UROP Research Mentor Project Submission Portal: Submission #1272

Overall Research Project Description The 26S proteasome is an unusually large ATP-dependent self-compartmentalized protease responsible for most intracellular protein degradation in eukaryotes. As the terminal component of the ubiquitin proteasome system, it plays a pivotal role in regulating essential cellular processes, including cell cycle progression and gene expression. Formation of proteasomes requires the precise interactions of ~66 individual proteins that must rapidly assemble in the dense cellular milieu. Efforts to understand the assembly process have thus been inferred from static snapshots of presumed assembly intermediates, leading to the proposal of conflicting assembly pathways and uncertainty regarding the timing and kinetics of the process. To address this, we have adapted a non-radioactive pulse-chase system to dynamically track subunit incorporation and the localization of assembly events in living cells. The trainee work directly with the graduate research assistant to generate a comprehensive kinetic model of proteasome biogenesis, and help to elucidate WHERE in the cell these assembly steps occur. This innovative integrated and quantitative approach will provide new insights into assembly of multi-subunit complexes in general, and will inform efforts to prevent or reverse proteasome insufficiency in human diseases. Importantly, this knowledge can be impactful to a host of human diseases associated with dysregulated homeostasis, including cancer, musculoskeletal diseases, and neurodegenerative diseases.