Intellectual Merit: Proper replication and maintenance of the DNA genome is essential for cell growth, function, and viability. When cells are exposed to environmental stresses that damage DNA, pathways for repairing the DNA damage are induced. One key player in this damage control is Replication Protein A (RPA), a complex of three proteins that serves as a "bridge" between damaged DNA and the machinery needed to repair the damage. In this capacity, RPA has the potential to interact with many proteins, but how specificity of those interactions is determined is not clear. One idea is that stress-induced phosphorylation of RPA mediates differential interaction between RPA and various proteins involved in DNA repair. This research will test that hypothesis by identifying which amino acid residues in the RPA proteins are phosphorylated, how phosphorylation affects physical interactions between RPA and different protein partners, and what impact RPA phosphorylation has on the ability of cells to maintain proper growth and viability. The research will be carried out in baker's yeast, which is amenable to both genetic and biochemical manipulation and is accessible to researchers of all skill levels. Because RPA is highly conserved (from yeast to human), the research findings will advance understanding of how eukaryotes coordinate DNA repair processes for maintaining genome integrity and cellular viability.
Broader Impacts: Centered at North Dakota State University, this project will contribute to learning and research opportunities science for college students in rural North Dakota and surrounding states by: improving molecular biology course content; expanding educational opportunities for non-traditional students through development of distance-learning courses in molecular biology, cell biology, genetics and biochemistry; providing opportunities for undergraduate students to conduct hands-on laboratory research; and providing a pipeline for interested students to obtain master's degrees as prerequisite for entering a new doctoral research program in STEM education. By fostering student literacy in science, this project will enable North Dakota State University to produce graduates with a solid scientific foundation that could positively impact their future contributions to society.
