YEAST RIBOSOME BIOGENESIS

酵母核糖体生物发生

• 批准号: 7957743
• 负责人: JOHN L. WOOLFORD
• 金额: $ 0.7万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957743
• 关键词: ATP phosphohydrolase; Address; Binding; Biochemical; Biogenesis; Biological; Biology; Boxing; Cell Nucleolus; Computer Retrieval of Information on Scientific Projects Database; Cytoplasm; Disease; Eukaryota; Funding; Fungal Genome; Goals; Grant; Human; In Vitro; Institution; Malignant Neoplasms; Modification; Molecular Genetics; Neighborhoods; Nucleoplasm; Pathway interactions; Play; Process; Proteins; Proteomics; Recruitment Activity; Regulation; Research; Research Personnel; Resources; Ribosomal Proteins; Ribosomal RNA; Ribosomes; Role; Saccharomyces cerevisiae; Series; Source; Trans-Activators; United States National Institutes of Health; Yeasts; cell growth; method development; mutant; particle; premature; prevent; rRNA Precursor; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long term goal of this project is to understand how ribosomes are assembled in eukaryotes. We use the yeast Saccharomyces cerevisiae, to facilitate combined genetic, molecular biological, biochemical, and proteomic approaches. Ribosome assembly initiates in the nucleolus, where rRNA is transcribed, associates with ribosomal proteins, and undergoes modification and initial processing to begin to form mature ribosomal RNPs. Subsequent steps in maturation of preribosomal particles occur upon their release from the nucleolus to the nucleoplasm and upon their export to the cytoplasm. This assembly pathway requires a dynamic series of remodeling steps in which protein-protein, RNA-protein, and RNA-RNA interactions are established, disrupted and reconfigured. Dysregulation of this pathway in humans leads to many diseases related to alterations in cell growth or proliferation, including cancer. Screens for yeast mutants defective in ribosome biogenesis, and development of methods to purify ribosome assembly intermediates and identify their constituents, led to the identification of >170 trans-acting factors required for ribosome assembly. Central to understand the mechanisms of ribosome biogenesis will be to figure out the precise roles played by each of these assembly factors. Which of them contacts pre-rRNA? Which proteins interact with each other? Can one define assembly neighborhoods within the nascent rRNPs, as observed for prokaryotic ribosomes assembled in vitro? In what order do these factors associate with preribosomes, carry out their functions, then dissociate from the particles? By what means are assembly factors and ribosomal proteins recruited to pre-ribosomes, activated, then released from the pre-rRNPs? Experiments are proposed to address these questions. We are focusing on two particular consecutive steps in the maturation of precursors to mature 60S ribosomal subunits--maturation of the 66SA3 assembly intermediates, followed by the 66SB particles. To begin to establish paradigms for mechanisms of factor function, we will address the above questions for several factors required for these two steps in assembly. In addition, we will investigate in more detail the role of the DEAD-box protein Drs1. Does this putative ATPase enable maturation of 66SB particles, by triggering the release of negative regulators that bind to pre-rRNA and prevent its premature cleavage? The high degree of conservation of molecules involved in eukaryotic ribosome biogenesis promises that principles governing ribosome biogenesis discovered in yeast will provide blueprints to study more diverse modes of regulation of ribosome assembly in metazoans,including those disrupted in many diseases.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该项目的长期目标是了解真核生物如何组装核糖体。 我们使用酿酒酵母的酵母菌，促进遗传，分子生物学，生化和 蛋白质组学方法。 核糖体组装在核仁中启动，其中rRNA被转录，与核糖体蛋白相关，并经过修饰和初始加工，开始形成成熟的核糖体RNP。 释放后的后颗粒成熟的随后步骤 从核仁到核质，再到其出口到细胞质。 该组装途径需要一个动态的重塑步骤，其中蛋白质蛋白质，RNA-蛋白质和RNA-RNA相互作用得到建立，破坏和重新配置。 人类途径的失调导致许多与细胞生长或增殖（包括癌症）改变有关的疾病。 核糖体生物发生缺陷的酵母突变体的筛选，以及净化核糖体组装中间体和鉴定其成分的方法的开发，导致识别> 170跨性别作用 核糖体组装所需的因素。了解核糖体生物发生机制的核心是找出每个组装因子中每个组装因子所扮演的精确作用。 他们中的哪个接触rnna？ 哪种蛋白质相互作用？一个人可以定义新生RRNP中的组装邻域，如在体外组装的原核生物核糖体所观察到的吗？ 这些因素以什么顺序与前体相关，执行其功能，然后与颗粒分离？通过什么方式，组装因子和核糖体蛋白是募集到前丝体的，然后被激活的，然后从前RRNP中释放的？ 提出了实验来解决这些问题。 我们关注的是成熟60s核糖体亚基的前体成熟的两个特定连续步骤 - 66SA3组装中间体的成熟，然后是66SB颗粒。为了开始建立因素功能机制的范例，我们将解决上述问题 这两个步骤组装所需的几个因素。 此外，我们将更详细地研究死盒蛋白DRS1的作用。该推定的ATPase是否可以通过触发与前RRNA结合并防止其过早裂解的负调节器的释放来使66SB颗粒的成熟？ 高度的真核核糖体生物发生涉及的分子的保守性有望说，在酵母中发现的核糖体生物发生的原理将提供蓝图以研究更多的多样性 后生动物中核糖体组装的调节模式，包括在许多疾病中破坏的核糖体组装方式。