1887

Abstract

Alternative oxidases (Aox or Aod) are present in the mitochondria of plants, fungi and many types of yeast. These enzymes transfer electrons from the ubiquinol pool directly to oxygen without contributing to the proton transfer across the mitochondrial membrane. Alternative oxidases are involved in stress responses, programmed cell death and maintenance of the cellular redox balance. The alternative oxidase gene of the methylotrophic yeast was isolated and cloned to study its regulation and the effects of deregulation of the alternative respiration by overexpression or disruption of the gene. Both disruption and overexpression had negative effects on the biomass yield; however, the growth rate and substrate uptake rate of the strain overexpressing the alternative oxidase were slightly increased. These effects were even more pronounced when higher glucose concentrations were used. The occurrence of free intracellular radicals and cell death phenomena was investigated using dihydrorhodamine 123 and the TUNEL test. The results suggest a major contribution of the alternative oxidase to cell viability. The negative effects of deregulated alternative respiration clearly indicated the importance of precise regulation of the alternative oxidase in this yeast.

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2007-04-01
2019-10-19
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References

  1. Affourtit, C., Albury, M. S., Krab, K. & Moore, A. L. ( 1999; ). Functional expression of the plant alternative oxidase affects growth of the yeast Schizosaccharomyces pombe. J Biol Chem 274, 6212–6218.[CrossRef]
    [Google Scholar]
  2. Affourtit, C., Albury, M. S., Crichton, P. G. & Moore, A. L. ( 2002; ). Exploring the molecular nature of alternative oxidase regulation and catalysis. FEBS Lett 510, 121–126.[CrossRef]
    [Google Scholar]
  3. Andersson, M. E. & Nordlund, P. ( 1999; ). A revised model of the active site of alternative oxidase. FEBS Lett 449, 17–22.[CrossRef]
    [Google Scholar]
  4. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidmann, J. G., Smith, J. A. & Struhl, K. ( 2003; ). Current Protocols in Molecular Biology. New York: Wiley.
  5. Berthold, D. A. & Siedow, J. N. ( 1993; ). Partial purification of the cyanide-resistant alternative oxidase of skunk cabbage (Symplocarpus foetidus) mitochondria. Plant Physiol 101, 113–119.[CrossRef]
    [Google Scholar]
  6. Berthold, D. A., Andersson, M. E. & Nordlund, P. ( 2000; ). New insight into the structure and function of the alternative oxidase. Biochim Biophys Acta 1460, 241–254.[CrossRef]
    [Google Scholar]
  7. Borghouts, C., Werner, A., Elthon, T. & Osiewacz, H. D. ( 2001; ). Copper-modulated gene expression and senescence in the filamentous fungus Podospora anserina. Mol Cell Biol 21, 390–399.[CrossRef]
    [Google Scholar]
  8. Borghouts, C., Scheckhuber, C. Q., Stephan, O. & Osiewacz, H. D. ( 2002a; ). Copper homeostasis and aging in the fungal model system Podospora anserina: differential expression of PaCtr3 encoding a copper transporter. Int J Biochem Cell Biol 34, 1355–1371.[CrossRef]
    [Google Scholar]
  9. Borghouts, C., Scheckhuber, C. Q., Werner, A. & Osiewacz, H. D. ( 2002b; ). Respiration, copper availability and SOD activity in P. anserina strains with different lifespan. Biogerontology 3, 143–153.[CrossRef]
    [Google Scholar]
  10. Chaudhuri, M., Ajayi, W., Temple, S. & Hill, G. C. ( 1995; ). Identification and partial purification of a stage-specific 33 kDa mitochondrial protein as the alternative oxidase of the Trypanosoma brucei brucei bloodstream trypomastigotes. J Eukaryot Microbiol 42, 467–472.[CrossRef]
    [Google Scholar]
  11. Claros, M. G. & Vincens, P. ( 1996; ). Computational method to predict mitochondrially imported proteins and their targeting sequences. Eur J Biochem 241, 779–786.[CrossRef]
    [Google Scholar]
  12. Crameri, A., Whitehorn, E. A., Tate, E. & Stemmer, W. P. ( 1996; ). Improved green fluorescent protein by molecular evolution using DNA shuffling. Nat Biotechnol 14, 315–319.[CrossRef]
    [Google Scholar]
  13. Dufour, E., Boulay, J., Rincheval, V. & Sainsard-Chanet, A. ( 2000; ). A causal link between respiration and senescence in Podospora anserina. Proc Natl Acad Sci U S A 97, 4138–4143.[CrossRef]
    [Google Scholar]
  14. Fröhlich, K. U. & Madeo, F. ( 2003; ). Genetics of Apoptosis: Regulators and Applications of Yeast Apoptosis. Oxford: Oxford University Press.
