Isolation and Characterization of the sod2$^{2+}$ Gene Encoding a Putative Mitochondrial Manganese Superoxide Dismutase in Schizosaccharomyces bombe

  • Jeong, Jae-Hoon (Laboratory of Molecular Microbiology, School of Biological Sciences, and Institute of Microbiology, Seoul National University) ;
  • Kwon, Eun-Soo (Laboratory of Molecular Microbiology, School of Biological Sciences, and Institute of Microbiology, Seoul National University) ;
  • Roe, Jung-Hye (Laboratory of Molecular Microbiology, School of Biological Sciences, and Institute of Microbiology, Seoul National University)
  • Published : 2001.03.01

Abstract

The fission yeast Schizosaccharomyces pombe contains two distinct superoxide dismutase (SOD) activities, one in the cytosol encoded by the $sod2^{+}$ gene and the other in mitochondria. The $sod2^{+}$ gene encoding putative mitochondrial manganese superoxide dismutase (MnSOD) was isolated from the S. pombe genomic library using a PCR fragment as the probe. The nucleotide sequence of the $sod2^{+}$ gene and its flanking region (4051 bp HindIII fragment) was determined. An intron of 123 nt in size was predicted and confirmed by sequencing the cDNA following reverse transcription PCR. The predicted Sod2p consists of 218 amino acid residues with a molecular mass of 24,346 Da. The deduced amino acid sequence showed a high degree of homology with other MnSODs, especially in the metal binding residues at the active site and their relative positions. The transcriptional start site was mapped by primer extension at 231 at upstream from the ATG codon. A putative TATA box(TATAAAA) was located 58 nt upstream from the transcriptional start site and putative polyadenylation sites were located at 1000, 1062, and 1074 nt downstream from the ATG start codon.

Keywords

References

  1. CRC Crit. Rev. Biochem. v.22 Aspects of the structure, function, and applications of superoxide dismutase Bannister J.V.;W.H. Bannister;G. Rotilio
  2. Prog. Nucleic Acid Res. Mol. Biol. v.40 Superoxide dismutases Beyer W.;J. Imlay;I. Fridovich
  3. Annu. Rev. biochem. v.64 Superoxide radical and superoxide dismutases Fridovich I.
  4. Mol. Gen. Genet. v.215 Genetic engineering of Schizosaccharomyces pombe: A system for gene disruption and replacement using the ura4 gene as a selectable marker Grimm C.;J. Kohli;J. Murray;K. Maundrell
  5. Yeast v.14 Oxidative stress responses of the yeast Saccharomyces cerevisiae Jamieson D.J.
  6. J. Biol. Chem. v.245 Superoxide dismutase from Escherichia coli B.;A new manganese-containing enzyme Keele B.B. Jr.;J.M. McCord;I. Fridovich
  7. Eur. J. Biochem. v.241 Differential expression of superoxide dismutases containing Ni and Fe/Zn in Streptomyces coelicolor Kim E.J.;H.P. Kim;Y.C. Hah;J.H. Roe
  8. Microbiology v.141 Adaptive response of Schizosaccharomyces pombe to hydrogen peroxide and menadione Lee J.;I.W. Dawes;J.H. Roe
  9. J. Microbiol. v.38 Subcellular localization of catalase encoded by the ctt1+ gene in Schizosaccharomyces pombe Lee S.;J. Lee;J.H. Roe
  10. J. Biol. Chem. v.244 Superoxide dismutase;An enzymic function for erythrocuprein (hemocuprein) McCord J.M.;I. Fridovich
  11. Methods Enzymol. v.194 Molecular genetic analysis of fission yeast Schizosaccharomyces pombe Moreno S.;A. Klar;P. Nurse
  12. Molecular Biology of the Fission Yeast Nasim A.;P. Young;B.F. Johnson
  13. Annu. Rev. Biochem. v.59 The mitochondrial protein import apparatus Pfanner N.;W. Neupert
  14. Molecular Cloning: A Laboratory Manual Sambrook J.;E.F. Fritsch;T. Maniatis
  15. Proc. Natl. Acad. Sci. USA v.91 Oxidative damage and mitochondrial decay in aging Shigenaga M.K.;T.M. Hagen;B.N. Ames
  16. Genes Dev. v.12 SAPKs and transcription factors do the nucleocytoplasmic tango Wilkinson M.G.;J.B.A. Millar
  17. J. Biol. Chem. v.248 An iron-containing superoxide dismutase from Escherichia coli Yost F.J.;I. Fridovich
  18. Biochem. J. v.318 A novel nickel-containing superoxide dismutase from streptomyces spp. Youn H.D.;E.J. Kim;J.H. Roe;Y.C. Hah;S.O. Kang
  19. DNA. Cell. Biol. v.14 Schizosaccharomyces pombe: a model for molecular studies of eukaryotic genes Zhao Y.;H.B. Lieberman