All respiring cells must solve the problem of oxidative stress, since respiration results in potentially toxic level of reactive oxygen species. Therefore, respiring cells contain the enzyme, superoxide dismutase, to catalyze the conversion of superoxide anion to hydrogen peroxide and molecular oxygen. Manganous superoxide dismutase (MnSOD) is found in the mitochondria of all multi-cellular eukaryotes, and is present in the yeast S. cerevisiae. The gene encoding MnSOD in yeast is called SOD1. Since the structural gene encoding MnSOD is well conserved and MnSOD enzymatic activity is essential for respiring cells, some features of SOD1 regulation in yeast are likely to be similar or identical to MnSOD gene regulation in more complex eukaryotes. Understanding SOD1 regulation, therefore, will contribute valuable information to the field of oxidative stress response. The investigator's objectives are to lay the ground work for understanding oxidative stress in yeast first, by finding mutants which specifically do not transcribe the SOD1 gene using an SOD1-lacZ fusion gene to identify these mutants, and standard genetic techniques to characterize them. The second objective is to complete a deletion analysis of the SOD1 promoter region of the SOD1-lacZ fusion gene by standard techniques. Included in the characterization of both the mutant strains and the promoter deletion will be a comparison of the expression of SOD1-lacZ in cells grown under anaerobic conditions, normal and higher than normal O2 conditions. This effort concerning the regulation of yeast manganous superoxide dismutase by oxygen and heme will broaden the knowledge of DNA binding proteins and their regulation and function. The project also furthers VPW program objectives to provide opportunities for women to advance their careers in science or engineering through research, and to encourage other women to pursue careers in these areas through the investigators' enhanced visibility as role models on the host campuses. In this project, the proposed activities which contribute to the second objective include: teaching a graduate level course entitled, Stress Responses in Eukaryotic Cells; organizing a workshop called, Getting There from Here about careers in biological science; and teaching a seminar course for undergraduate women in biological sciences entitled, Finding a Way: Women, Careers and Modern Biology.
