Advisor(s)
Chad Petit
Committee Member(s)
David Schneider
Todd Green
William Britt
William Placzek
School
Joint Health Sciences (Interdisciplinary)
Document Type
Dissertation
Department (new version)
Biochemistry and Molecular Genetics
Date of Award
9-11-2025
Abstract
Virulence factors are an important mechanism by which viruses combat the host immune system, therefore a thorough understanding of the workings of these proteins will be of significant value in prevention of disease associated with viral infections. In this dissertation we investigate the main virulence factors of two viruses, the Influenza A Virus (IAV) and SARS-CoV-2, and their interactions with host proteins, providing critical information for structure-based drug design. In chapter one we studied the nonstructural protein 1 (NS1) of IAV and its interaction with the 30-kda subunit of the cleavage and polyadenylation specificity factor (CPSF30) to inhibit host cell translation. We find that a naturally occurring mutation in NS1, D171A, has several effects on both NS1 as a whole and its interaction with CPSF30. We observe decreased binding affinity to CPSF30, a difference in the n-value of this interaction indicating changes in either stoichiometry or percent active population, and an increase in NS1 stability during denaturation studies. Through these studies we also uncover novel findings about NS1 itself including the presence of an intermediate state during denaturation and a trimeric complex with CPSF30 which differs from the literature. In chapter two we studied the nonstructural protein 1 of SARS-CoV-2 (S2-nsp1) and its interaction with the host cell immunophilin Cyclophilin A (CyPA), whose inhibitor has been shown to attenuate coronavirus replication. We found that this interaction is conserved from SARS-CoV and mapped the interaction between the two proteins from both sides using nuclear magnetic resonance (NMR) chemical shift perturbation (CSP). We successfully identified key residues involved in the interaction on both S2-nsp1 and CyPA and used our findings to generate a model of the complex. Taken together, our findings shed light onto crucial proteins for both of these viruses and provide structural information that will prove useful in creating novel compounds or modifying existing ones to counteract their effects on the immune system.
ProQuest ID
Recommended Citation
Blake, Morgan E., "Virulence Factors And Their Binding Partners: A Biochemical Study" (2025). All ETDs from UAB. 7396.
https://digitalcommons.library.uab.edu/etd-collection/7396
