋的发现， 健康酵母菌株中的染色体复制破坏了营养反应和静止， 保守的细胞程序，对生长控制、细胞维持和更新很重要。多种菌株 遗传背景和携带不同的染色体扩增显示共同的表型，包括 营养耗尽后不完全的细胞周期停滞，代谢畸变，静止诱导的缺陷， 沉默，并最终减少按时间顺序的寿命。这是值得注意的，因为类似的缺陷 静止期的标志物在DS和许多癌症中都可以看到-如果静止期的破坏是保守的， 对于非整倍体的反应，它可能对未来的研究产生变革性的影响。这份协议将阐明如何 非整倍性破坏了重要的真核模型系统中的静止。目标1将使用动态 转录组学和单细胞显微镜来表征缺陷的时间顺序，测试几个初始的 假设，并牵连上游监管机构。它还将区分普通与染色体特异性 方面的影响. AIM 2将使用条形码质粒过表达文库来鉴定与 非整倍体缺陷沿着向静止的进展。整合目标1和目标2的结果将定义时间 非整倍体酵母菌株中与静止期有关的基因和过程缺陷图。它还将指出 染色体复制直接引起的上游缺陷，其进一步的研究将扩大我们的研究范围。 Aim 3将利用基因组学、蛋白质组学、单细胞和单分子分析， 定义并表征了“Ssd 1 Q颗粒”，一种含有RNA结合蛋白的相分离颗粒 ssd 1，我们以前表明是健康酵母的非整倍体耐受性的基础。由于许多 酵母中的机制在包括人类在内的高等生物中是保守的，这个项目将扩大我们的基础 对非整倍体的理解，并对多种人类疾病产生深远的影响。