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.
