This research project will focus on using molecular techniques to characterize a new family of proteins (the Ebp2 proteins). The Ebp2 proteins are significant because they are required for eukaryotic cellular division. Since the molecular and genetic techniques available in yeast provide powerful approaches for studying basic aspects of cellular biology, the EBP2 homolog from S. cerevisiae will be studied. Preliminary analysis indicates that yeast Ebp2p is an essential nucleolar protein, and that ebp2 mutants suffer from defects related to rRNA metabolism and chromosome segregation. In order to gain a greater understanding of the role that Ebp2p plays in cellular metabolism, new ebp2 mutants will be created and characterized. Random and site-directed mutagenesis will be used to alter critical residues within the Ebp2 protein, and these mutations will be studied to determine the effects that they have on nucleolar metabolism and nuclear division. The genetic approaches of suppressor analysis and synthetic lethality will be used to identify proteins that interact with Ebp2p. To further define the role that Ebp2p plays in nuclear metabolism, microscopic analysis will be used to probe nuclear structure in wild type and mutant ebp2 cells. The Ebp2 protein will also be investigated to determine if it is associated with chromatin and whether it functions in chromosome condensation of the nucleolar rDNA repeat sequences. Since the analysis of a novel protein in yeast (Ebp2p) offers a great opportunity to engage students in basic research, these studies will be coordinated with the education of students in molecular genetics. Also, in collaboration with the Project to Increase Mastery of Mathematics and Science (PIMMS), a Biotechnology Workshop will be developed to train high school and middle school teachers in experiments in basic molecular biology and genetics. These workshops will use yeast and bacteria as model organisms, and they will emphasize hands-on approaches for teaching basic concepts of genetics to high school and middle school students.
