Potyvirus potato virus Y (PVY), which causes potato and other plants from the fattening family, has been infected shortly and was among the 10 most important
viruses from the economic and scientific point of view. During the cycle of infection of the coat proteins (CP) and other viral proteins, they react with each other
and with the host plant proteins, thereby ensuring optimal reproduction, encapsidation and movement of the virus. The plant also depends on these interactions
as it is part of its defence response. There are studies that describe the interaction of proteins within the Potyvirus-host system, however, most host proteins that
react with the unknown virus. A method that would help to explore the new interactions of proteins of the PVY-host system and at the same time allow research of
already existing interactions would greatly contribute to the field. The main aim of the study is to better understand protein interactions within the PVY-host
system. During the research we will select both viral and selected proteins host plants with a high-performance strategy, cloning them and looking at their
expression. The expressed proteins will be cleansed, and then with the help of Surface Plasma Resonance (SPR), we will induce the interactions between the
protein-protein system and the viral particle-protein. With the help of mutagenesis we will investigate the importance of specific amino acid residues and / or
protein motives during interaction. At the same time, SPRs will be used in plant extracts of various complexity to find proteins that react with non-mobile PVY
particles. The proteins that will bind to the PVY particles will be ejected from the SPR chip, and then identified by the use of mass spectrometry. In addition, we will
produce PVY-like particles and optimize their assembly. VLPs will be purified, used in SPR research, where they will be able to evaluate various CP mutations in the
interaction of various proteins with VLP. We will also try to obtain the 3D PVY CP structure, X-ray diffraction and electron microscopy. The acquired data, including
new interactions, additional kinetics and structural data, will be included in the signaling pathway for plant defense, which is currently in the dissemination phase.
In summary, the proposed study will greatly contribute to a better understanding of protein interactions that take place during PVY infection, reveal mechanisms
for the functioning of this important virus and allow for new strategies for the protection of plants against the virus. In addition, this research will contribute to the
field of new research tools such as SPR for measuring host-host interactions and PVY VLPs, which are promising tools that could be used for other biotechnology
industries such as epitopic carriers or presentations for immunological purposes.