Super-resolution optical microscopy studies of peroxisomal protein import in the yeast Saccharomyces cerevisiae

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Peroxisomes are single-membrane-bound organelles, which are ubiquitous in nearly all eukaryotic cells. The importance of peroxisomes is highlighted by a group of very severe metabolic diseases, which are caused by mutations in the peroxisomal biogenesis factors. Progress has been made in understanding the basic principles of peroxisomal functions using biochemical tools. However, techniques such as super-resolution optical STED (Stimulated Emission Depletion) microscopy are required to directly study dynamic processes in living cells. Initiated by the DFG Research Unit 1905 (Structure and function of the peroxisomal translocon), a collaboration between the groups of Prof. Erdmann at the Ruhr University of Bochum and Prof. Eggeling at the University of Oxford has been established, combining the long-standing expertise of both groups on peroxisomes and microscopy. Successful STED microscopy investigations on peroxisomes were performed in human cells. For a better understanding of the function of the individual peroxisomal proteins we need to precisely modify them genetically for defects in peroxisome biogenesis, as realized in yeast strains from the Erdmann lab. The aim is to apply STED microscopy to study the dynamic organization of proteins involved in peroxisomal protein import in yeast. First staining and handling protocols will be established as well as data acquisition and analysis will be optimized, which will lay the basis for using the available yeast strains to gain a better understanding of the basic mechanisms of peroxisomal protein import and proliferation.