High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction
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 m...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-08-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/22/17/9308 |
| _version_ | 1797521394252120064 |
|---|---|
| author | Boye Li Xiaoxiao Dong Wenmei Zhang Tian Chen Boyang Yu Wenyue Zhao Yishu Yang Xiaoli Wang Qin Hu Xiayan Wang |
| author_facet | Boye Li Xiaoxiao Dong Wenmei Zhang Tian Chen Boyang Yu Wenyue Zhao Yishu Yang Xiaoli Wang Qin Hu Xiayan Wang |
| author_sort | Boye Li |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T08:11:59Z |
| format | Article |
| id | doaj.art-b8ae1a6b23184553be84ef178032050a |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T08:11:59Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b8ae1a6b23184553be84ef178032050a2023-11-22T10:41:19ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-012217930810.3390/ijms22179308High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein InteractionBoye Li0Xiaoxiao Dong1Wenmei Zhang2Tian Chen3Boyang Yu4Wenyue Zhao5Yishu Yang6Xiaoli Wang7Qin Hu8Xiayan Wang9The Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaThe Faculty of Environment and Life, Beijing University of Technology, Beijing 100124, ChinaBone 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.https://www.mdpi.com/1422-0067/22/17/9308HIV-1VpuBST-2NanoLuc Binary Technologyhigh-throughput screening assay |
| spellingShingle | Boye Li Xiaoxiao Dong Wenmei Zhang Tian Chen Boyang Yu Wenyue Zhao Yishu Yang Xiaoli Wang Qin Hu Xiayan Wang High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction International Journal of Molecular Sciences HIV-1 Vpu BST-2 NanoLuc Binary Technology high-throughput screening assay |
| title | High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction |
| title_full | High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction |
| title_fullStr | High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction |
| title_full_unstemmed | High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction |
| title_short | High-Throughput NanoBiT-Based Screening for Inhibitors of HIV-1 Vpu and Host BST-2 Protein Interaction |
| title_sort | high throughput nanobit based screening for inhibitors of hiv 1 vpu and host bst 2 protein interaction |
| topic | HIV-1 Vpu BST-2 NanoLuc Binary Technology high-throughput screening assay |
| url | https://www.mdpi.com/1422-0067/22/17/9308 |
| work_keys_str_mv | AT boyeli highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT xiaoxiaodong highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT wenmeizhang highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT tianchen highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT boyangyu highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT wenyuezhao highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT yishuyang highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT xiaoliwang highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT qinhu highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction AT xiayanwang highthroughputnanobitbasedscreeningforinhibitorsofhiv1vpuandhostbst2proteininteraction |