CAREER: Dissecting Inter-domain Communication in Methionyl-tRNA Synthetase

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This project's ultimate goal is to explain protein-RNA conformational dynamics.For efficient catalysis, information from the Escherichia coli methionyl-tRNA synthetase (MetRS) anticodon binding domain must be transmitted to a remote active site upon interaction with the enzyme's cognate tRNA. Biochemical evidence suggests this signaling event requires changes in both RNA and protein structure. Such long-range conformational changes are probably very common in macromolecular machines, such as ribosomes and spliceosomes, and, therefore, of fundamental biological importance, yet they remain virtually unexplored. The MetRS model system, amenable to detailed, quantitative analysis, will help to remedy this gap. Mutagenetic, kinetic, and spectroscopic approaches will be used to investigate how a specific peptide component at the domain interface of MetRS contributes to inter-domain communication. In addition, crystallography will be used to observe structural differences between MetRS in complex with its cognate tRNA and substrate analogs and with unliganded enyzme. Results obtained in this interdisciplinary study will contribute to a new understanding of inter-domain communication in aminoacyl-tRNA synthetases and to general knowledge about signaling mechanisms in enzymes with distant ligand-binding and catalytic active sites.Broader Impacts: The project will engage undergraduate and graduate education at the interface of biology, chemistry, and physics at Wake Forest University. The PI carries her knowledge of, and enthusiasm for, science research to ongoing outreach programs in local public schools. The biotechnology focus of these teacher workshops and student enrichment activities strengthens local efforts to motivate and to train rural and urban high school graduates to pursue postsecondary science careers. These efforts enhance both the PI's career development in research and education and the community's scientific awareness and training with an eye toward economic growth.