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Mechanisms and physiology of targeted protein degradation

Prof. Dr. Nico Dissmeyer, Dipl.-Biochem.

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We study post-translational modifications of proteins with an impact on protein homeostasis (proteostasis) and the biological relevance of the associated protein stabilities in the cell. We investigate the relevance of the different stabilities of proteins in cells and tissues of model plants. To this end, we trace molecular components of involved pathways and identify proteins that are specifically recognized and degraded. We have developed a genetic technique based on conditional proteolysis that allows proteins to be enriched or degraded as needed in living plants, insects and cells, thereby switching their function on and off.

Research topics

• identification of substrates of the protein-modifying and protein-degrading N-degron pathway.
• physiological relevance of N-degron-mediated proteolysis
• characterization of the protein-modifying enzymes of the N-degron pathway
• application of conditional controlled proteolysis versus accumulation in basic research and biotechnology
• Molecular Farming and Synthetic Biology in plants

Model systems

• in vivo: plants such as Arabidopsis thaliana, Australian tobacco (Nicotiana benthamiana) and barley (Hordeum vulgare), but also bacteria such as Escherichia coli and the soil bacterium Agrobacterium tumefaciens, which can genetically modify (transform) plants
• plant cell cultures (protoplasts) derived from various mutant plant lines
• in vitro, we use numerous approaches based on recombinant proteins, synthetische peptides or cell-free systems

Methods

• heterologous protein expression in bacteria, yeasts, insect cells, plants and their purification by different methods
• molecular genetic and physiological analyses of plants as well as standard methods, especially comparative plant performance experiments
• peptide-based analyses, peptide arrays and pulldowns
• assays for post-translational modifications and various protein degradation assays in vitro and in vivo
• fluorescence microscopy, fluorescence-based protein chemistry
• variety of plant-based assays, including stable genetic transformation of living plants and transient transformation of plant organs and cell cultures