4.7 Article

High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction

Journal

Publisher

MDPI
DOI: 10.3390/ijms22179308

Keywords

HIV-1; Vpu; BST-2; NanoLuc Binary Technology; high-throughput screening assay

Funding

  1. National Natural Science Foundation of China [81760783, 21625501, 21936001]
  2. Beijing Outstanding Young Scientist Program [BJJWZYJH01201910005017]

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BST-2 is a host restriction factor that suppresses the release of enveloped viruses by tethering viral particles to the cell surface. Inhibiting the interaction between Vpu and BST-2 could provide a promising strategy for anti-HIV therapy.
Bone marrow stromal cell antigen 2 (BST-2), also known as CD317 or tetherin, has been identified as a host restriction factor that suppresses the release of enveloped viruses from host cells by physically tethering viral particles to the cell surface; however, this host defense can be subverted by multiple viruses. For example, human immunodeficiency virus (HIV)-1 encodes a specific accessory protein, viral protein U (Vpu), to counteract BST-2 by binding to it and directing its lysosomal degradation. Thus, blocking the interaction between Vpu and BST-2 will provide a promising strategy for anti-HIV therapy. Here, we report a NanoLuc Binary Technology (NanoBiT)-based high-throughput screening assay to detect inhibitors that disrupt the Vpu-BST-2 interaction. Out of more than 1000 compounds screened, four inhibitors were identified with strong activity at nontoxic concentrations. In subsequent cell-based BST-2 degradation assays, inhibitor Y-39983 HCl restored the cell-surface and total cellular level of BST-2 in the presence of Vpu. Furthermore, the Vpu-mediated enhancement of pesudotyped viral particle production was inhibited by Y-39983 HCl. Our findings indicate that our newly developed assay can be used for the discovery of potential antiviral molecules with novel mechanisms of action.

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