Structural and biochemical studies elucidating the biogenesis pathway of 20S proteasomes

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In this proposal we address the spatial and temporal order of events in the chaperone-guided assembly and maturation of proteasomal 20S core particle (CP) complexes. We aim to structurally characterise the late steps of CP biogenesis by determining high-resolution cryo EM reconstructions in order to identify the exact interactions between proteasomal chaperones and 20S CP subunits. At the same time, we want to analyse by which mechanism(s) autocatalytic activation of the beta subunits is initiated. In parallel, we want to investigate the so far poorly understood early steps of proteasome CP biogenesis biochemically and structurally through the characterization of individual assembly intermediates, which will moreover be used to recapitulate distinct assembly steps in vitro with the aim to determine the order of events and their checkpoints.