Jeroen Roelofs photoJeroen Roelofs
Associate Professor, Division of Biology
Kansas State University

Mechanism of chaperone-assisted assembly of proteasome regulatory particle

Mentor: Katsura Asano

The proteasome is the major cellular protease. It is involved in the controlled degradation of proteins that regulate a wide variety of cellular processes, such as transcription, apoptosis, cell divison and DNA repair. With an important role in homeostasis of so many proteins it is not surprising that observed increased proteasome activity (e.g. in multiple myelomas) or decreased proteasome activity (e.g. in many neurodegenerative diseases) is a pathological factor in many diseases. One determinant of cellular proteasome activity is the level of proteasomes in the cell. Thus, it is important to understand how cellular proteasome levels are regulated. The long term goal of this project is to understand how proteasome levels are regulated. Recent work has identified four chaperones that facilitate the formation of proteasome regulatory particle (RP). Each chaperone binds to the C-domain of a specific AAA-ATPases located in the RP. Despite this similarity in function and binding properties, there is no sequence or structural conservation among these chaperones.

The objective of this research is to understand the role each RP chaperone plays in the formation of proteasomes. We hypothesize that the chaperones act as templates for specific base subunits in the assembly of RP. Secondly, they regulate the order in which precursor complexes assemble in a spatial and temporal manner. We also hypothesize that the structural difference among the chaperones are important to accommodate as well as prevent a unique set of interactions for specific ATPases during the assembly process. We will use in vitro binding and reconstitution assays to study this. We furthermore will obtain structural information of the chaperones in combination with the C-domain they bind to. We expect that information from the expeiments proposed here will provided a detailed understanding of the mechanisms the chaperones employ to assist proteasome assembly. This could potentially provide new drug targets and ultimately enabling us to manipulate proteasome levels for therapeutic means.