Expression of the maize MnSod (Sod3) gene in MnSOD-deficient yeast rescues the mutant yeast under oxidative stress.
Superoxide dismutases (SOD) are ubiquitous in aerobic organisms and are believed to play a significant role in protecting cells against the toxic, often lethal, effect of oxygen free radicals. However, direct evidence that SOD does in fact participate in such a protective role is scant. The MnSOD-deficient yeast strain (Sod2d) offered an opportunity to test the functional role of one of several SOD isozymes from the higher plant maize in hopes of establishing a functional bioassay for other SODs. Herein, we present evidence that MnSOD functions to protect cells from oxidative stress and that this function is conserved between species. The maize Sod3 gene was introduced into the yeast strain Sod2d where it was properly expressed and its product processed into the yeast mitochondrial matrix and assembled into the functional homotetramer. Most significantly, expression of the maize Sod3 transgene in yeast rendered the transformed yeast cells resistant to paraquat-induced oxidative stress by complementing the MnSOD deficiency. Furthermore, analyses with various deletion mutants of the maize SOD-3 transit peptide in the MnSOD-deficient yeast strain indicate that the initial portion (about 8 amino acids) of the maize transit peptide is required to direct the protein into the yeast mitochondrial matrix in vivo to function properly. These findings indicate that the functional role of maize MnSOD is conserved and dependent on its proper subcellular location in the mitochondria of a heterologous system.
Zhu
D
D
Department of Genetics, North Carolina State University, Raleigh 27695-7614.
Scandalios
J G
JG
D005609:Free Radicals; D005816:Genetic Complementation Test; D007527:Isoenzymes / Q000235:genetics; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation; D010100:Oxygen / Q000378:metabolism; D010269:Paraquat / Q000633:toxicity; D010944:Plants / Q000201:enzymology / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics*; D003313:Zea mays / Q000201:enzymology / Q000235:genetics
D005609:Free Radicals; D007527:Isoenzymes; D013482:Superoxide Dismutase; D010269:Paraquat; D010100:Oxygen