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.
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Acknowledgments
We acknowledge the Singapore NRF and the SMART (ID program) for funding support Ms Anuradha Radhakrishnan for technical support and Dr Tang Kai and the proteomic core facility at SBS for assistance in running the protein digests in the 1D LC MS/MS machine.
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Huong, T.N., Tan, B.H., Sugrue, R.J. (2016). A Proteomic-Based Workflow Using Purified Respiratory Syncytial Virus Particles to Identify Cellular Factors as Drug Targets. In: Tripp, R., Jorquera, P. (eds) Human Respiratory Syncytial Virus. Methods in Molecular Biology, vol 1442. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3687-8_13
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DOI: https://doi.org/10.1007/978-1-4939-3687-8_13
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