1887

Abstract

The possible physiological role of mitochondria in anaerobically grown was investigated via enzyme localization and inhibitor studies. Almost all of the activity of citrate synthase (EC 4.1.3.7) was recovered in the mitochondrial fraction after differential centrifugation of spheroplast lysates. The enzyme exhibited a high degree of latency which was demonstrated by sonication of the mitochondrial fractions. Since citrate synthase is an important enzyme in anabolic reactions, a consequence of this localization is the requirement for transport of metabolites across the mitochondrial membranes. Such transport is likely to require energy which, as a result of anaerobiosis, cannot be supplied by respiration. It was therefore investigated whether ATP translocation into the mitochondria by an ADP/ATP translocase might be involved in anaerobic mitochondrial energy metabolism. It was shown that addition of the ADP/ATP translocase inhibitor bongkrekic acid to anaerobic cultures indeed inhibited growth, although only partially. It is concluded that mitochondria of fulfil a vital role in anaerobic sugar metabolism.

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1994-11-01
2021-07-29
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References

  1. Andreasen A.A., Stier T.J.B. Anaerobic nutrition of Saccharomyces cerevisiae. I. Ergosterol requirement for growth in a defined medium. J Cell Comp Physiol 1953; 41:23–26
    [Google Scholar]
  2. Andreasen A.A., Stier T.J.B. Anaerobic nutrition of S cerevisiae. II. Unsaturated fatty acid requirement for growth in a defined medium.. J Cell Comp Physiol 1954; 43:271–281
    [Google Scholar]
  3. Baker K.P., Schatz G. Mitochondrial proteins essential for viability mediate protein transport into yeast mitochondria. Nature 1991; 349:205–208
    [Google Scholar]
  4. Bruinenberg P.M., van Dijken J.P., Scheffers W.A. An enzymic analysis of NADPH production and consumption in Candida utilis CBS 621. J Gen Microbiol 1983; 129:965–971
    [Google Scholar]
  5. Bruinenberg P.M., van Dijken J.P., Kuenen J.G., Scheffers W.A. Critical parameters in the isolation of mitochondria from Candida utilis grown in continuous culture. J Gen Microbiol 1985; 131:1035–1042
    [Google Scholar]
  6. Cartledge T.G., Lloyd D. Subcellular fractionation by zonal centrifugation of glucose-repressed anaerobically grown Saccharomyces carlsbergensis. Biochem J 1972; 127:693–703
    [Google Scholar]
  7. Cartledge T.G., Lloyd D. Changes in enzyme activities and distributions during glucose derepression and respiratory adaptation of anaerobically grown Saccharomyces carlsbergensis. Biochem J 1973; 132:609–621
    [Google Scholar]
  8. Cartledge T.G., Lloyd D., Erecińska M., Chance B. The development of the respiratory chain of Saccharomyces carlsbergensis during respiratory adaptation. Biochem J 1972; 130:739–747
    [Google Scholar]
  9. Chapman C., Bartley W. The kinetics of enzyme changes in yeast under conditions that cause the loss of mitochondria. Biochem J 1968; 107:455–465
    [Google Scholar]
  10. Criddle R.S., Schatz G. Promitochondria of anaerobically grown yeast I. Isolation and biochemical properties.. Biochemistry 1969; 8:322–334
    [Google Scholar]
  11. Damsky C.H., Nelson W.M., Claude A. Mitochondria in anaerobically-grown, lipid-limited brewer’s yeast. J Cell Biol 1969; 43:174–179
    [Google Scholar]
  12. Eilers M., Oppliger W., Schatz G. Both ATP and an energized inner membrane are required to import a purified precursor protein into mitochondria. EMBO J 1987; 6:1073–1077
    [Google Scholar]
  13. Erdelt H., Weidemann M.J., Buchholz M., Klingenberg M. Some principal effects of bongkrekic acid on the binding of adenine nucleotides to mitochondrial membranes. Eur J Biochem 1972; 30:107–112
    [Google Scholar]
  14. Gawaz M., Douglas M.G., Klingenberg M.K. Structure-function studies of adenine nucleotide transport in mitochondria. II. Biochemical analysis of distinct AAC1 and AAC2 protein in yeast. J Biol Chem 1990; 265:14202–14208
    [Google Scholar]
  15. Gbelská Y., Šubík J., Svoboda A., Goffeau A., Kováč L. Intramitochondrial ATP and cell functions: yeast cells depleted of intramitochondrial ATP lose the ability to grow and multiply. Eur J Biochem 1983; 130:281–286
    [Google Scholar]
  16. Groot G.S.P., Kováč L., Schatz G. Promitochondria of anaerobically grown yeast. V. Energy transfer in the absence of an electron transfer chain. Proc Natl Acad Sci 1971; 68:308–311
    [Google Scholar]
  17. Harder W., Visser K., Kuenen J.G. Laboratory fermenter with an improved magnetic drive. Lab Pract 1974; 23:644–645
    [Google Scholar]
  18. Jauniaux J.-C., Urrestarazu L.A., Wiame J.-M. Arginine metabolism in Saccharomyces cerevisiae: subcellular localization of the enzymes. J Bacteriol 1978; 133:1096–1107
