1887

Abstract

Ubiquinone is an essential redox component of the aerobic respiratory chains of bacteria and mitochondria. It is well established that mammalian ubiquinone can function in its reduced form (ubiquinol) as a lipid-soluble antioxidant preventing lipid peroxidation. The objective of this study was to test the hypothesis that prokaryotic ubiquinone is involved in the defence against oxidative stress in the cytoplasmic membrane. The rate of superoxide production by rapidly respiring wild-type membranes was twofold higher than in the slowly respiring membranes from a knockout mutant. However, large amounts of superoxide accumulated in the Ubi membranes compared to wild-type membranes, which possess superoxide-scavenging ubiquinol. Likewise, the rate of HO production was twofold higher in the wild-type, but the overall production of HO was again significantly higher in the Ubi membranes. Inclusion of a water-soluble ubiquinone homologue (UQ-1) effectively decreased the amount of HO produced in the Ubi membranes in a concentration-dependent manner. Addition of UQ-2 to the membranes was even more effective in limiting accumulation of HO than was UQ-1, suggesting a role for the side-chain in conferring liposolubility in the antioxidative defence mechanism. Intracellular HO concentration was increased 18-fold in the mutant, and expression of the gene, encoding the catalase hydroperoxidase I, as well as catalase enzyme activity, were increased twofold in this mutant. The mutant was hypersensitive to oxidative stress mediated by CuSO or HO; sensitivity to the latter could be abolished by addition of cysteine. This phenotype was also exhibited by a mutant, defective in the last step of UQ biosynthesis and therefore expected to accumulate several UQ biosynthetic intermediates. These observations support the participation of reduced ubiquinone as an antioxidant in . The mutant exhibited a pleiotropic phenotype, being resistant to heat, linolenic acid and phleomycin. Resistance to the two latter compounds is probably due to reduced uptake. Like mutants unable to synthesize the quinol oxidase, cytochrome , the mutant was also sensitive to dithiothreitol, an effect that is attributed to inability of the respiratory chain to maintain an appropriate redox balance in the periplasm.

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2000-04-01
2019-12-06
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References

  1. Afanas’ev, I. B., Korkina, L. G., Suslova, T. B. & Soodaeva, S. K. ( 1990; ). Are quinones producers or scavengers of superoxide ion in cells? Arch Biochem Biophys 281, 245-250.[CrossRef]
    [Google Scholar]
  2. Bader, M., Muse, W., Ballou, D. P., Gassner, C. & Bardwell, C. A. ( 1999; ). Oxidative protein folding is driven by the electron transport system. Cell 98, 217-227.[CrossRef]
    [Google Scholar]
  3. Beyer, R. E., Segura-Aguilar, J., Di Bernard, S. & 7 other authors ( 1996; ). The role of DT-diaphorase in the maintenance of the reduced antioxidant form of coenzyme Q in membrane systems. Proc Natl Acad Sci USA 93, 2528–2532.[CrossRef]
    [Google Scholar]
  4. Collis, C. M. & Grigg, G. W. ( 1989; ). An Escherichia coli mutant resistant to phleomycin, bleomycin and heat inactivation is defective in ubiquinone synthesis. J Bacteriol 171, 4792-4798.
    [Google Scholar]
  5. Compan, I. & Touati, D. ( 1993; ). Interaction of six global regulators in expression of manganese superoxide dismutase in Escherichia coli K-12. J Bacteriol 175, 1687-1696.
    [Google Scholar]
  6. Demple, B. ( 1991; ). Regulation of bacterial oxidative stress genes. Annu Rev Genet 25, 315-337.[CrossRef]
    [Google Scholar]
  7. D’mello, R., Hill, S. & Poole, R. K. ( 1996; ). The cytochrome bd quinol oxidase in Escherichia coli has an extremely high oxygen affinity and two oxygen-binding haems: implications for regulation of activity in vivo by oxygen inhibition. Microbiology 142, 755-763.[CrossRef]
    [Google Scholar]
  8. Do, T. Q., Schultz, J. R. & Clarke, C. ( 1996; ). Enhanced sensitivity of ubiquinone-deficient mutants of Saccharomyces cerevisiae to products of autoxidized polyunsaturated fatty acids. Proc Natl Acad Sci USA 93, 7534-7539.[CrossRef]
    [Google Scholar]
  9. Ernster, L. & Dallner, G. ( 1995; ). Biochemical, physiological and medical aspects of ubiquinone function. Biochim Biophys Acta 1271, 195-204.[CrossRef]
    [Google Scholar]
  10. Forsmark-Andrée, P., Dallner, G. & Ernster, L. ( 1995; ). Endogenous ubiquinol prevents protein modification accompanying lipid peroxidation in beef heart submitochondrial particles. Free Radic Biol Med 19, 749-757.[CrossRef]
    [Google Scholar]
  11. Gennis, R. B. & Stewart, V. (1996). Respiration. In Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd edn, pp. 217–261. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  12. Goldman, B. S., Gabbert, K. K. & Kranz, R. G. ( 1996a; ). Use of heme reporters for studies of cytochrome biosynthesis and heme transport. J Bacteriol 178, 6338-6347.
