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Discovery of a diaminoquinoxaline benzenesulfonamide antagonist of HIV-1 Nef function using a yeast-based phenotypic screen

Trible, RP and Narute, P and Emert-Sedlak, LA and Alvarado, JJ and Atkins, K and Thomas, L and Kodama, T and Yanamala, N and Korotchenko, V and Day, BW and Thomas, G and Smithgall, TE (2013) Discovery of a diaminoquinoxaline benzenesulfonamide antagonist of HIV-1 Nef function using a yeast-based phenotypic screen. Retrovirology, 10 (1).

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Abstract

Background: HIV-1 Nef is a viral accessory protein critical for AIDS progression. Nef lacks intrinsic catalytic activity and binds multiple host cell signaling proteins, including Hck and other Src-family tyrosine kinases. Nef binding induces constitutive Hck activation that may contribute to HIV pathogenesis by promoting viral infectivity, replication and downregulation of cell-surface MHC-I molecules. In this study, we developed a yeast-based phenotypic screen to identify small molecules that inhibit the Nef-Hck complex. Results: Nef-Hck interaction was faithfully reconstituted in yeast cells, resulting in kinase activation and growth arrest. Yeast cells expressing the Nef-Hck complex were used to screen a library of small heterocyclic compounds for their ability to rescue growth inhibition. The screen identified a dihydrobenzo-1,4-dioxin-substituted analog of 2-quinoxalinyl-3-aminobenzene-sulfonamide (DQBS) as a potent inhibitor of Nef-dependent HIV-1 replication and MHC-I downregulation in T-cells. Docking studies predicted direct binding of DQBS to Nef which was confirmed in differential scanning fluorimetry assays with recombinant purified Nef protein. DQBS also potently inhibited the replication of HIV-1 NL4-3 chimeras expressing Nef alleles representative of all M-group HIV-1 clades.Conclusions: Our findings demonstrate the utility of a yeast-based growth reversion assay for the identification of small molecule Nef antagonists. Inhibitors of Nef function discovered with this assay, such as DQBS, may complement the activity of current antiretroviral therapies by enabling immune recognition of HIV-infected cells through the rescue of cell surface MHC-I. © 2013 Trible et al.; licensee BioMed Central Ltd.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Trible, RP
Narute, P
Emert-Sedlak, LAlae9@pitt.eduLAE9
Alvarado, JJjja24@pitt.eduJJA24
Atkins, K
Thomas, LTHOMASLA@pitt.eduTHOMASLA
Kodama, T
Yanamala, N
Korotchenko, V
Day, BW
Thomas, Gthomasg@pitt.eduTHOMASG
Smithgall, TEtsmithga@pitt.eduTSMITHGA
Date: 14 November 2013
Date Type: Publication
Journal or Publication Title: Retrovirology
Volume: 10
Number: 1
DOI or Unique Handle: 10.1186/1742-4690-10-135
Schools and Programs: School of Medicine > Microbiology and Molecular Genetics
School of Medicine > Structural Biology
School of Pharmacy > Pharmaceutical Sciences
Refereed: Yes
Date Deposited: 02 Dec 2016 18:24
Last Modified: 30 Oct 2018 12:57
URI: http://d-scholarship.pitt.edu/id/eprint/29644

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