    [Google Scholar]
  19. Jenkins R.O., Cartledge T.G., Lloyd D. Respiratory adaptation of anaerobically grown Saccharomyces uvarum: changes in distribution of enzymes. J Gen Microbiol 1984; 130:2809–2816
    [Google Scholar]
  20. Jensen R.E., Yaffe M.P. Input of proteins into yeast mitochondria: the nuclear MAS2 gene encodes a component of the processing protease that is homologous to the MAS1-encoded subunit. EMBO J 1988; 7:3863–3871
    [Google Scholar]
  21. Klingenberg M. The ADP/ATP carrier in mitochondrial membranes. In The Enzymes of Biological Membranes 1985 Edited by Martonosi A.N. New York: Plenum Press; 4 pp 511–553
    [Google Scholar]
  22. Knirsch M., Gawaz P., Klingenberg M. The isolation and reconstitution of the ADP/ATP carrier from wild-type Saccharomyces cerevisiae. Identification of primarily one type (AAC-2). FEBS Lett 1989; 244:427–432
    [Google Scholar]
  23. Kolarov J., Kolarova N., Nelson N. A third ADP/ATP translocator gene in yeast. J Biol Chem 1990; 265:12711–12716
    [Google Scholar]
  24. Krämer R., Klingenberg M. Reconstitution of adenine nucleotide transport from beef heart mitochondria. Biochemistry 1979; 18:4209–4215
    [Google Scholar]
  25. Krämer R., Klingenberg M. Modulation of the reconstituted adenine nucleotide exchange by membrane potential. Biochemistry 1980; 19:556–560
    [Google Scholar]
  26. Lewin A.S., Hines V., Small G.M. Citrate synthase encoded by the CIT2 gene of Saccharomyces cerevisiae is peroxisomal. Mol Cell Biol 1990; 10:1399–1405
    [Google Scholar]
  27. Linnane A.W. Aspects of the biosynthesis of the mitochondria of Saccharomyces cerevisiae. In Oxidases and Related Redox Systems 1965 Edited by King T.E., Mason H.S., Morrison M. New York: John Wiley & Sons; pp 1102–1128
    [Google Scholar]
  28. Lowenstein J.M. The tricarboxylic acid cycle. In Metabolic Pathways 1967 Edited by Greenberg D.M. New York : Academic Press; 1 pp 146–254
    [Google Scholar]
  29. Pfanner N., Tropschug M., Neupert W. Mitochondrial protein import: nucleoside triphosphates are involved in conferring import-competence to precursors. Cell 1987; 49:19–27
    [Google Scholar]
  30. Rogers P.J., Stewart P.R. Mitochondrial and peroxisomal contributions to the energy metabolism of Saccharomyces cerevisiae in continuous culture. J Gen Microbiol 1973; 79:205–217
    [Google Scholar]
  31. Ryan E.D., Kohlhaw G.B. Subcellular localization of isoleucine-valine biosynthetic enzymes in yeast. J Bacteriol 1974; 120:631–637
    [Google Scholar]
  32. Schatz G., Mason T. The biosynthesis of mitochondrial proteins. Annu Rev Biochem 1974; 43:51–87
    [Google Scholar]
  33. Shimizu I., Nagai J., Hatanaka H., Katsuki H. Mevalonate synthesis in the mitochondria of yeast. Biochim Biophys Acta 1973; 296:310–320
    [Google Scholar]
  34. Stuart R.A., Gruhler A., Van der Klei I., Guiard B., Koll H., Neupert W. The requirement of matrix ATP for the import of precursor proteins into the mitochondrial matrix and inter-membrane space. Eur J Biochem 1994; 220:9–18
    [Google Scholar]
  35. Šubík J., Kolarov J., Kováč L.K. Obligatory requirement of intramitochondrial ATP for normal functioning of the eukaryotic cell. Biochem Biophys Res Commun 1972; 49:192–198
    [Google Scholar]
  36. van Urk H., Bruinenberg P.M., Veenhuis M., Scheffers W.A., van Dijken J.P. Respiratory capacities of mitochondria of Saccharomyces cerevisiae CBS 8066 and Candida utilis CBS 621 grown under glucose limitation. Antonie Leeuwenhoek 1989; 56:211–220
    [Google Scholar]
  37. Veenhuis M., Harder W. Metabolic significance and biogenesis of microbodies in yeasts. In Peroxisomes in Biology and Medicine 1987 Edited by Fahimi H.D., Sies H. Berlin: Springer Verlag; pp 436–458
    [Google Scholar]
  38. Verduyn C., Postma E., Scheffers W.A., van Dijken J.P. Energetics of Saccharomyces cerevisiae in anaerobic glucose-limited chemostat cultures. J Gen Microbiol 1990; 136:405–412
    [Google Scholar]
  39. Verduyn C., Postma E., Scheffers W.A., van Dijken J.P. Effect of benzoic acid on metabolic fluxes in yeascs: a continuous culture study on the regulation of respiration and alcoholic fermentation. Yeast 1992; 8:501–517
    [Google Scholar]
  40. Wales D.S., Cartledge T.G., Lloyd D. Effects of glucose repression and anaerobiosis on the activities and subcellular distribution of tricarboxylic acid cycle and associated enzymes in Saccharomyces carlsbergensis. J Gen Microbiol 1980; 116:93–98
    [Google Scholar]
  41. Wallace P.G., Linnane A.W. Oxygen-induced synthesis of yeast mitochondria. Nature 1964; 201:1191–1194
    [Google Scholar]
  42. Walsh K., Koshland D.E. Jr Characterization of rate- controlling steps in vivo by the use of an adjustable expression vector. Proc Natl Acad Sci USA 1985; 82:3577–3581
    [Google Scholar]
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