    [Google Scholar]
  13. Goldman, B. S., Gabbert, K. K. & Kranz, R. G. ( 1996b; ). The temperature-sensitive growth and survival phenotypes of Escherichia coli cydDC and cydAB strains are due to deficiencies in cytochrome bd and are corrected by exogenous catalase and reducing agents. J Bacteriol 178, 6348-6351.
    [Google Scholar]
  14. Gonzalez-Flecha, B. & Demple, B. ( 1994; ). Intracellular generation of superoxide as a by-product of Vibrio harveyi luciferase expressed in Escherichia coli. J Bacteriol 176, 2293-2299.
    [Google Scholar]
  15. Gonzalez-Flecha, B. & Demple, B. ( 1995; ). Metabolic sources of hydrogen peroxide in aerobically growing Escherichia coli. J Biol Chem 270, 13681-13687.[CrossRef]
    [Google Scholar]
  16. Hsu, A., Poon, W. W., Shepherd, J. A., Myles, D. C. & Clarke, C. ( 1996; ). Complementation of coq3 mutant yeast by mitochondrial targeting of the Escherichia coli UbiG polypeptide: evidence that UbiG catalyzes both O-methylation steps in ubiquinone biosynthesis. Biochemistry 35, 9797-9806.[CrossRef]
    [Google Scholar]
  17. Imlay, J. A. ( 1995; ). A metabolic enzyme that rapidly produces superoxide, fumarate reductase of Escherichia coli. J Biol Chem 270, 19767-19777.
    [Google Scholar]
  18. Imlay, J. A. & Fridovich, I. ( 1991; ). Assay of metabolic superoxide production in Escherichia coli. J Biol Chem 266, 6957-6965.
    [Google Scholar]
  19. Kalén, A., Norling, B., Appelkvist, E.-L. & Dallner, G. ( 1989; ). Ubiquinone biosynthesis by the microsomal fraction from rat liver. Biochim Biophys Acta 926, 70-78.
    [Google Scholar]
  20. Kimura, T. & Nishioka, H. ( 1997; ). Intracellular generation of superoxide by copper sulphate in Escherichia coli. Mutation Res 389, 237-242.[CrossRef]
    [Google Scholar]
  21. Kobayashi, T. & Ito, K. ( 1999; ). Respiratory chain strongly oxidizes the CXXC motif of DsbB in the Escherichia coli disulfide bond formation pathway. EMBO J 18, 1192-1198.[CrossRef]
    [Google Scholar]
  22. Kobayashi, T., Kishigama, S., Sone, M., Inokuchi, H., Mogi, T. & Ito, K. ( 1997; ). Respiratory chain is required to maintain oxidised states of the DsbA-DsbB disulfide bond formation system in aerobically growing Escherichia coli. Proc Natl Acad Sci USA 94, 11857-11862.[CrossRef]
    [Google Scholar]
  23. Loshen, G., Flohe, L. & Chance, B. ( 1971; ). Respiratory chain linked H2O2 production in pigeon heart mitochondria. FEBS Lett 18, 261-264.[CrossRef]
    [Google Scholar]
  24. Markwell, M. A. K., Haas, S. M., Bieber, L. L. & Tolbert, N. E. ( 1978; ). A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87, 206-210.[CrossRef]
    [Google Scholar]
  25. Messner, K. R. & Imlay, J. A. ( 1999; ). The identification of primary sites of superoxide and hydrogen peroxide formation in the aerobic respiratory chain and sulfite reductase complex of Escherichia coli. J Biol Chem 274, 10119-10128.[CrossRef]
    [Google Scholar]
  26. Missiakas, D. & Raina, S. ( 1997; ). Protein folding in the bacterial periplasm. J Bacteriol 179, 2465-2471.
    [Google Scholar]
  27. Nakayama, T., Hashimoto, M. & Hashimoto, K. ( 1997; ). Superoxide dismutase inhibition of oxidation of ubiquinol and concomitant formation of hydrogen peroxide. Biosci Biotechnol Biochem 61, 2034-2038.[CrossRef]
    [Google Scholar]
  28. Okada, K., Kainou, T., Matsuda, H. & Kawamukai, M. ( 1998; ). Biological significance of the side chain length in Saccharomyces cerevisiae. FEBS Lett 431, 241-244.[CrossRef]
    [Google Scholar]
  29. Olsson, J. M., Xia, L., Eriksson, L. C. & Bjornstedt, M. ( 1999; ). Ubiquinone is reduced by lipoamide dehydrogenase and this reaction is potently stimulated by zinc. FEBS Lett 448, 190-192.[CrossRef]
    [Google Scholar]
  30. Papa, S., Guerrieri, F. & Capitanio, N. ( 1997; ). A possible role of slips in cytochrome c oxidase in the antioxygen defense system of the cell. Biosci Rep 17, 23-31.[CrossRef]
    [Google Scholar]
  31. Poole, R. K. & Ingledew, W. J. (1987). Pathways of electrons to oxygen. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology, pp. 170–200. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  32. Poole, R. K., Anjum, M. F., Membrillo-Hernández, J., Kim, S. O., Hughes, M. N. & Stewart, V. ( 1996; ). Nitric oxide, nitrite, and Fnr regulation of hmp (flavohemoglobin) gene expression in Escherichia coli K-12. J Bacteriol 178, 5487-5492.
