Overproduction of the rbo gene product from Desulfovibrio species suppresses all deleterious effects of lack of superoxide dismutase in Escherichia coli.
In an attempt to isolate the superoxide dismutase (SOD) gene from the anaerobic sulfate-reducing bacterium Desulfoarculus baarsii, a DNA fragment was isolated which functionally complemented an Escherichia coli mutant (sodA sodB) deficient in cytoplasmic SODs. This region carries two open reading frames with sequences which are very similar to that of the rbo-rub operon from Desulfovibrio vulgaris. Independent expression of the rbo and rub genes from ptac showed that expression of rbo was responsible for the observed phenotype. rbo overexpression suppressed all deleterious effects of SOD deficiency in E. coli, including inactivation by superoxide of enzymes containing 4Fe-4S clusters and DNA damage produced via the superoxide-enhanced Fenton reaction. Thus, rbo restored to the sodA sodB mutant the ability to grow on minimal medium without the addition of branched amino acids, and growth on gluconate and succinate carbon sources was no longer impaired. The spontaneous mutation rate, which is elevated in SOD-deficient mutants, returned to the wild-type level in the presence of Rbo, which also restored aerobic viability of sodA sodB recA mutants. Rbo from Desulfovibrio vulgaris, but not Desulfovibrio gigas desulforedoxin, which corresponds to the NH2-terminal domain of Rbo, complemented sod mutants. The physiological role of Rbo in sulfate-reducing bacteria is unknown. In E. coli, Rbo may permit the bacterium to avoid superoxide stress by maintaining functional (reduced) superoxide sensitive 4Fe-4S clusters. It would thereby restore enzyme activities and prevent the release of iron that occurs after cluster degradation and presumably leads to DNA damage.
Pianzzola
M J
MJ
Facultad de Quimica, Catedra de Microbiologia, Montevideo, Uruguay.
Soubes
M
M
Touati
D
D
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D003901:Desulfovibrio / Q000235:genetics*; D016969:Desulfovibrio vulgaris / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D015870:Gene Expression; D005816:Genetic Complementation Test; D005942:Gluconates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D012416:Rubredoxins / Q000235:genetics / Q000378:metabolism; D013386:Succinates / Q000378:metabolism; D019802:Succinic Acid; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism
D003470:Culture Media; D005942:Gluconates; D012416:Rubredoxins; D013386:Succinates; D013481:Superoxides; D019802:Succinic Acid; D013482:Superoxide Dismutase; C023091:rubredoxin-NAD+ reductase; D009247:NADH, NADPH Oxidoreductases; D005947:Glucose; C030691:gluconic acid