Novel targets within the Hepatitis C Virus nonstructural protein NS4B and their inhibition using distinct classes of small molecules
Abstract/Contents
- Abstract
- Hepatitis C Virus (HCV) is the causative agent of significant liver disease, including cirrhosis and hepatocellular carcinoma. This virus infects greater than 2% of the world's population, and treatment options for these patients are limited; thus, this virus represents a major public health problem. In this thesis, we aim to enhance our ability to solve this problem by unraveling the role that NS4B, one of the HCV nonstructural proteins, plays in the viral life cycle. Drawing on similarities to other plus-sense RNA viruses, we identify an RNA binding activity in NS4B, demonstrate that this activity is specific for the 3' terminus of the HCV genome template, and characterize the domains of NS4B that are responsible for this activity. We utilize microfluidic technology to perform a high-throughput screen for small molecule inhibitors of this RNA binding activity and identify clemizole hydrochloride, among others, as an effective inhibitor, both in vitro and in cell culture. Genetic analysis reveals mutations in NS4B than confer resistance to clemizole, as well as mutations in NS3 that suppress the genetic disruption of NS4B's RNA-binding domain. In addition to clemizole, our work identifies a small molecule inhibitor of HCV that affects a different activity of NS4B, its ability to form membrane-associated foci. We provide genetic, biochemical, and cell biological evidence that NS4B is the target of this drug, and an in vitro light scattering assay further suggests that it is the second amphipathic helix of NS4B that is the target. In total, our results demonstrate that two different activities of NS4B are each a valid pharmacological target for HCV antivirals and uncover two candidate compounds that have potential for further pharmaceutical development.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Copyright date | 2010 |
| Publication date | 2009, c2010; 2009 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Bryson, Paul David |
|---|---|
| Associated with | Stanford University, Department of Microbiology and Immunology. |
| Primary advisor | Glenn, Jeffrey S, 1962- |
| Thesis advisor | Glenn, Jeffrey S, 1962- |
| Thesis advisor | Greenberg, Harry B |
| Thesis advisor | Levy, Shoshana |
| Thesis advisor | Sarnow, P. (Peter) |
| Advisor | Greenberg, Harry B |
| Advisor | Levy, Shoshana |
| Advisor | Sarnow, P. (Peter) |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Paul David Bryson. |
|---|---|
| Note | Submitted to the Department of Microbiology & Immunology. |
| Thesis | Ph.D. Stanford University 2010 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2010 by Paul David Bryson
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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