Analysis of Candidate Proteins Interacting with Eyal

Abstract

The Eyal gene is important for mammalian organogenesis, and mutations in the human EYA1 results in Branchio-Oto-Renal (BOR) syndrome. This syndrome is autosomal dominant and causes combinations of branchial, otic, and renal anomalies. All Eya gene products contain an extremely conserved 271 amino acid C-terminus that is essential in protein-protein interaction. Recently, it was found that during mouse development, the expression of certain Six genes relied on the function of Eyal, and that Eyal interacts with relevant Six proteins through its C-terminal domain. These results provided insight into the molecular and developmental basis for organ defects occurring in BOR syndrome. Previous studies also suggested that there are direct and indirect interactions with Eyal and other proteins. Several candidate Eyal-interacting proteins have been recently isolated using yeast two-hybrid screens in P-X. Xu’s lab (McLaughlin Research Institute, Great Falls, Montana). Since the yeast two-hybrid screen is a sensitive system for protein-protein interaction, I sought to test the interaction by assaying five mutations in the Eyal domain on protein-protein interactions in yeast cells. I demonstrated preliminary evidence that four proteins specifically interact with Eyal. Further studies will be performed to test my findings.