  15. Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M. R., Appel, R. D. & Bairoch, A. ( 2005; ). Protein identification and analysis tools on the ExPASy server. In Proteomics Protocols Handbook, pp. 571–607. Edited by J. M. Walker. Totowa, NJ: Humana Press.
  16. Gonzalez-Barroso, M. M., Ledesma, A., Lepper, S., Perez-Magan, E., Zaragoza, P. & Rial, E. ( 2006; ). Isolation and bioenergetic characterization of mitochondria from Pichia pastoris. Yeast 23, 307–313.[CrossRef]
    [Google Scholar]
  17. Gonzalez-Meler, M. A., Ribas-Carbo, M., Giles, L. & Siedow, J. N. ( 1999; ). The effect of growth and measurement temperature on the activity of the alternative respiratory pathway. Plant Physiol 120, 765–772.[CrossRef]
    [Google Scholar]
  18. Huang, X., von Rad, U. & Durner, J. ( 2002; ). Nitric oxide induces transcriptional activation of the nitric oxide-tolerant alternative oxidase in Arabidopsis suspension cells. Planta 215, 914–923.[CrossRef]
    [Google Scholar]
  19. Huh, W. K. & Kang, S. O. ( 2001; ). Characterization of the gene family encoding alternative oxidase from Candida albicans. Biochem J 356, 595–604.[CrossRef]
    [Google Scholar]
  20. Huq, S. & Palmer, J. M. ( 1978; ). Isolation of a cyanide-resistant duroquinol oxidase from Arum maculatum mitochondria. FEBS Lett 95, 217–220.[CrossRef]
    [Google Scholar]
  21. Johnson, C. H., Prigge, J. T., Warren, A. D. & McEwen, J. E. ( 2003; ). Characterization of an alternative oxidase activity of Histoplasma capsulatum. Yeast 20, 381–388.[CrossRef]
    [Google Scholar]
  22. Joseph-Horne, T., Babij, J., Wood, P. M., Hollomon, D. & Sessions, R. B. ( 2000; ). New sequence data enable modelling of the fungal alternative oxidase and explain an absence of regulation by pyruvate. FEBS Lett 481, 141–146.[CrossRef]
    [Google Scholar]
  23. Joseph-Horne, T., Hollomon, D. W. & Wood, P. M. ( 2001; ). Fungal respiration: a fusion of standard and alternative components. Biochim Biophys Acta 1504, 179–195.[CrossRef]
    [Google Scholar]
  24. Kirimura, K., Matsui, T., Sugano, S. & Usami, S. ( 1996; ). Enhancement and repression of cyanide-insensitive respiration in Aspergillus niger. FEMS Microbiol Lett 141, 251–254.[CrossRef]
    [Google Scholar]
  25. Kita, K., Nihei, C. & Tomitsuka, E. ( 2003; ). Parasite mitochondria as drug target: diversity and dynamic changes during the life cycle. Curr Med Chem 10, 2535–2548.[CrossRef]
    [Google Scholar]
  26. Lorin, S., Dufour, E., Boulay, J., Begel, O., Marsy, S. & Sainsard-Chanet, A. ( 2001; ). Overexpression of the alternative oxidase restores senescence and fertility in a long-lived respiration-deficient mutant of Podospora anserina. Mol Microbiol 42, 1259–1267.
