CAREER: GTPase Control of Ribosome Biogenesis

核糖体是一种复杂的大分子机器，由细菌中的至少54种蛋白质和3种RNA分子组成，其组装在所有细胞中都是一个知之甚少的过程。GTP酶在从细菌到人类的生物体中的核糖体生物合成中具有关键作用，但其分子功能尚未得到很好的表征。职业发展计划中概述的研究项目的总体目标是阐明一个重要的GTdR，RbgA，如何参与枯草芽孢杆菌核糖体大亚基的生物合成。RbgA在大多数细菌和真核生物中广泛保守，因此关于RbgA功能所获得的结果将适用于许多系统中的核糖体生物合成。大量的遗传工具、在体外重建细菌核糖体生物发生的能力以及革兰氏阳性细菌在核糖体的合成和生物发生中利用替代途径的事实制造了B。subtilis是研究RbgA功能的理想体系。已经开发了一种模型，其中RbgA通过调节核糖体蛋白L16的插入促进45 S中间体最终成熟为成熟的50 S亚基。为了验证这一工作模型以及RbgA在核糖体组装和亚基偶联中作为具有双重功能的因子的假设，将追求以下具体目标：1.一种新型核糖体大亚基中间体的结构和功能表征。2. RbgA在核糖体生物合成中功能的体内外分析。这项研究的智力价值是更好地了解非核糖体因素如何影响体内核糖体生物合成。作为职业发展计划教育部分的一部分，正在开发一个强调使用各种基因组技术的动手本科实验室课程。本课程将补充现有的本科生讲座和生物信息学课程已经被教授的布里顿博士。本研究项目的主要影响有：1。为本科生的研究生院或生物技术工作的准备，让他们在基因组学，2。目前的研究项目纳入本科实验室课程的整合，3。本科生和研究生的实验室培训; 4.阐明核糖体如何在体内形成，这是细菌、古细菌和真核细胞的重要过程。

The assembly of the ribosome, a complex macromolecular machine consisting of at least 54 proteins and 3 RNA molecules in bacteria, is a poorly understood process in all cells. GTPases have critical roles in ribosome biogenesis in organisms ranging from bacteria to humans, yet their molecular functions are not well characterized. The overall objective of the research project outlined in the career development plan is to elucidate how an essential GTPase, RbgA, participates in the biogenesis of the large ribosomal subunit in Bacillus subtilis. RbgA is widely conserved in most bacteria and eukaryotes and therefore the results obtained regarding RbgA function will be applicable to ribosome biogenesis in many systems. The plethora of genetic tools, the ability to reconstitute bacterial ribosome biogenesis in vitro, and the fact that Gram positive bacteria utilize alternative pathways in the synthesis and biogenesis of the ribosome make B. subtilis an ideal system to study RbgA function. A model in which RbgA facilitates the final maturation of a 45S intermediate into a mature 50S subunit by regulating the insertion of ribosomal protein L16 has been developed. To test this working model and the hypothesis that RbgA serves as a factor with dual functions in ribosome assembly and subunit coupling the following specific aims will be pursued: 1. Structural and functional characterization of a novel large ribosomal subunit intermediate. 2. In vitro and in vivo analysis of the function of RbgA in ribosome biogenesis. The intellectual merit of this research is a better understanding of how non-ribosomal factors influence ribosome biogenesis in vivo. As part of the educational component of the career development plan a hands-on undergraduate laboratory course highlighting the use of various genomic technologies is being developed. This course will supplement an existing undergraduate lecture and bioinformatics course already being taught by Dr. Britton. The broader impacts of this research project are: 1. The preparation of undergraduates for graduate school or jobs in biotechnology by giving them a solid foundation in genomics, 2. The integration of current research projects into the undergraduate laboratory course, 3. The training of undergraduates and graduate students in the laboratory, and 4. Elucidating how ribosomes are formed in vivo, a process important for bacteria, archaea, and eukaryotic cells.