Transcript – Genetic Studies of an Essential NTPase Motor System in Pathogenic Parasites & Computational Approaches in Drug Discovery

Hi, my name is Joshua Meehan, I’m a fourth year Ph.D. student in the genetics program, I’m here to talk about our work funded by the T3 award given to Dr. Cruz-Reyes who is acting as leader of this project, as well as Dr. Meek, both of which come from the Department of Biochemistry and Biophysics, who are also working with Dr. Thomas in the Department of Chemical Engineering. Together, we are combining our knowledge of molecular parasitology, enzyme kinetics and computational drug development with the overall goal of tackling pathogenic parasites known as Trypanosoma. These parasites cause Chagas disease in humans, in agricultural animals, in Latin America and the southern US, as well as sleeping sickness or African trypanosomiasis in sub-Saharan Africa. The World Health Organization considers these and other related parasites as neglected diseases, which require further investigation. We are focusing on a unique biological process known as RNA editing. Maturation of the RNA and parasites is needed to create functional sequences that can be used in translation of proteins. This essential RNA editing pathway is very error prone and blocking the process using either genetic manipulation or candidate inhibitor compounds could be used to inhibit the growth of the parasite and thereby the disease is caused by them. However, there is a need to increase our understanding of the editing mechanisms as not a whole lot is known about the source of editing fidelity and error. And we will then use that data to develop specific compounds against the editing machinery. The editing machinery is formed by subsets of proteins of editing complexes. One of these editing complexes discovered in the cruise raised lab is a molecular motor that is fueled by hydrolysis of ATP, an energy rich molecule found inside the parasite cells. This editing complex contains an enzyme which remodels RNA structures as well as a zinc finger containing cofactors protein known as H2 EF1.

We utilize three high through methods to uncover more about this editing complex cross-linking. Mass spectrometry of the protein complex revealed that the cofactors protein causes confirmation of changes to our age to which provide greater stability to the enzyme during editing genetic depletion of these proteins, followed by RNA sequences they target so that this complex reduces error in the editing process and is essential for promoting proper RNA maturation. Finally, a novel DM’s map approach was developed to investigate the change in RNA structure throughout the editing process so that we may see specific changes that the remodeling enzyme promotes or inhibits. Using this information, we were able to model the ATP binding side of this enzyme and through this modeling and simulations, came up with several candidate compounds which are expected to inhibit the editing machinery and increase the error rate of editing, killing the parasites. The funding provided for this work has resulted in four peer-reviewed publications from the groups involved, as well as providing essential data for writing grant proposals seeking extramural funding. Thank you for visiting my poster.