NSF Award
9973746
In this POWRE grant, the principal investigator, who is at a small college, will spend a sabbatical year of research at the University of North Carolina at Chapel Hill in the laboratory of Dr. Sharon L.Milgram, where she will learn techniques of cell and molecular biology that can be applied to problems in cell signaling in a variety of model systems. The research will specifically test the hypothesis that protein kinase A (PKA) is anchored at the apical cell surface by A-kinase anchoring proteins (AKAPs) and that this anchoring is required for PKA-mediated regulation of apical ion transport. In Aim 1, the PI will learn cell culture techniques and establish stable epithelial cell lines expressing the soluble HT31 peptide fused with the greeen fluorescent protein (GFP). HT31 is known to bind PKA with high affinity, and removes PKA from membranes when overexpressed in cells. The PI will test the hypothesis that efficient modulation<br/>of apical ion channels requires membrane-associate PKA. In Aim 2 the PI will learn a variety of molecular biology techniques to clone and characterize AKAPs expressed in airway epithelial cells. Site-directed mutagenesis and expression in cell lines will be done to identify PKA binding sites in the cloned AKAPs, and to determine whether novel AKAPs target the cell surface in epithelia. In addition the PI will explore the function of<br/>epithelial AKAPs using cell culture model systems, and will determine whether<br/>particular AKAPs are required for efficient modulation of apical ion transport. <br/>A secondary goal of this POWRE application is to develop research<br/>projects that can be performed by undergraduates at Fort Lewis College, and to establish a long-term relationship between students at Fort Lewis College and faculty at the University of North Carolina. Since Fort Lewis College has an enrollment of ~12 % Native Americans, the proposal also impacts on the education of minority students