    [Google Scholar]
  33. Poon, W. W., Barkovich, R. J., Hsu, A. Y., Frankel, A., Lee, P. T., Shepherd, J. N., Myles, D. C. & Clarke, C. F. ( 1999; ). Yeast and rat Coq3 and Escherichia coli UbiG polypeptides catalyze both O-methyltransferase steps in coenzyme Q biosynthesis. J Biol Chem 274, 21665-21672.[CrossRef]
    [Google Scholar]
  34. Rosner, J. L. & Storz, G. ( 1997; ). Regulation of bacterial responses to oxidative stress. Curr Top Cell Regul 35, 163-175.
    [Google Scholar]
  35. Skulachev, V. P. ( 1997; ). Membrane-linked systems preventing superoxide formation. Biosci Rep 17, 347-366.[CrossRef]
    [Google Scholar]
  36. Søballe, B. & Poole, R. K. ( 1997; ). Aerobic and anaerobic regulation of the ubiCA operon, encoding the first two committed steps of ubiquinone biosynthesis in Escherichia coli. FEBS Lett 414, 373-376.[CrossRef]
    [Google Scholar]
  37. Søballe, B. & Poole, R. K. ( 1998; ). Requirement for ubiquinone downstream of cytochrome(s) b in the oxygen-terminated respiratory chains of Escherichia coli K-12 revealed using a null mutant allele of ubiCA. Microbiology 144, 361-373.[CrossRef]
    [Google Scholar]
  38. Søballe, B. & Poole, R. K. ( 1999; ). Microbial ubiquinones: multiple roles in respiration, gene regulation and oxidative stress management. Microbiology 145, 1817-1830.[CrossRef]
    [Google Scholar]
  39. Stroobant, P., Young, I. G. & Gibson, F. ( 1972; ). Mutants of Escherichia coli K-12 blocked in the final reaction of ubiquinone biosynthesis: characterization and genetic analysis. J Bacteriol 109, 134-139.
    [Google Scholar]
  40. Suzuki, K., Okada, K., Kamiya, Y., Zhu, X. F., Nakagawa, T., Kawamukai, M. & Matsuda, H. ( 1997; ). Analysis of the decaprenyl diphosphate synthase (dps) gene in fission yeast suggests a role of ubiquinone as an antioxidant. J Biochem 121, 496-505.[CrossRef]
    [Google Scholar]
  41. Thorn, J. M., Barton, J. D., Dixon, N. E., Ollis, D. L. & Edwards, K. J. ( 1995; ). Crystal structure of Escherichia coli QOR quinone oxidoreductase complexed with NADPH. J Mol Biol 249, 785-799.[CrossRef]
    [Google Scholar]
  42. Touati, D. ( 1988; ). Transcriptional and posttranscriptional regulation of manganese superoxide dismutase biosynthesis in Escherichia coli, studied with operon and protein fusions. J Bacteriol 170, 2511-2520.
    [Google Scholar]
  43. Wall, D., Delaney, J. M., Fayet, O., Lipinska, B., Yamamoto, T. & Georgopoulos, C. ( 1992; ). arc-dependent thermal regulation and extragenic suppression of the Escherichia coli cytochrome d operon. J Bacteriol 174, 6554-6562.
    [Google Scholar]
  44. Wallace, B. J. & Young, I. G. ( 1977; ). Role of quinones in electron transport to oxygen and nitrate in Escherichia coli: studies with a double quinone mutant. Biochim Biophys Acta 461, 84-100.[CrossRef]
    [Google Scholar]
  45. Wu, G., Williams, H. D., Zamanian, M., Gibson, F. & Poole, R. K. ( 1992; ). Isolation and characterization of Escherichia coli mutants affected in aerobic respiration: the cloning and nucleotide sequence of ubiG. Identification of an S-adenosylmethionine-binding motif in proteins, RNA, and small-molecule methyltransferases. J Gen Microbiol 138, 2101-2112.[CrossRef]
    [Google Scholar]
  46. Zeng, H., Snavely, P., Zamorano, P. & Javor, G. T. ( 1998; ). Low ubiquinone content in Escherichia coli causes thiol hypersensitivity. J Bacteriol 180, 3681-3685.
    [Google Scholar]
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