Research objectives and content
Plectin is a widely expressed protein of very large size that has all the attributes of a multifunctional crosslinking and organizing element of the cytoskeleton. It displays a multi-domain structure, versatile binding activities, and subcellular localizations that enable it to strengthen cells against mechanical stress forces. Biochemical studies have shown that plectin's interactions are differentially regulated by protein kinases, one of these being the cell-cycle control kinase p34cdc2 which phosphorylates plectin at a unique site during mitosis. During this phase plectin's ability to bind to vimentin in vitro decreases and it's solubility increases. Apart from this site, plectin's tail region contains a large number of potential phosphoacceptors, some of which occur in the recently found vimentin binding site. This project intends to investigate the role that phosphorylation in plectin's tail domain has on its interactions and functions, by studying interactions of bacterially expressed plectin mutant proteins (PMP's) and synthetic peptides in various phosphorylation states with intermediate filaments using binding assays, ultrastructural studies, and transfection studies.
Training content (objective, benefit and expected impact)
The proposed project will constitute a doctoral thesis project at the University of Vienna. The results of this project will contribute to the knowledge of plectin's functions and interactions, and its role in maintenance of cellular architecture. The aim is to report the results of this project to international journals, as well as writing a doctoral thesis on them. My expectation is that being involved in this project will not only greatly improve my experience with the methods and techniques mentioned, but will also develop the scientific abilities and skills necessary to work as a researcher.
Links with industry / industrial relevance (22)