Narute, Purushottam
(2012)
HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
This is the latest version of this item.
Abstract
Human immunodeficiency virus-1 (HIV-1) is a lentivirus responsible for development of AIDS. In addition to typical retroviral proteins (Gag, Pol and Env), primate lentiviruses like HIV-1 encode two regulatory (Tat and Rev) and four accessory proteins (Vif, Vpu, Vpr and Nef). The HIV-1 accessory factor Nef is essential for high-titer viral replication and AIDS progression. Nef function requires interaction with many host cell proteins, including specific members of the Src kinase family. In this dissertation project, I explored whether Src-family kinase (SFK) activation is a conserved property of nef alleles from a wide range of primary HIV-1 isolates and its sensitivity to selective pharmacological inhibitors. Representative Nef proteins from the major HIV-1 subtypes A1, A2, B, C, F1, F2, G, H, J and K strongly activated Hck and Lyn as well as c-Src to a lesser extent, demonstrating for the first time that SFK activation is a highly conserved property of primary M-group HIV-1 Nef isolates. Moreover, patient-derived Nef proteins also strongly activated Hck. Recently, our group identified 4-amino diphenylfuranopyrimidines (DFPs) and diphenylpyrazolyldiazene (PPD-B9) compounds that selectively inhibit Nef-dependent SFK activation as well as HIV replication. To determine whether these novel compounds exhibit broad-spectrum Nef-dependent antiretroviral activity against HIV-1, I first constructed chimeric forms of the viral strain NL4-3 expressing the same 10 primary nef alleles. The infectivity and replication of these Nef chimeras was indistinguishable from that of wild-type in three distinct cell lines (MT2, U87MG and CEM-T4). Importantly, the 4-aminopropanol and 4-aminobutanol derivatives of DFP as well as PPD-B9 potently inhibited the replication of all chimeric forms of HIV-1 in both U87MG and CEM-T4 cells in a Nef-dependent manner. The effects of these compounds against HIV replication correlated with inhibition of Nef-dependent activation of endogenous SFKs. My results demonstrate that the activation of Hck, Lyn and c-Src by Nef is highly conserved among all major clades of HIV-1 and that selective targeting of this pathway uniformly inhibits HIV-1 replication. My results have strong public health significance for developing therapeutics against current drug resistant variants of HIV-1.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
| Creators | Email | Pitt Username | ORCID  |
|---|
| Narute, Purushottam | pun1@pitt.edu | PUN1 | |
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| ETD Committee: |
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| Date: |
29 June 2012 |
| Date Type: |
Publication |
| Defense Date: |
27 February 2012 |
| Approval Date: |
29 June 2012 |
| Submission Date: |
14 March 2012 |
| Access Restriction: |
5 year -- Restrict access to University of Pittsburgh for a period of 5 years. |
| Number of Pages: |
174 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Public Health > Infectious Diseases and Microbiology |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
HIV-1, Nef, Src family kinase, Hck, Lyn, Src, AIDS, Drug Discovery |
| Date Deposited: |
29 Jun 2012 21:11 |
| Last Modified: |
29 Jun 2017 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/12494 |
Available Versions of this Item
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HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis. (deposited 29 Jun 2012 21:11)
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