eprintid: 14146 rev_number: 17 userid: 1346 importid: 661 dir: disk0/00/01/41/46 datestamp: 2012-09-13 18:06:59 lastmod: 2019-01-26 10:55:07 status_changed: 2012-09-13 18:06:59 type: article metadata_visibility: show contact_email: tsmithga@pitt.edu item_issues_count: 0 eprint_status: archive creators_name: Narute, PS creators_name: Smithgall, TE creators_email: creators_email: tsmithga@pitt.edu creators_id: creators_id: TSMITHGA contributors_type: http://www.loc.gov/loc.terms/relators/EDT contributors_name: Vartanian, Jean-Pierre title: Nef alleles from all major HIV-1 clades activate Src-family kinases and enhance HIV-1 replication in an inhibitor-sensitive manner ispublished: pub divisions: sch_gsph_infectiousdiseasesmicrobiology divisions: sch_med_Microbiology_Molecular_Genetics full_text_status: public abstract: 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. Here we explored whether Src-family kinase activation is a conserved property of Nef alleles from a wide range of primary HIV-1 isolates and their 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 Src-family kinase activation is a highly conserved property of primary M-group HIV-1 Nef isolates. Recently, we identified 4-amino substituted diphenylfuropyrimidines (DFPs) that selectively inhibit Nef-dependent activation of Src-family kinases as well as HIV replication. To determine whether DFP compounds exhibit broad-spectrum Nef-dependent antiretroviral activity against HIV-1, we first constructed chimeric forms of the HIV-1 strain NL4-3 expressing each of the primary Nef alleles. The infectivity and replication of these Nef chimeras was indistinguishable from that of wild-type virus in two distinct cell lines (U87MG astroglial cells and CEM-T4 lymphoblasts). Importantly, the 4-aminopropanol and 4-aminobutanol derivatives of DFP potently inhibited the replication of all chimeric forms of HIV-1 in both U87MG and CEM-T4 cells in a Nef-dependent manner. The antiretroviral effects of these compounds correlated with inhibition of Nef-dependent activation of endogenous Src-family kinases in the HIV-infected cells. Our 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. © 2012 Narute, Smithgall. date: 2012-02-29 date_type: published publication: PLoS ONE volume: 7 number: 2 refereed: TRUE id_number: 10.1371/journal.pone.0032561 other_id: NLM PMC3290594 pmcid: PMC3290594 pmid: 22393415 mesh_headings: Alleles mesh_headings: Amino Acid Sequence mesh_headings: Anti-HIV Agents--pharmacology mesh_headings: Cell Line, Tumor mesh_headings: Dimerization mesh_headings: HIV Infections mesh_headings: HIV-1--genetics mesh_headings: Humans mesh_headings: Molecular Conformation mesh_headings: Molecular Sequence Data mesh_headings: Proto-Oncogene Proteins c-hck--metabolism mesh_headings: Sequence Analysis, DNA mesh_headings: Sequence Homology, Amino Acid mesh_headings: Species Specificity mesh_headings: Virus Replication mesh_headings: nef Gene Products, Human Immunodeficiency Virus--genetics mesh_headings: nef Gene Products, Human Immunodeficiency Virus--metabolism mesh_headings: src-Family Kinases--metabolism chemical_names: Anti-HIV Agents chemical_names: nef Gene Products, Human Immunodeficiency Virus chemical_names: nef protein, Human immunodeficiency virus 1 chemical_names: HCK protein, human chemical_names: Proto-Oncogene Proteins c-hck chemical_names: lyn protein-tyrosine kinase chemical_names: src-Family Kinases citation: Narute, PS and Smithgall, TE (2012) Nef alleles from all major HIV-1 clades activate Src-family kinases and enhance HIV-1 replication in an inhibitor-sensitive manner. PLoS ONE, 7 (2). document_url: http://d-scholarship-dev.library.pitt.edu/14146/1/Nef_Alleles.pdf document_url: http://d-scholarship-dev.library.pitt.edu/14146/8/licence.txt