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Decoding a virus Achilles heel: the African swine fever interactome (ASFVInt)
Project
Project code: 2821ERA26D
Contract period: 01.04.2021
- 31.03.2024
Budget: 116,958 Euro
Purpose of research: Experimental development
Keywords: diagnostics, prevention, pigs, animal health, virology, knowledge transfer, networking, zoonosis
The main goals of ASFVInt are the mapping of physical interactions between the proteins of the African swine fever virus (ASFV) and proteins of the host (pig) and the elucidation of so far unknown functions of ASFV proteins, creating a basis for the understanding of ASFV pathogenesis and identifying intervention points for antiviral strategies. As for the majority of ASFV genes neither functions nor interactors have been described so far, ASFVInt will implement two ‚open view‘ screening strategies (Yeast-Two-Hybrid [Y2H] and, at the FLI, affinity purification-mass spectrometry [AP-MS]) to identify the ASFV interactome. Identified protein-protein interactions (PPI) will be validated by the FLI partner with independent assays in order to select for the most promising PPI, which will be characterized in greater detail by biochemical assays and, finally, by cell culture experiments for functional characterisation. Biochemical studies will provide physico-chemical parameters of the interactions and identify binding domains, allowing the development of antiviral peptides and other small molecules. For the functional characterisation of PPI, ASFV deletion mutatns will be generated for future pathogenesis studies in the animal (not part of ASFVInt). Apart from these two deliverables, ASFVInt will provide a detailled interactome map which is based on experimentally validated data and will also include known intractions of the host proteins. FLI will provide two technologies which are critical for the success of the project, ‘affinity-purification-mass spectrometry’ (AP-MS) as screening tool fo new PPI, and ‚microscale thermophoresis‘ (MST), to determine the physico-chemical parameters of protein binding.
Section overview
Subjects
- Animal health
Funding programme
Excutive institution
Friedrich-Loeffler-Institute - Federal Research Institute for Animal Health (FLI)