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

Abstract

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 inmore » yeast cytosol.« less

Authors:
; ; ; ;  [1]
  1. (Rijksuniversiteit Gent (Belgium))
Publication Date:
OSTI Identifier:
6897852
Alternate Identifier(s):
OSTI ID: 6897852
Resource Type:
Journal Article
Journal Name:
Journal of Bacteriology; (USA)
Additional Journal Information:
Journal Volume: 172:3; Journal ID: ISSN 0021-9193
Country of Publication:
United States
Language:
English
Subject:
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

Citation Formats

Bowler, C., Inze, D., Van Camp, W., Kaer, L.V., and Dhaese, P. Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae. United States: N. p., 1990. Web.
Bowler, C., Inze, D., Van Camp, W., Kaer, L.V., & Dhaese, P. Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae. United States.
Bowler, C., Inze, D., Van Camp, W., Kaer, L.V., and Dhaese, P. Thu . "Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae". United States.
@article{osti_6897852,
title = {Characterization of the Bacillus stearothermophilus manganese superoxide dismutase gene and its ability to complement copper/zinc superoxide dismutase deficiency in Saccharomyces cerevisiae},
author = {Bowler, C. and Inze, D. and Van Camp, W. and Kaer, L.V. and Dhaese, P.},
abstractNote = {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.},
doi = {},
journal = {Journal of Bacteriology; (USA)},
issn = {0021-9193},
number = ,
volume = 172:3,
place = {United States},
year = {1990},
month = {3}
}