Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae

Journal Article · Thu Mar 01 00:00:00 EST 1990 · Journal of Bacteriology; (USA)

OSTI ID:6897852

Bowler, C; Inze, D; Van Camp, W; Kaer, L V; Dhaese, P ^[1]

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1. Rijksuniversiteit Gent (Belgium)

Recombinant clones containing the manganese superoxide dismutase (MnSOD) gene of Bacillus stearothermophilus were isolated with an oligonucleotide probe designed to match a part of the previously determined amino acid sequence. Complementation analyses, performed by introducing each plasmid into a superoxide dismutase-deficient mutant of Escherichia coli, allowed us to define the region of DNA which encodes the MnSOD structural gene and to identify a promoter region immediately upstream from the gene. These data were subsequently confirmed by DNA sequencing. Since MnSOD is normally restricted to the mitochondria in eucaryotes, we were interested (i) in determining whether B. stearothermophilus MnSOD could function in eucaryotic cytosol and (ii) in determining whether MnSOD could replace the structurally unrelated copper/zinc superoxide dismutase (Cu/ZnSOD) which is normally found there. To test this, the sequence encoding bacterial MnSOD was cloned into a yeast expression vector and subsequently introduced into a Cu/ZnSOD-deficient mutant of the yeast Saccharomyces cerevisiae. Functional expression of the protein was demonstrated, and complementation tests revealed that the protein was able to provide tolerance at wild-type levels to conditions which are normally restrictive for this mutant. Thus, in spite of the evolutionary unrelatedness of these two enzymes, Cu/ZnSOD can be functionally replaced by MnSOD in yeast cytosol.

• OSTI ID: 6897852
• Journal Information: Journal of Bacteriology; (USA), Vol. 172:3; ISSN 0021-9193
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
SUPEROXIDE DISMUTASE
GENES
BACILLUS
COPPER
DNA SEQUENCING
ESCHERICHIA COLI
MANGANESE
MUTANTS
PLASMIDS
RECOMBINANT DNA
SACCHAROMYCES CEREVISIAE
ZINC
BACTERIA
CELL CONSTITUENTS
DNA
ELEMENTS
ENZYMES
EUMYCOTA
FUNGI
METALS
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PLANTS
SACCHAROMYCES
STRUCTURAL CHEMICAL ANALYSIS
TRANSITION ELEMENTS
YEASTS
550200* - Biochemistry