    [Google Scholar]
  27. Lorin, S., Dufour, E. & Sainsard-Chanet, A. ( 2006; ). Mitochondrial metabolism and aging in the filamentous fungus Podospora anserina. Biochim Biophys Acta 1757, 604–610.[CrossRef]
    [Google Scholar]
  28. Madeo, F., Frohlich, E. & Frohlich, K. U. ( 1997; ). A yeast mutant showing diagnostic markers of early and late apoptosis. J Cell Biol 139, 729–734.[CrossRef]
    [Google Scholar]
  29. Madeo, F., Frohlich, E., Ligr, M., Grey, M., Sigrist, S. J., Wolf, D. H. & Frohlich, K. U. ( 1999; ). Oxygen stress: a regulator of apoptosis in yeast. J Cell Biol 145, 757–767.[CrossRef]
    [Google Scholar]
  30. Maxwell, D. P., Wang, Y. & McIntosh, L. ( 1999; ). The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells. Proc Natl Acad Sci U S A 96, 8271–8276.[CrossRef]
    [Google Scholar]
  31. Meeuse, B. J. D. ( 1975; ). Thermogenic respiration in aroids. Annu Rev Plant Physiol 26, 117–126.[CrossRef]
    [Google Scholar]
  32. Moore, A. L., Albury, M. S., Crichton, P. G. & Affourtit, C. ( 2002; ). Function of the alternative oxidase: is it still a scavenger? Trends Plant Sci 7, 478–481.[CrossRef]
    [Google Scholar]
  33. Nihei, C., Fukai, Y., Kawai, K., Osanai, A., Yabu, Y., Suzuki, T., Ohta, N., Minagawa, N., Nagai, K. & Kita, K. ( 2003; ). Purification of active recombinant trypanosome alternative oxidase. FEBS Lett 538, 35–40.[CrossRef]
    [Google Scholar]
  34. Ordog, S. H., Higgins, V. J. & Vanlerberghe, G. C. ( 2002; ). Mitochondrial alternative oxidase is not a critical component of plant viral resistance but may play a role in the hypersensitive response. Plant Physiol 129, 1858–1865.[CrossRef]
    [Google Scholar]
  35. Osiewacz, H. D. ( 2002; ). Mitochondrial functions and aging. Gene 286, 65–71.[CrossRef]
    [Google Scholar]
  36. Osiewacz, H. D. & Stumpferl, S. W. ( 2001; ). Metabolism and aging in the filamentous fungus Podospora anserina. Arch Gerontol Geriatr 32, 185–197.[CrossRef]
    [Google Scholar]
  37. Robson, C. A. & Vanlerberghe, G. C. ( 2002; ). Transgenic plant cells lacking mitochondrial alternative oxidase have increased susceptibility to mitochondria-dependent and -independent pathways of programmed cell death. Plant Physiol 129, 1908–1920.[CrossRef]
    [Google Scholar]
  38. Sainsard-Chanet, A. & Begel, O. ( 1990; ). Insertion of an LrDNA gene fragment and of filler DNA at a mitochondrial exon-intron junction in Podospora. Nucleic Acids Res 18, 779–783.[CrossRef]
    [Google Scholar]
  39. Sakajo, S., Minagawa, N. & Yoshimoto, A. ( 1998; ). Regulation of alternative oxidase gene expression by carbon source in the yeast Hansenula anomala. In Plant Mitochondria: from Gene to Function, Proceedings of the International Congress, Aronsborg, Sweden, June 22–27, 1998, pp. 515–518.
  40. Schulte, E., Kuck, U. & Esser, K. ( 1989; ). Multipartite structure of mitochondrial DNA in a fungal longlife mutant. Plasmid 21, 79–84.[CrossRef]
    [Google Scholar]
  41. Shi, N. Q., Davis, B., Sherman, F., Cruz, J. & Jeffries, T. W. ( 1999; ). Disruption of the cytochrome c gene in xylose-utilizing yeast Pichia stipitis leads to higher ethanol production. Yeast 15, 1021–1030.[CrossRef]
    [Google Scholar]
  42. Shi, N. Q., Cruz, J., Sherman, F. & Jeffries, T. W. ( 2002; ). SHAM-sensitive alternative respiration in the xylose-metabolizing yeast Pichia stipitis. Yeast 19, 1203–1220.[CrossRef]
    [Google Scholar]
  43. Siedow, J. N. & Umbach, A. L. ( 2000; ). The mitochondrial cyanide-resistant oxidase: structural conservation amid regulatory diversity. Biochim Biophys Acta 1459, 432–439.[CrossRef]
    [Google Scholar]
  44. Siedow, J. N., Umbach, A. L. & Moore, A. L. ( 1995; ). The active site of the cyanide-resistant oxidase from plant mitochondria contains a binuclear iron center. FEBS Lett 362, 10–14.[CrossRef]
    [Google Scholar]
