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J Bacteriol. Dec 1996; 178(23): 6736–6742.
PMCID: PMC178569

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.

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Selected References

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  • Balch WE, Fox GE, Magrum LJ, Woese CR, Wolfe RS. Methanogens: reevaluation of a unique biological group. Microbiol Rev. 1979 Jun;43(2):260–296. [PMC free article] [PubMed]
  • Beauchamp C, Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Brumlik MJ, Voordouw G. Analysis of the transcriptional unit encoding the genes for rubredoxin (rub) and a putative rubredoxin oxidoreductase (rbo) in Desulfovibrio vulgaris Hildenborough. J Bacteriol. 1989 Sep;171(9):4996–5004. [PMC free article] [PubMed]
  • Brumlik MJ, Leroy G, Bruschi M, Voordouw G. The nucleotide sequence of the Desulfovibrio gigas desulforedoxin gene indicates that the Desulfovibrio vulgaris rbo gene originated from a gene fusion event. J Bacteriol. 1990 Dec;172(12):7289–7292. [PMC free article] [PubMed]
  • Canfield DE, Des Marais DJ. Aerobic sulfate reduction in microbial mats. Science. 1991 Mar 22;251:1471–1473. [PubMed]
  • Carlioz A, Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. [PMC free article] [PubMed]
  • Chen L, Liu MY, Legall J, Fareleira P, Santos H, Xavier AV. Purification and characterization of an NADH-rubredoxin oxidoreductase involved in the utilization of oxygen by Desulfovibrio gigas. Eur J Biochem. 1993 Sep 1;216(2):443–448. [PubMed]
  • Chen L, Liu MY, LeGall J, Fareleira P, Santos H, Xavier AV. Rubredoxin oxidase, a new flavo-hemo-protein, is the site of oxygen reduction to water by the "strict anaerobe" Desulfovibrio gigas. Biochem Biophys Res Commun. 1993 May 28;193(1):100–105. [PubMed]
  • Farr SB, D'Ari R, Touati D. Oxygen-dependent mutagenesis in Escherichia coli lacking superoxide dismutase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8268–8272. [PMC free article] [PubMed]
  • Flint DH, Emptage MH, Guest JR. Fumarase a from Escherichia coli: purification and characterization as an iron-sulfur cluster containing enzyme. Biochemistry. 1992 Oct 27;31(42):10331–10337. [PubMed]
  • Flint DH, Tuminello JF, Emptage MH. The inactivation of Fe-S cluster containing hydro-lyases by superoxide. J Biol Chem. 1993 Oct 25;268(30):22369–22376. [PubMed]
  • Fürste JP, Pansegrau W, Frank R, Blöcker H, Scholz P, Bagdasarian M, Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. [PubMed]
  • Gardner PR, Fridovich I. Superoxide sensitivity of the Escherichia coli 6-phosphogluconate dehydratase. J Biol Chem. 1991 Jan 25;266(3):1478–1483. [PubMed]
  • Gardner PR, Fridovich I. Superoxide sensitivity of the Escherichia coli aconitase. J Biol Chem. 1991 Oct 15;266(29):19328–19333. [PubMed]
  • Gregory EM, Moore WE, Holdeman LV. Superoxide dismutase in anaerobes: survey. Appl Environ Microbiol. 1978 May;35(5):988–991. [PMC free article] [PubMed]
  • Hatchikian EC, Henry YA. An iron-containing superoxide dismutase from the strict anaerobe Desulfovibrio desulfuricans (Norway 4). Biochimie. 1977;59(2):153–161. [PubMed]
  • Hewitt J, Morris JG. Superoxide dismutase in some obligately anaerobic bacteria. FEBS Lett. 1975 Feb 15;50(3):315–318. [PubMed]
  • Imlay JA, Fridovich I. Assay of metabolic superoxide production in Escherichia coli. J Biol Chem. 1991 Apr 15;266(11):6957–6965. [PubMed]
  • Keyer K, Gort AS, Imlay JA. Superoxide and the production of oxidative DNA damage. J Bacteriol. 1995 Dec;177(23):6782–6790. [PMC free article] [PubMed]
  • Kuo CF, Mashino T, Fridovich I. alpha, beta-Dihydroxyisovalerate dehydratase. A superoxide-sensitive enzyme. J Biol Chem. 1987 Apr 5;262(10):4724–4727. [PubMed]
  • Liochev SI, Fridovich I. Fumarase C, the stable fumarase of Escherichia coli, is controlled by the soxRS regulon. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5892–5896. [PMC free article] [PubMed]
  • Liochev SI, Fridovich I. The role of O2.- in the production of HO.: in vitro and in vivo. Free Radic Biol Med. 1994 Jan;16(1):29–33. [PubMed]
  • Moura I, Tavares P, Moura JJ, Ravi N, Huynh BH, Liu MY, LeGall J. Purification and characterization of desulfoferrodoxin. A novel protein from Desulfovibrio desulfuricans (ATCC 27774) and from Desulfovibrio vulgaris (strain Hildenborough) that contains a distorted rubredoxin center and a mononuclear ferrous center. J Biol Chem. 1990 Dec 15;265(35):21596–21602. [PubMed]
  • Nakayama K. Rapid viability loss on exposure to air in a superoxide dismutase-deficient mutant of Porphyromonas gingivalis. J Bacteriol. 1994 Apr;176(7):1939–1943. [PMC free article] [PubMed]
  • Natvig DO, Imlay K, Touati D, Hallewell RA. Human copper-zinc superoxide dismutase complements superoxide dismutase-deficient Escherichia coli mutants. J Biol Chem. 1987 Oct 25;262(30):14697–14701. [PubMed]
  • Rouault TA, Klausner RD. Iron-sulfur clusters as biosensors of oxidants and iron. Trends Biochem Sci. 1996 May;21(5):174–177. [PubMed]
  • Sancar A, Hack AM, Rupp WD. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Santos H, Fareleira P, Xavier AV, Chen L, Liu MY, LeGall J. Aerobic metabolism of carbon reserves by the "obligate anaerobe" Desulfovibrio gigas. Biochem Biophys Res Commun. 1993 Sep 15;195(2):551–557. [PubMed]
  • Tavares P, Ravi N, Moura JJ, LeGall J, Huang YH, Crouse BR, Johnson MK, Huynh BH, Moura I. Spectroscopic properties of desulfoferrodoxin from Desulfovibrio desulfuricans (ATCC 27774). J Biol Chem. 1994 Apr 8;269(14):10504–10510. [PubMed]
  • Touati D. Cloning and mapping of the manganese superoxide dismutase gene (sodA) of Escherichia coli K-12. J Bacteriol. 1983 Sep;155(3):1078–1087. [PMC free article] [PubMed]
  • Touati D, Jacques M, Tardat B, Bouchard L, Despied S. Lethal oxidative damage and mutagenesis are generated by iron in delta fur mutants of Escherichia coli: protective role of superoxide dismutase. J Bacteriol. 1995 May;177(9):2305–2314. [PMC free article] [PubMed]

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