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Published on: January 26, 2019
A Proteomic-Based Workflow Using Purified Respiratory Syncytial Virus Particles to Identify Cellular Factors as Drug
Tra Nguyen Huong1, Boon Huan Tan2, Richard J Sugrue3
1School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, Singapore, 637551, Singapore.
Abstract:
The identification of cellular factors that play a role in respiratory syncytial virus (RSV) replication is an alternative strategy in the identification of druggable cellular protein that are essential for RSV replication. In this regard experimental strategies that are able to screen relevant proteins from the vast array of proteins in the cellular milieu will facilitate the identification of potential drug targets. In this chapter we describe a procedure where RSV particles are purified from cells that are permissive for RSV infection, and the protein composition of the purified virus particles characterized using a proteomics-based strategy. This procedure revealed that actin, several actin-binding proteins, and the chaperones HSP70 and HSP90 also co-purified with the virus particles. The relevance of the HSP90 protein to virus replication was then further validated using imaging, gene silencing and by using an established small molecule HSP90 inhibitor.
Insights
Identifying host cell proteins is key to finding new drugs against respiratory syncytial virus (RSV). This study found that HSP90 is crucial for RSV replication, suggesting it as a potential drug target.
Area of Science:
- Virology
- Proteomics
- Drug Discovery
Background:
- Respiratory syncytial virus (RSV) replication relies on host cell factors.
- Identifying essential cellular proteins can reveal druggable targets for antiviral therapies.
Purpose of the Study:
- To identify cellular proteins associated with purified RSV particles using proteomics.
- To investigate the role of identified host factors, particularly HSP90, in RSV replication.
Main Methods:
- Purification of RSV particles from infected cells.
- Proteomic analysis to characterize the protein composition of purified RSV.
- Validation of HSP90's role using imaging, gene silencing, and small molecule inhibitors.
Main Results:
- Proteomic analysis identified actin, actin-binding proteins, HSP70, and HSP90 co-purifying with RSV particles.
- HSP90 was confirmed to be relevant for RSV replication through experimental validation.
Conclusions:
- Host cell chaperones, specifically HSP90, are associated with purified RSV particles.
- HSP90 is a critical factor for RSV replication and represents a potential therapeutic target.
