While the proportion of people infected with the Human Immunodeficiency Virus (HIV) dying of Acquired Immunodeficiency Syndrome (AIDS) has decreased significantly over the past four decades as antiretroviral therapy has become more common, the full structure of the HIV-1 genome remains unsolved. The 5′ leader of the RNA sequence is of particular interest to solve, as it is the most highly conserved region of HIV’s genome and plays a key role in the packaging and proliferation of the virus. NMR spectroscopy is our lab’s method of elucidating RNA complexes due to its ability to collect data on dynamic complexes in solution. Our group will be using a U1A reporter protein paramagnetically tagged by M8-DOTA-SPy to induce long-range pseudocontact shifts (PCS) in the NMR spectra of our RNA construct. The goal of this project is to elucidate the tertiary structure of the orientation of the TAR and polyA stems of the 5′ leader in relation to each other to build on the Brown et. al paper published in 2020. We will show binding through electrophoretic mobility shift assays (EMSAs) and show PCS of a TAR polyA construct.
Student support provided by the National Institute of General Medical Sciences, National Institutes of Health (NIGMS/NIH) under National Research Service Award T34 GM 136497, NIGMS Initiative for Maximizing Student Development Grant (2R25-GM55036), NIAID grant ##8 R01 AI150498, and the Howard Hughes Medical Institute.