Dynamics of Ribosome Biogenesis in Yeast

酵母中核糖体生物发生的动力学

• 批准号: 8373500
• 负责人: CURT WITTENBERG
• 金额: $ 42.72万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8373500
• 关键词: Address; Ammonium; Binding; Biogenesis; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Nucleus; Cell Proliferation; Cell division; Cells; Characteristics; Cytoplasm; Data; Defect; Escherichia coli; Eukaryota; Event; Genetic; Goals; Growth; Isotopes; Label; Maps; Mass Spectrum Analysis; Mitotic Cell Cycle; Modeling; Monitor; Pathway interactions; Physiologic pulse; Population; Process; Protein Biosynthesis; Proteins; RNA Processing; Regulation; Relative (related person); Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Sucrose; Work; Yeasts; abstracting; cell growth; cell type; cofactor; genetic analysis; particle; programs; tool; yeast genetics

DESCRIPTION (provided by applicant): Project Summary/Abstract This proposal outlines a plan to study the dynamics of ribosome assembly in yeast, and the relationship between ribosome biogenesis and growth and cell cycle control. The synthesis of ribosomes in eukaryotes requires > 200 assembly factors to direct the ribosomal RNA processing and ordered assembly of >100 ribosomal proteins. While the framework for rRNA processing is well established, there is no coherent mechanism for the dynamics of biogenesis factor binding and release or association of the ribosomal proteins. Furthermore, the control of the ribosome biogenesis program is poorly understood in two key respects. First, the mechanism by which synthesis of the ribosomal proteins in the cytoplasm is coordinated with the dynamics for ribosome assembly in the nucleus is poorly understood. Second, although there is ample evidence for the association between defects in ribosome biogenesis and defects in cell growth and division, the mechanism by which ribosome biogenesis is monitored to regulate growth and progression through the cell cycle is unclear. In this proposal, we will apply quantitative analysis of ribosome biogenesis using mass spectrometry with yeast genetic analysis to address both of these key questions. This will be accomplished with three Specific Aims: 1) Implement Isotope Pulse-Quantitative Mass Spectrometry (IP-QMS) for monitoring ribosome biogenesis in yeast. 2) Elaborate the mechanism of ribosome biogenesis by defining the pathway(s) for assembly of ribosomal proteins. 3) To apply IP-QMS to analyze the roles of ribosome biogenesis factors and place them on the assembly pathway map developed in Aim 2 and to elucidate the pathways coordinating regulated ribosome biogenesis with growth and the cell cycle. The main goal of this proposal is to combine yeast genetics and QMS to interrogate the intermediates in ribosome biogenesis involving ribosomal proteins and biogenesis factors and to identify connections between those intermediates and the regulation of cell growth and the cell cycle. PUBLIC HEALTH RELEVANCE: Project Narrative This proposal outlines a plan to study the dynamics of ribosome assembly in yeast, and the relationship between ribosome biogenesis and growth and cell cycle control. Regulating protein synthesis capacity and production of ribosomes is essential for all cell types. Understanding abnormal regulation of ribosome biogenesis has implications for rapid cell growth in cancer, and for hematopoiesis in Diamond-Blackfan Anemia.

描述（由申请人提供）： 项目概要/摘要本提案概述了研究酵母中核糖体组装动力学的计划，以及核糖体生物发生与生长和细胞周期控制之间的关系。真核生物中核糖体的合成需要> 200个组装因子来指导核糖体RNA加工和>100个核糖体蛋白的有序组装。虽然rRNA加工的框架已经很好地建立，但生物合成因子结合和释放或核糖体蛋白结合的动力学没有连贯的机制。此外，核糖体生物发生程序的控制在两个关键方面知之甚少。首先，核糖体蛋白质在细胞质中的合成与核糖体在细胞核中组装的动力学协调的机制知之甚少。其次，虽然有充分的证据表明核糖体生物合成缺陷与细胞生长和分裂缺陷之间存在关联，但通过监测核糖体生物合成来调节细胞周期中的生长和进展的机制尚不清楚。在这个建议中，我们将应用定量分析的核糖体生物利用质谱与酵母遗传分析，以解决这两个关键问题。这将通过三个具体目标来实现：1）实施同位素脉冲定量质谱法（IP-QMS），用于监测酵母中的核糖体生物合成。2)通过定义核糖体蛋白组装的途径，阐述核糖体生物发生的机制。3)应用IP-QMS分析核糖体生物合成因子的作用，并将其置于Aim 2中开发的组装通路图上，阐明调控核糖体生物合成与生长和细胞周期的协调通路。该建议的主要目标是将联合收割机酵母遗传学和QMS相结合，以询问涉及核糖体蛋白和生物合成因子的核糖体生物合成中的中间体，并确定这些中间体与细胞生长和细胞周期调节之间的联系。 公共卫生关系： 该提案概述了研究酵母中核糖体组装动力学的计划，以及核糖体生物发生与生长和细胞周期控制之间的关系。调节蛋白质合成能力和核糖体的产生对所有细胞类型都是必不可少的。了解核糖体生物合成的异常调节对癌症中的快速细胞生长和Diamond-Blackfan贫血中的造血有意义。