Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form

HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis

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 (Unpublished)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.

[img]
Preview
PDF
Primary Text

Download (3MB) | Preview

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.


Share

Citation/Export:
Social Networking:
Share |

Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Narute, Purushottampun1@pitt.eduPUN1
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorSmithgall, Thomastsmithga@pitt.eduTSMITHGA
Committee MemberReinhart, Toddreinhar@pitt.eduREINHAR
Committee MemberWang, Tianyitywang@pitt.eduTYWANG
Committee MemberHoma, Fredflhoma@pitt.eduFLHOMA
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: Graduate 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

  • HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis. (deposited 29 Jun 2012 21:11) [Currently Displayed]

Metrics

Monthly Views for the past 3 years

Plum Analytics


Actions (login required)

View Item View Item