  45. Simons, B. H., Millenaar, F. F., Mulder, L., Van Loon, L. C. & Lambers, H. ( 1999; ). Enhanced expression and activation of the alternative oxidase during infection of Arabidopsis with Pseudomonas syringae pv tomato. Plant Physiol 120, 529–538.[CrossRef]
    [Google Scholar]
  46. Tanton, L. L., Nargang, C. E., Kessler, K. E., Li, Q. & Nargang, F. E. ( 2003; ). Alternative oxidase expression in Neurospora crassa. Fungal Genet Biol 39, 176–190.[CrossRef]
    [Google Scholar]
  47. Thompson, J. D., Higgins, D. G. & Gibson, T. J. ( 1994; ). clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673–4680.[CrossRef]
    [Google Scholar]
  48. Tsuda, A., Witola, W. H., Ohashi, K. & Onuma, M. ( 2005; ). Expression of alternative oxidase inhibits programmed cell death-like phenomenon in bloodstream form of Trypanosoma brucei rhodesiense. Parasitol Int 54, 243–251.[CrossRef]
    [Google Scholar]
  49. Tudella, V. G., Curti, C., Soriani, F. M., Santos, A. C. & Uyemura, S. A. ( 2004; ). In situ evidence of an alternative oxidase and an uncoupling protein in the respiratory chain of Aspergillus fumigatus. Int J Biochem Cell Biol 36, 162–172.[CrossRef]
    [Google Scholar]
  50. Umbach, A. L. & Siedow, J. N. ( 2000; ). The cyanide-resistant alternative oxidases from the fungi Pichia stipitis and Neurospora crassa are monomeric and lack regulatory features of the plant enzyme. Arch Biochem Biophys 378, 234–245.[CrossRef]
    [Google Scholar]
  51. Van Hellemond, J. J., Simons, B., Millenaar, F. F. & Tielens, A. G. ( 1998; ). A gene encoding the plant-like alternative oxidase is present in Phytomonas but absent in Leishmania spp. J Eukaryot Microbiol 45, 426–430.[CrossRef]
    [Google Scholar]
  52. Vanlerberghe, G. C., Robson, C. A. & Yip, J. Y. ( 2002; ). Induction of mitochondrial alternative oxidase in response to a cell signal pathway down-regulating the cytochrome pathway prevents programmed cell death. Plant Physiol 129, 1829–1842.[CrossRef]
    [Google Scholar]
  53. Veiga, A., Arrabaca, J. D. & Loureiro-Dias, M. C. ( 2000; ). Cyanide-resistant respiration is frequent, but confined to yeasts incapable of aerobic fermentation. FEMS Microbiol Lett 190, 93–97.[CrossRef]
    [Google Scholar]
  54. Veiga, A., Arrabaca, J. D. & Loureiro-Dias, M. C. ( 2003a; ). Cyanide-resistant respiration, a very frequent metabolic pathway in yeasts. FEMS Yeast Res 3, 239–245.[CrossRef]
    [Google Scholar]
  55. Veiga, A., Arrabaca, J. D. & Loureiro-Dias, M. C. ( 2003b; ). Stress situations induce cyanide-resistant respiration in spoilage yeasts. J Appl Microbiol 95, 364–371.[CrossRef]
    [Google Scholar]
  56. Veiga, A., Arrabaca, J. D., Sansonetty, F., Ludovico, P., Corte-Real, M. & Loureiro-Dias, M. C. ( 2003c; ). Energy conversion coupled to cyanide-resistant respiration in the yeasts Pichia membranifaciens and Debaryomyces hansenii. FEMS Yeast Res 3, 141–148.[CrossRef]
    [Google Scholar]
  57. Weis, R., Luiten, R., Skranc, W., Schwab, H., Wubbolts, M. & Glieder, A. ( 2004; ). Reliable high-throughput screening with Pichia pastoris by limiting yeast cell death phenomena. FEMS Yeast Res 5, 179–189.[CrossRef]
    [Google Scholar]
  58. Yabu, Y., Yoshida, A., Suzuki, T., Nihei, C., Kawai, K., Minagawa, N., Hosokawa, T., Nagai, K., Kita, K. & Ohta, N. ( 2003; ). The efficacy of ascofuranone in a consecutive treatment on Trypanosoma brucei brucei in mice. Parasitol Int 52, 155–164.[CrossRef]
    [Google Scholar]
  59. Zaslaver, A., Mayo, A. E., Rosenberg, R., Bashkin, P., Sberro, H., Tsalyuk, M., Surette, M. G. & Alon, U. ( 2004; ). Just-in-time transcription program in metabolic pathways. Nat Genet 36, 486–491.[CrossRef]
    [Google Scholar]
  60. Zottini, M., Formentin, E., Scattolin, M., Carimi, F., Lo Schiavo, F. & Terzi, M. ( 2002; ). Nitric oxide affects plant mitochondrial functionality in vivo. FEBS Lett 515, 75–78.[CrossRef]
    [Google Scholar]
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