Title: Understanding HIV reverse transcription: the structure and function of the N-terminal domain in human Lysyl-tRNA Synthetase Author: Tran, U., Nguyen, H., Refaei, M., Calhoun, J., and Tsang, P. College of Arts & Sciences, University of Cincinnati

Abstract

By Uyen Tran, Biochemistry

Advisor: Pearl Tsang

Presentation ID: PM_D19

Abstract: The enzyme Lysyl-tRNA Synthetase in human, abbreviated hKRS, is packaged into new HIV-1 virus particles. The eukaryotic enzyme consists of three domains: an anticodon binding domain, a catalytic domain, and an N-terminal domain that is unique to higher eukaryotic organisms. Previous studies have shown that the presence of the N-terminal domain (NTD) greatly increases the selective incorporation of tRNA-Lys3 of HIV-1; however, the NTD structure and the means by which it interacts with the tRNA are still not well characterized or understood. The research conducted and described here involves purification of hKRS NTD and our studies of its structural properties. This specifically involved the growth and purification of the recombinant form of the NTD in Escherichia coli BL21 DE3 cells. Purified NTD protein that was obtained was then concentrated and the protein concentration was determined using UV spectrophotometry and other techniques. The purified, concentrated NTD was then studied using Circular Dichroism to examine the secondary structure of this protein as a function of concentration and temperature.