New Genetic Tools for Comparative Analysis of Emerging Viruses and Virus-Host Molecular Interactions in Reservoir Hosts versus Spillover Hosts
Technical Report,01 Apr 2018,30 Mar 2019
Trustees of Boston College Chestnut Hill United States
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Emerging viruses pose significant problems for military personnel living in close quarters, andor deployed overseas. Significant gaps remain in our understanding of the underlying molecular mechanisms of viral emergence. To fill this gap in knowledge we seek to develop a robust method for identifying the binding partners of small viral proteins implicated in modifying host cell defenses. We are employing a cutting edge genetic system for incorporating synthetic amino-acids at defined positions in proteins within cellular systems these non-canonical amino-acids ncAAs are modified with a side chain that can be induced by UV light to covalently cross-link cellular binding partners. This effectively tags the target proteins allowing subsequent identification by massspec. We have generated the necessary mutants of two viral proteins, Vpr of HIV-1and ORF4a of MERS-Coronavirus, and have conducted a pilot mass-spec experiment. All tools and reagents are now in place, and in the nextfinal period of this 18-monthDiscovery Award we plan to move forward with mass-spec identification and experimental confirmation of candidate binding partners.
- Medicine and Medical Research