Abstract 2433 Investigating the protein-protein interactions of Drosophila UNC-45

From fruit flies to humans, UNC-45 has demonstrated that it plays an important role in the proper development, differentiation, and repair processes of different cells and tissue types. For example, missense mutations and the aberrant expression of UNC-45 have resulted in increased cancer proliferation, disorganized muscle assembly, and encumbered intestinal epithelia development. With a structure and sequence that lacks any indication of performing catalytic functions, it's hypothesized that UNC-45 serves as a scaffold by recruiting other proteins to perform a specific function - mediating protein-protein interactions. Through the use of in vivo chemical crosslinking we have been able to capture transient and low-affinity protein-protein interactions of UNC-45 that occur throughout the pupae stage of Drosophila melanogaster, a period of intense development and differentiation for the fruit fly. Chemically crosslinked protein-complexes including UNC-45 have been purified and soon to be analyzed via high-resolution mass spectrometry to identify the proteins that interact with UNC-45 in vivo throughout development and differentiation. In conjunction with the in vivo experiments, we are in the process of developing a workflow of elucidating the interactive faces of known protein-protein interactions involving UNC-45 through the use of transmission electron microscopy and interferometric scattering microscopy. In developing the workflow, we are focused on studying the interaction between UNC-45 and HSP-83, and UNC-45 and tubulin protein. This research was funded in part by the California Metabolic Research Foundation.