New Genetic Tools for Comparative Analysis of Emerging Viruses and Virus-Host Molecular Interactions in Reservoir Hosts versus Spillover Hosts
Abstract
Emerging viruses pose significant problems for military personnel living in close quarters, and/or 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 next/final period of this 18-monthDiscovery Award we plan to move forward with mass-spec identification and experimental confirmation of candidate binding partners.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2019
- Accession Number
- AD1093912
Entities
People
- Welkin E Johnson
Organizations
- Boston College