Related Experiment Video
Updated: Jul 12, 2025
Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16
Published on: July 15, 2019
Inhibition of lysosome-tethered Ragulator-Rag-3D complex restricts the replication of Enterovirus 71 and Coxsackie
Xinhui Wang1, Zhilin Hu2, Wei Zhang1
1State Key Laboratory of Common Mechanism Research for Major Diseases, and Key Laboratory of Synthetic Biology Regulatory Elements, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Suzhou, China.
Insights
Researchers identified the Ragulator-Rag complex as crucial for Enterovirus 71 (EV71) and Coxsackie A16 (CVA16) replication and pathogenesis in hand, foot, and mouth disease (HFMD). A new inhibitor, ZHSI-1, targeting this complex shows promise for HFMD treatment.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Enterovirus 71 (EV71) and Coxsackie A16 (CVA16) are primary causes of hand, foot, and mouth disease (HFMD) in children.
- The precise mechanisms governing EV71 and CVA16 replication and pathogenesis are not fully elucidated.
Purpose of the Study:
- To identify host factors involved in EV71 and CVA16 replication and pathogenesis.
- To explore potential therapeutic targets for HFMD.
Main Methods:
- Genome-wide CRISPR-Cas9 knockout screening was employed to identify host factors.
- Viral protein interactions and cellular localization were investigated.
- A novel inhibitor, ZHSI-1, was synthesized and tested for antiviral activity.
- In vivo efficacy of ZHSI-1 was evaluated in mouse models of EV71 infection.
Main Results:
- The Ragulator-Rag complex was identified as essential for EV71-induced apoptosis and pyroptosis, and for EV71/CVA16 replication.
- The Ragulator-Rag complex recruits the viral 3D protein to lysosomes, facilitating viral replication.
- Disruption of the Ragulator-Rag-3D complex significantly inhibits viral replication.
- The novel inhibitor ZHSI-1 targets the 3D protein, preventing lysosomal tethering and repressing viral replication and pyroptosis.
- ZHSI-1 demonstrated protective effects against EV71 infection in mice.
Conclusions:
- The Ragulator-Rag complex plays a critical role in EV71 and CVA16 replication and pathogenesis by mediating lysosomal recruitment of viral proteins.
- Targeting the lysosome-tethered Ragulator-Rag-3D complex represents a promising therapeutic strategy for HFMD.
Abstract:
Enterovirus 71 (EV71) and Coxsackie A16 (CVA16) are two major causative agents of hand, foot, and mouth disease (HFMD) in young children. However, the mechanisms regulating the replication and pathogenesis of EV71/CVA16 remain incompletely understood. We performed a genome-wide CRISPR-Cas9 knockout screen and identified Ragulator as a mediator of EV71-induced apoptosis and pyroptosis. The Ragulator-Rag complex is required for EV71 and CVA16 replication. Upon infection, the Ragulator-Rag complex recruits viral 3D protein to the lysosomal surface through the interaction between 3D and RagB. Disruption of the lysosome-tethered Ragulator-Rag-3D complex significantly impairs the replication of EV71/CVA16. We discovered a novel EV71 inhibitor, ZHSI-1, which interacts with 3D and significantly reduces the lysosomal tethering of 3D. ZHSI-1 treatment significantly represses replication of EV71/CVA16 as well as virus-induced pyroptosis associated with viral pathogenesis. Importantly, ZHSI-1 treatment effectively protects against EV71 infection in neonatal and young mice. Thus, our study indicates that targeting lysosome-tethered Ragulator-Rag-3D may be an effective therapeutic strategy for HFMD.
Related Concept Videos
01:38Intralumenal Vesicles and Multivesicular Bodies
01:10Retrovirus Life Cycles