A comprehensive analysis of essential protein regulators of clathrin mediated en...
A comprehensive analysis of essential protein regulators of clathrin mediated endocytic pathway
Clathrin-mediated endocytosis is an essential molecular process involved in the maintenance of cell homeostasis of all eukaryotic organisms. It is indispensable for the uptake of nutrients and signaling molecules, for the proper c...
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Descripción del proyecto
Clathrin-mediated endocytosis is an essential molecular process involved in the maintenance of cell homeostasis of all eukaryotic organisms. It is indispensable for the uptake of nutrients and signaling molecules, for the proper composition of plasma membrane, and for the regulation of multiple signaling pathways. Since endocytosis is a primary entry point to the cell interior, its detailed knowledge is of key importance for many biomedical disciplines like immunology, neurophysiology and pharmacology. Studies on endocytosis at neuronal synapses or on its role during viral infection are just two examples of current research linked to potential therapeutic applications. Clathrin-mediated endocytosis is an elaborate, highly dynamic process assisted by more than 60 proteins, the majority of which is evolutionarily conserved. Different multiprotein modules are involved in the initial sorting of endocytic cargo, formation of an endocytic vesicle, and its subsequent scission and internalization. Although many components are known, molecular mechanism and spatiotemporal order of endocytic events are only being to be understood. Moreover, the key endocytic factors are essential, multifunctional proteins of modular structure, which greatly complicates their analysis in human. We plan to study the role of essential endocytic genes by combining quantitative real-time fluorescence microscopy with the powerful genetics of yeast Saccharomyces cerevisie. Crucial endocytic proteins will be depleted from the cell and endocytosis will be followed by live-cell imaging. Complementation assays with truncated/mutated constructs will be used to delineate the molecular details of missing function. Electron microscopy techniques will be introduced to reveal structures of obtained endocytic intermediates. The focus on critical endocytic factors studied by innovative multidisciplinary approaches should bring a significant progress in our understanding of this highly orchestrated process.