Assembling an understanding of Ribosome Biogenesis

描述（由申请人提供）：概述/摘要：核糖体是多组分细胞RNP中最普遍保守和最好表征的一种.虽然对核糖体的结构和功能已经有了很多了解，但仍然有许多细节需要阐明，特别是关于核糖体组装及其对核糖体功能能力的影响。因此，我们的工作，以解决生物合成的E。大肠杆菌小核糖体亚基（SSU）的研究不仅对理解这些颗粒的组装和功能，而且对理解其他RNP的动态组装和功能具有重要影响。此外，约40%的E.大肠杆菌被消耗以合成核糖体组分。这一观察结果与E.大肠杆菌生长和核糖体生物发生表明，一个准确组装和功能性的核糖体是细胞活力的最重要的。我们的长期目标是详细了解核糖体生物发生，从而揭示这种动态过程对细胞生理学基本方面的影响，如生长调节。 本文详细介绍了E. coli SSU生物合成。这项工作的目的是深入了解功能性SSU组装过程中的构象变化和调节这些变化的因素。为解决这些问题提出了三个具体目标。首先，16 S rRNA和特定rRNA核苷酸和核糖体蛋白内的构象变化，促进这些变化在体外和体内组装的不同阶段将被确定。其次，使用遗传学和生物化学的组合，新的SSU组装中间体将被识别和表征。第三，辅助因素，促进SSU组装将进行分析。将详细探讨在本供资期间查明的因素，并查明和描述其他因素。我们在当前资金周期中的进展和我们的初步数据表明，所有三个具体目标都将成功，从而将直接影响对SSU生物起源的理解，并大大扩展对RNA-蛋白质相互作用和RNP组装的理解。通过揭示对SSU的组装和功能都至关重要的方面，我们可以确定抗菌作用的新靶点。鉴于抗生素耐药菌株的日益流行，发现此类靶标对人类健康至关重要。 公共卫生相关性：项目叙述：已知的抗生素具有共同的靶标和化学部分，并且鉴于多重耐药细菌的流行，鉴定其他抗生素和靶标对人类健康至关重要。细菌核糖体的生物合成为抗菌药物的开发提供了新的前景，因此我们对大肠杆菌核糖体的生物合成进行了研究。大肠杆菌的研究可能导致抗生素作用的新靶点的鉴定，并可能有助于缓解作为健康问题的细菌耐药性问题。

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. PUBLIC HEALTH RELEVANCE: Project Narrative: Known antibiotics share common targets and chemical moieties and given the prevalence of multi-drug resistant bacteria, the identification of additional antibiotics and targets is critical for human health. It has been proposed that bacterial ribosome biogenesis represents promising new ground for development of antimicrobials and thus our studies of ribosome biogenesis in E. coli may result in the identification of novel targets for antibiotic action and may aid in alleviating the problem of bacterial drug resistance as a health concern.