Retroviral-RNA Structure and Function: Investigating the role of aminoacyl-tRNA synthetases and retroviral-RNA structural elements in the initiation of reverse transcription
Year: 2016
Type: dissertation
Abstract: Most human retroviruses are not known to be pathogenic. In fact, the only two retroviruses that have been shown to cause human disease are human immunodeficiency virus (HIV), and human T-cell lymphotropic virus type 1 (HTLV-1)—the causes of acquired immunodeficiency syndrome (AIDS), and adult T-cell leukemia/lymphoma (ATL) among other diseases, respectively. The hallmark of a retrovirus is reverse transcription (RT), the process through the which RNA genome is copied from ssRNA to dsDNA. However, for RT to initiate, a primer must specifically anneal to the primer binding site (PBS) in the 5′ untranslated region (5′UTR) of the retroviral genome. HIV-1 and HTLV-1 use tRNALys3 and tRNAPro to primer RT respectively. Our lab has demonstrated that human lysyl-tRNA synthetase (LysRS) facilitates priming in HIV-1 by binding a tRNA-like-element (TLE) near the PBS, localizing the tRNALys3 primer. As the primer for HTLV-1 RT is tRNAPro, we hypothesized that this virus may employ a similar mechanism involving human glutamyl-prolyl-tRNA synthetase (EPRS) binding to a TLE. Chapter 1 of this thesis will discuss the optimization of RNA probing experiments to study these retroviral RNAs, improvements made to the processing of RNA probing data analyzed by capillary electrophoresis, and a user friendly tool under development to facilitate this processing. Chapter 2 will discuss the probing of the LysRS:HIV-1 interaction, and small angle X-ray scattering studies aimed to understand more about the structure and function of the HIV-1 5’UTR. Finally, Chapter 3 will discuss investigations of the putative interaction between elements within the HTLV-1 5′UTR and EPRS. Using a wide variety of biochemical techniques, these studies aspire to gain information about these viral processes that could eventually aid in the production of therapeutic agents.
Author: Joshua Hatterschide
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2415797660