Understanding Communications Between Viral RNAs and Human Proteins

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Background: Globally, viral infections are responsible for millions of deaths and impose a severe financial burden on healthcare systems. Viruses containing RNA as their genetic material have emerged as highly pathogenic and infectious. My interdisciplinary, integrative and innovative research program aims to provide critical insight into the human protein-viral RNA interactions that are of fundamental importance for viral replication. Aims and Approaches: The viral non-coding terminal regions are essential for virus replication. Therefore, over the next 5 years, my lab will investigate the structural features of 5' and 3' non-coding RNAs (Aim 1), identify human proteins that are being recruited by viral non-coding RNAs (Aim 2), and establish the structural basis of host protein - viral RNA complexes (Aim 3). Viral RNAs will be prepared using in vitro RNA transcription, and size exclusion chromatography. Using pull-down and mass-spectrometry assays, human proteins that specifically interact with viral RNAs will be identified. Human proteins will be expressed in E. coli, animal, or insect cells, followed by their purification by means of affinity and size-exclusion chromatography. Microscale thermophoresis will aid the determination of affinity between viral RNAs and human proteins. The viral RNA-human protein complexes will be characterized using light scattering and analytical ultracentrifuge techniques, followed by structural studies using solution X-ray scattering, and cryo-electron microscopy. Impact: The proposed research will highlight mechanisms by which viral RNAs exploit human proteins to aid viral replication. The identification and comparison of the structural features of viral non-coding RNAs could provide insights into how viral non-coding RNAs adapt and evolve, which can affect viral pathogenicity, spread, and resistance and influence agriculture, and biotechnology areas. Ultimately, the detailed insights of structural features of a broad range of RNA viruses can promote the urgently required framework to develop strategies to treat viral diseases by targeting host proteins - viral RNA complexes. HQP: My priority is to work on interdisciplinary research programs involving cutting-edge biochemical, biophysical, -omics, and structural biology techniques to create a high-quality, cross-institutional training environment to stimulate and foster the development of highly skilled research personnel. Such skills will help trainees with their future careers in academia, government agencies, and industries related to agriculture, biomedicine, or biotechnology research, and ultimately improve the quality of life of Canadians.