Location: Foreign Animal Disease Research
Project Number: 3022-32000-063-020-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jul 1, 2020
End Date: Jun 30, 2024
Objective:
African Swine Fever (ASF) is a large DNA virus of approximately 180kb, and contains approximately 160 different open reading frames (ORFs) encoding for an ASFV protein, with little or no function. The primary target cell for ASFV in swine is macrophages. To identify a known function for these genes proteins one methodology is to identify the host proteins that interact with the various ASFV ORFs proteins. However only a select few of these proteins have had any attempts at protein crystallization. Protein crystallization allows a visual representation of the molecular structure of the protein. This research project seeks to determine the crystal structure and host-viral protein complexes involved in ASFV infection.
Approach:
Computer situ docking prediction of African Swine Fever viral protein-protein interactions will be initially completed. This docking allows for the identification of either critical residues that mediate this interaction, or to allow for the design of novel inhibitors. The next stage of the project will be to confirm that these protein-protein interactions occur at the molecular level under cellular conditions, in order to do this a Tap-tag mass septectomy procedure will be performed. Using extensive bioinformatics, from the mass spectrometry data and the in situ computer docking, we will then be able to evaluate a rational approach for disrupting these protein interactions either by mutating residues in the viral protein or creating an artificial blocking peptide. Using this data we will test to confirm that the predictions of critical residues mediating viral-host protein interactions are in fact correct under swine cellular conditions. Finally, we will evaluate the results of the prediction studies to the protein interactions that occur in our ASFV swine model system.
