CAREER: Investigation of RNA Unwinding And Ribosomal Frameshifting

智力优势：核糖体是一种存在于所有生物体中的大分子机器，主要功能是产生蛋白质。 这一过程被称为翻译，非常复杂，并受到许多辅助因子的高度调节，这些辅助因子有助于有效的蛋白质生产。 在正常的翻译中，核糖体通过将信息（RNA）上的三个位置翻译成一个氨基酸来翻译信息。 通过一次推进三个位置，核糖体继续构建蛋白质序列，直到遇到称为终止密码子的信息上的特定序列，此时全长蛋白质从核糖体释放。 通常，复杂的三维结构存在于信息中，并且必须展开以继续翻译。 在某些情况下，这些结构可以诱导核糖体滑回一个位置。 这导致核糖体处于不同的框或移码中，并将导致核糖体跳过在原始框中遇到的终止密码子，通常产生更长的蛋白质序列。 虽然我们对移码有很多了解，但仍然有一些重要的问题需要详细解决。 在本项目中，将采用先进的生物物理技术解决与移码有关的下列问题。 首先，将阐明核糖体解开信息中复杂的三维结构以继续翻译的机制。 其次，这些结构如何影响核糖体内的整体和局部运动将被研究。 最后，为什么有些结构有更大的倾向，诱导移框比别人通过探测固有的动态运动的一个子集的结构，不同的移框效率将被调查。 这些实验将提供一个全面的描述展开，并导致一个更大的描述框架转移的过程。 此外，这些实验将对核糖体和生物物理学领域产生深远的影响，并通过推动生物物理技术的极限和能力对科学界产生普遍影响，特别是在密苏里州大学。美国只有5.5%的人口从事科学、技术、工程和数学（STEM）工作，重要的是，必须强调对公众以及公立学校和高等教育机构的教育和认识。 已承诺制定和执行若干具体活动，这些活动将对若干领域产生直接和立即的影响，特别强调代表性不足和收入较低的人口。 教育和推广活动包括：协助一个MU教师培训计划，旨在为K-8年级;制定新的课程，以扩大教师培训计划，包括高中教师在化学和生物化学，目的是将尖端科学融入课堂;提供本科研究机会，以高成绩的学生和少数民族学生准备他们的职业生涯在干;继续指导研究生，并鼓励推广活动;从事公开演讲的机会，在附近地区，以及在大学与最低限度的研究机会;并制定本科和研究生水平的课程材料，将挑战，并提供在科学研究的实践经验。

Intellectual Merit: The ribosome is a large, macromolecular machine found in all organisms that primarily functions to produce protein. This process, referred to as translation, is very complex and highly regulated with many accessory factors that facilitate efficient protein production. In normal translation, the ribosome translates a message by interpreting three positions on the message (RNA) into one amino acid. By advancing three positions at a time, the ribosome continues to build the protein sequence until a specific sequence on the message known as a stop codon is encountered at which point the full length protein is released from the ribosome. Often, complex three dimensional structures are present within the message and must be unwound for translation to continue. In some cases, these structures can induce the ribosome to slip back one position. This causes the ribosome to be in a different frame or frameshift and will cause the ribosome to skip over the stop codon that would have been encountered in the original frame generally producing a longer protein sequence. While much is known about frameshifting, there are still important questions that remain to be addressed in detail. In this project, the following questions related to frameshifting will be addressed by employing advanced biophysical techniques. First, the mechanism by which the ribosome unwinds complex three dimensional structures within the message in order to continue translation will be elucidated. Second, how these structures influence the global and local motions within the ribosome will be investigated. Finally, why some structures have a greater propensity to induce frameshifting than others by probing the inherent dynamic motions of a subset of structures that vary in frameshifting efficiency will be investigated. These experiments will provide a comprehensive description of unwinding and lead to a greater description of the process of frameshifting. Further, these experiments will have a profound impact on the ribosome and biophysics fields as well as a general impact on the scientific community particularly at the University of Missouri by pushing the limits and capabilities of biophysical techniques.Broader Impact: With only 5.5% of the population in science, technology, engineering, and mathematics (STEM) in the United States, it is essential that education and awareness is emphasized for both the general public and in our public schools and higher education institutions. A committment to develop and implement several specific activities that will have a direct and immediate impact in several areas with a special emphasis on underrepresented and lower income populations has been made. Education and outreach activities include: to assist with an MU teacher training program intended for grades K-8; to develop new curricula to expand the teacher training program to include high school teachers in chemistry and biochemistry with the intention of integrating cutting edge science into the classroom; to provide undergraduate research opportunities to high achieving students and to minority students preparing them for careers in STEM; to continue mentoring graduate students and encourage outreach activities; to engage in public speaking opportunities in the immediate area as well as in Universities with minimal research opportunities; and to develop undergraduate and graduate level course material that will challenge and provide hands on experience in scientific research.