eprintid: 12494 rev_number: 28 userid: 651 dir: disk0/00/01/24/94 datestamp: 2012-06-29 21:11:27 lastmod: 2017-06-29 05:15:07 status_changed: 2012-06-29 21:11:27 type: thesis_degree succeeds: 11444 metadata_visibility: show contact_email: purushottam.narute@gmail.com item_issues_id: duplicate_title_11444 item_issues_id: thesis_degree_versioning item_issues_type: duplicate_title item_issues_type: thesis_degree_versioning item_issues_description: Duplicate title to Narute, Purushottam HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis. Doctoral Dissertation, University of Pittsburgh. item_issues_description: ETD 12494 is using versioning. item_issues_timestamp: 2012-06-22 06:10:59 item_issues_timestamp: 2013-09-11 17:48:34 item_issues_status: autoresolved item_issues_status: discovered item_issues_count: 1 eprint_status: archive creators_name: Narute, Purushottam creators_email: pun1@pitt.edu creators_id: PUN1 title: HIV-1 Nef-Src Family Kinase Interaction: A Novel Target for the Inhibition of HIV-1 Pathogenesis ispublished: unpub divisions: sch_gsph_infectiousdiseasesmicrobiology full_text_status: public keywords: HIV-1, Nef, Src family kinase, Hck, Lyn, Src, AIDS, Drug Discovery 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. date: 2012-06-29 date_type: published pages: 174 institution: University of Pittsburgh refereed: TRUE etdcommittee_type: thesis_advisor etdcommittee_type: committee_member etdcommittee_type: committee_member etdcommittee_type: committee_member etdcommittee_name: Smithgall, Thomas etdcommittee_name: Reinhart, Todd etdcommittee_name: Wang, Tianyi etdcommittee_name: Homa, Fred etdcommittee_email: tsmithga@pitt.edu etdcommittee_email: reinhar@pitt.edu etdcommittee_email: tywang@pitt.edu etdcommittee_email: flhoma@pitt.edu etdcommittee_id: TSMITHGA etdcommittee_id: REINHAR etdcommittee_id: TYWANG etdcommittee_id: FLHOMA etd_defense_date: 2012-02-27 etd_approval_date: 2012-06-29 etd_submission_date: 2012-03-14 etd_release_date: 2012-06-29 etd_access_restriction: 5_year etd_patent_pending: FALSE assigned_doi: doi:10.5195/pitt.etd.2012.11444 thesis_type: dissertation degree: PhD citation: 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) document_url: http://d-scholarship-dev.library.pitt.edu/12494/2/Narute_PS_ETD.pdf