Evidence for a novel role of copper-zinc superoxide dismutase in zinc metabolism

The LYS7 gene in Saccharomyces cerevisiae encodes a protein (yCCS) that del ivers copper to the active site of copper-zinc superoxide dismutase (CuZn-S OD, a product of the SOD) gene). In yeast lacking Lys7 (lys7 Delta), the SO D1 polypeptide is present but inactive. Mutants lacking the SOD1 polypeptid e (sod1 Delta) and lys7 Delta yeast show very similar phenotypes, namely po or growth in air and aerobic auxotrophies for lysine and methionine. Here, we demonstrate certain phenotypic differences between these strains: 1) lys 7 Delta cells are slightly less sensitive to paraquat than sod1 Delta cells , 2) EPR-detectable or "free" iron is dramatically elevated in sod1 Delta m utants but not in lys7 Delta yeast, and 3) although sod1 Delta mutants show increased sensitivity to extracellular zinc, the lys7 Delta strain is as r esistant as wild type. To restore the SOD catalytic activity but not the zi nc-binding capability of the SOD1 polypeptide, we overexpressed Mn-SOD from Bacillus stearothermophilus in the cytoplasm of sod1 Delta yeast. Paraquat resistance was restored to wild-type levels, but zinc was not. Conversely, expression of a mutant CuZn-SOD that binds zinc but has no SOD activity (H 46C) restored zinc resistance but not paraquat resistance. Taken together, these results strongly suggest that CuZn-SOD, in addition to its antioxidan t properties, plays a role in zinc homeostasis.