Location: Foreign Animal Disease Research
Project Number: 8064-32000-060-23-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jun 1, 2019
End Date: May 31, 2024
The objective of this collaborative research project is to continue the assessment of the role of specific Classical Swine Fever Virus (CSFV) proteins in the process of virus replication and virulence. It is expected knowledge obtained here may enable the development of novel countermeasurement to control virus replication.
CSFV proteins and derived peptides will be assessed for their insertion, oligomeric assembly and pore formation in model membranes. These models comprise: i) lipid monolayers (measurements of lateral pressure in Langmuir troughs); ii) unilamellar vesicles with defined diameters (LUV) prepared according to the extrusion method through the measurement of water- membrane partitioning, lipid-mixing and aqueous contents release through spectrofluorimetric methods, iii) Giant Unilamellar Vesicles prepared according to the electroformation method to establish pore size through confocal microscopy analyses, and iv) Suspended planar phospholipid bilayers to evaluate ion channel activity by electrophysiological measurements. These complementary approaches will be used to define sequence and lipid requirements for the function of the CSFV proteins. Insertion into membranes implies acquisition of defined secondary structure. Thus, circular dichroism and infrared spectroscopy measurements will be additionally carried out to follow membrane-partitioning coupled structuring. Identification and describe the molecular basis of the interaction between viral proteins interacting with membranes in the host cells will contribute to understand mechanism of virus replication and disease production may be used for developing novel tools for controlling virus infection in vivo.