Assembling an understanding of Ribosome Biogenesis

描述(申请人提供)：摘要/摘要：核糖体是多组分细胞RNPs中最普遍、最保守和最具特征的一种。虽然人们对核糖体的结构和功能了解很多，但仍有许多细节需要阐明，特别是关于核糖体的组装及其对核糖体功能能力的影响。因此，我们研究大肠杆菌小核糖体亚基(SSU)的生物合成不仅将对理解这些颗粒的组装和功能具有重要影响，而且将对其他RNP的动态组装和功能产生重要影响。此外，大约40%的大肠杆菌总能量生产被消耗在合成核糖体成分上。这一观察结果以及大肠杆菌生长速度和核糖体生物发生之间的直接相关性表明，准确组装和功能的核糖体对细胞活力至关重要。我们的长期目标是详细了解核糖体的生物发生，从而揭示这一动态过程对细胞生理的基本方面的影响，如生长调节。本提案详细介绍了大肠杆菌SSU的生物发生研究。这项工作的目的是深入了解在功能SSU组装过程中的构象变化和调节这些变化的因素。已经提出了三个具体目标来解决这些问题。首先，将确定16S rRNA和特定rRNA核苷酸和核糖体蛋白中的构象变化，这些变化有助于在体外和体内组装的不同阶段发生这些变化。其次，利用遗传学和生物化学的结合，新的SSU组装中间体将被识别和表征。第三，分析了促进SSU组装的辅助因素。将详细探讨本供资期间确定的因素，并确定和说明其他因素。我们在当前资金周期中的进展和我们的初步数据表明，所有三个特定目标都将成功，因此将直接影响对SSU生物发生的理解，并极大地扩大对RNA-蛋白质相互作用和RNP组装的总体理解。通过揭示对SSU的组装和功能都至关重要的方面，我们可以确定新的抗菌作用的靶点。鉴于抗药性细菌菌株的日益流行，发现这样的靶点对人类健康至关重要。

DESCRIPTION (provided by applicant): Summary/Abstract: The ribosome is one the most ubiquitously conserved and best characterized of the multi- component cellular RNPs. While much is known about ribosome structure and function, there are still many details to be elucidated especially regarding ribosome assembly and its impact the functional capacity of ribosomes. Thus, our work to address biosynthesis of E. coli small ribosomal subunits (SSUs) will have a significant impact on understanding not only assembly and function of these particles but also on the dynamic assembly and function of other RNPs. Moreover approximately 40% of the total energy production of E. coli is consumed to synthesize ribosomal components. This observation and the direct correlation between the rates of E. coli growth and ribosome biogenesis indicate that an accurately assembled and functional ribosome is of utmost importance for cell viability. Our long term goal is to gain a detailed understanding of ribosome biogenesis and thus reveal the impact of this dynamic process on fundamental aspects of cell physiology, such as growth regulation. This proposal details studies of E. coli SSU biogenesis. The purpose of this work is to gain an in-depth understanding of conformational changes and the factors that modulate these changes during the course of functional SSU assembly. Three specific aims have been proposed to address these questions. First, conformational changes within 16S rRNA and specific rRNA nucleotides and ribosomal proteins that facilitate these changes during different stages of assembly in vitro and in vivo will be identified. Second, using a combination of genetics and biochemistry, novel SSU assembly intermediates will be identified and characterized. Third, auxiliary factors that facilitate SSU assembly will be analyzed. Factors identified in the current funding period will be explored in detail and additional factors will be identified and characterized. Our progress during the current funding cycle and our preliminary data indicate that all three specific aims will be successful and thus will directly impact understanding of SSU biogenesis and greatly expand understanding of RNA-protein interactions and RNP assembly in general. By revealing aspects that are critical for both assembly and function of SSUs, we may identify novel target sites for antimicrobial action. Given the growing prevalence of antibiotic resistant bacterial strains, the discovery of such targets is critical for human health.