Targeting the host factor HGS–viral membrane protein interaction in coronavirus infection
Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li
Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li
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Research Article Cell biology Microbiology Virology

Targeting the host factor HGS–viral membrane protein interaction in coronavirus infection

Abstract

While current antivirals primarily target viral proteins, host-directed strategies remain underexplored. Here, we performed a genome-wide CRISPR inhibition (CRISPRi) screening to identify the host protein, hepatocyte growth factor-regulated tyrosine kinase substrate (HGS), facilitating the pan-coronavirus infection both in vitro and in vivo. Mechanistically, HGS interacts with the viral membrane (M) protein, facilitating its trafficking to the ER-Golgi intermediate compartment for virion assembly. Conversely, HGS deficiency caused M retention in the ER, blocking assembly. Leveraging this interaction, we designed M-derived peptides and screened over 5,000 FDA-approved or commonly used drugs, identifying riboflavin tetrabutyrate (RTB). Both the peptides and RTB bind HGS and disrupt its interaction with the M protein, leading to M retention in the ER and subsequent blockade of virion assembly. These agents demonstrated broad anti-pan-coronavirus activity in vitro and in vivo. Collectively, our findings establish HGS as a druggable host target and identify RTB as a promising broad-spectrum antiviral candidate.

Authors

Xubing Long, Rongrong Chen, Rong Bai, Buyun Tian, Yu Cao, Kangying Chen, Fuyu Li, Yiliang Wang, Yongjie Tang, Qi Yang, Liping Ma, Fan Wang, Maoge Zhou, Xianjie Qiu, Yongzhi Lu, Jie Zheng, Peng Zhou, Xinwen Chen, Qian Liu, Xuepeng Wei, Yongxia Shi, Yanhong Xue, Jincun Zhao, Wei Ji, Liqiao Hu, Jinsai Shang, Tao Xu, Zonghong Li

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Figure 3

HGS interacts with M protein and facilitates its trafficking to ERGIC.

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HGS interacts with M protein and facilitates its trafficking to ERGIC. (...
(A and B) Representative IF analysis of the colocalization of SARS-CoV-2 M with SEC61B or ERGIC53 in WT and Hgs-KO 17Cl-1 cells (A) or in SARS-CoV2 Omicron BA.5-infected Huh7.5.1 WT and Hgs-KO cells (MOI = 0.5) (B). Trace outline is used for line-scan analysis of the relative fluorescence intensity of SARS-CoV-2 M with SEC61B and ERGIC53. Scale bar, 10 μm. Quantitative image analysis of M-SEC61B/ERGIC53 colocalization using Manders’ coefficient. N = 3 independent biological replications. (C) Representative IF analysis of the colocalization of HGS with SARS-CoV-2 M, S, E, and N in Vero E6 cells. Trace outline is used for line-scan analysis of the relative fluorescence intensity of HGS with SARS-CoV-2 M, S, E, and N. Scale bar, 10 μm. N = 3 independent biological replications. (D–F) Co-IP analysis of interaction of individually transiently expressed SARS-CoV-2 M, S, E, N with HGS in HEK293T cells (D and E) and HGS with M, S, N in SARS-CoV-2–infected Vero-E6 cells (hpi = 24 h) (F). Immunoprecipitates pulled down by FLAG/HA (D) or HGS (E and F) antibody were analyzed by IB with indicated antibodies. IgG IP was used as negative control. Input represents 5% of the total cell extract. Molecular weights are in kDa. N = 3 independent biological replications. (G) In vitro pull-down analysis of the interaction between HGS and SARS-CoV-2 M. Purified HIS-HGS and FLAG-M proteins subjected to pull-down assay by FLAG antibody were analyzed by IB with indicated antibodies. FLAG-GST protein was used as a negative control. Input represents 5% of the total proteins used for pull-down. Molecular weights are in kDa. N = 3 independent biological replications. Data are the mean ± SD. Significance testing for A and B was performed with a 2-tailed t test. *P ≤ 0.05, ***P ≤ 0.0005.