The influence of mRNA secondary structures on ribosome conformational dynamics
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[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Synthesizing proteins in the cell is a critical aspect of life. Protein synthesis is a complicated process and involve highly functional machines at a molecular level. The ribosome is the molecular machine that translate coded sequences of nucleic acids into functional proteins. Understanding how ribosomes function is key to understanding protein synthesis / translation. We focus our work on ribosomes from bacterial cells. This allows us to study much simpler systems and extrapolate our knowledge to higher levels. One key challenge in the field is to be able to isolate a high quantity of good and active ribosomes out of the cell to study it in a controlled environment. Classically known methods involve extensive resources, high technical expertise, and a week of preparation. We developed a one-step protocol to purify ribosomes that are more active than the ones purified from classical methods. This developed technique saves time and money and results in much higher amounts of product. This approach also makes the technique approachable to a wider community of scientists and researchers. The same methodology could be applied towards purifying other molecular machines in the cells. Using these ribosomes, we wanted to investigate how the ribosomes function in cells when faced with specific signals. These signals are utilized by the cells to control protein synthesis. However, in dome diseased cells and for some viruses, normal protein synthesis is overridden by the invaders to produce faulty proteins that could result in a wide range of diseases such as Alzheimer and others. In this study, we investigated how the ribosome functions in the presence of such signals and how close do they need to be to the ribosome to affect protein synthesis. This allows us to design drugs to mimic or inhibit such changes thus fixing faulty protein production or sometimes induce it to inhibit protein synthesis in bacterial cells and as such designing and producing novel drugs.