Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1970 Jul 1;46(1):88–96. doi: 10.1083/jcb.46.1.88

BIOGENESIS OF MITOCHONDRIA

XIII. The Isolation of Mitochondrial Structures from Anaerobically Grown Saccharomyces cerevisiae

K Watson 1, J M Haslam 1, Anthony W Linnane 1
PMCID: PMC2108061  PMID: 4248186

Abstract

Morphologically intact structures have been isolated from anaerobically grown yeast cells which have many of the properties of yeast mitochondria. The structures are about 0.5 µ in diameter and contain malate dehydrogenase, succinate dehydrogenase, oligomycin-sensitive ATPase, and DNA of buoyant density 1.683 g/cc, characteristic of yeast mitochondria. The morphology of the structures is critically dependent on their lipid composition. When isolated from cells grown anaerobically in the presence of supplements of unsaturated fatty acid and ergosterol, their unsaturated fatty acid content is similar to that of mitochondria from aerobically grown cells. These lipid-complete structures consist pre-dominantly of double-membrane vesicles enclosing a dense matrix which contains a folded inner membrane system bordering electron-transparent regions which are somewhat different from the cristae of functional mitochondria. In contrast, the structures from cells grown without lipid supplements are much simpler in morphology; they have a dense granular matrix surrounded by a double membrane but have no obvious folded inner membrane system within the matrix. The lipid-depleted structures are very fragile and are only isolated in intact form from protoplasts that have been prefixed with glutaraldehyde

Full Text

The Full Text of this article is available as a PDF (896.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ARRIGONI O., SINGER T. P. Limitations of the phenazine methosulphate assay for succinic and related dehydrogenases. Nature. 1962 Mar 31;193:1256–1258. doi: 10.1038/1931256a0. [DOI] [PubMed] [Google Scholar]
  2. BLOOMFIELD D. K., BLOCH K. The formation of delta 9-unsaturated fatty acids. J Biol Chem. 1960 Feb;235:337–345. [PubMed] [Google Scholar]
  3. Criddle R. S., Schatz G. Promitochondria of anaerobically grown yeast. I. Isolation and biochemical properties. Biochemistry. 1969 Jan;8(1):322–334. doi: 10.1021/bi00829a045. [DOI] [PubMed] [Google Scholar]
  4. DUELL E. A., INOUE S., UTTER M. F. ISOLATION AND PROPERTIES OF INTACT MITOCHONDRIA FROM SPHEROPLASTS OF YEAST. J Bacteriol. 1964 Dec;88:1762–1773. doi: 10.1128/jb.88.6.1762-1773.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Damsky C. H., Nelson W. M., Claude A. Mitochondria in anaerobically-grown, lipid-limited brewer's yeast. J Cell Biol. 1969 Oct;43(1):174–179. doi: 10.1083/jcb.43.1.174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FUKUHARA H. RNA SYNTHESIS OF YEAST IN THE PRESENCE OF CYCLOHEXIMIDE. Biochem Biophys Res Commun. 1965 Jan 18;18:297–301. doi: 10.1016/0006-291x(65)90757-6. [DOI] [PubMed] [Google Scholar]
  7. GREEN D. E., FLEISCHER S. THE ROLE OF LIPIDS IN MITOCHONDRIAL ELECTRON TRANSFER AND OXIDATIVE PHOSPHORYLATION. Biochim Biophys Acta. 1963 Oct 22;70:554–582. doi: 10.1016/0006-3002(63)90793-5. [DOI] [PubMed] [Google Scholar]
  8. Green D. E., Asai J., Harris R. A., Penniston J. T. Conformational basis of energy transformations in membrane systems. 3. Configurational changes in the mitochondrial inner membrane induced by changes in functional states. Arch Biochem Biophys. 1968 May;125(2):684–705. doi: 10.1016/0003-9861(68)90626-7. [DOI] [PubMed] [Google Scholar]
  9. Hackenbrock C. R. Chemical and physical fixation of isolated mitochondria in low-energy and high-energy states. Proc Natl Acad Sci U S A. 1968 Oct;61(2):598–605. doi: 10.1073/pnas.61.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hackenbrock C. R. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. I. Reversible ultrastructural changes with change in metabolic steady state in isolated liver mitochondria. J Cell Biol. 1966 Aug;30(2):269–297. doi: 10.1083/jcb.30.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hackenbrock C. R. Ultrastructural bases for metabolically linked mechanical activity in mitochondria. II. Electron transport-linked ultrastructural transformations in mitochondria. J Cell Biol. 1968 May;37(2):345–369. doi: 10.1083/jcb.37.2.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris R. A., Penniston J. T., Asai J., Green D. E. The conformational basis of energy conservation in membrane systems. II. Correlation between conformational change and functional states. Proc Natl Acad Sci U S A. 1968 Mar;59(3):830–837. doi: 10.1073/pnas.59.3.830. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jayaraman J., Cotman C., Mahler H. R., Sharp C. W. Biochemical correlates of respiratory deficiency. VII. Glucose repression. Arch Biochem Biophys. 1966 Sep 26;116(1):224–251. doi: 10.1016/0003-9861(66)90029-4. [DOI] [PubMed] [Google Scholar]
  14. Jollow D., Kellerman G. M., Linnane A. W. The biogenesis of mitochondria. 3. The lipid composition of aerobically and anaerobically grown Saccharomyces cerevisiae as related to the membrane systems of the cells. J Cell Biol. 1968 May;37(2):221–230. doi: 10.1083/jcb.37.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Korn E. D. Cell membranes: structure and synthesis. Annu Rev Biochem. 1969;38:263–288. doi: 10.1146/annurev.bi.38.070169.001403. [DOI] [PubMed] [Google Scholar]
  16. LINDENMAYER A., ESTABROOK R. W. Low-temperature spectral studies on the biosynthesis of cytochromes in Baker's yeast. Arch Biochem Biophys. 1958 Nov;78(1):66–82. doi: 10.1016/0003-9861(58)90315-1. [DOI] [PubMed] [Google Scholar]
  17. Lamb A. J., Clark-Walker G. D., Linnane A. W. The biogenesis of mitochondria. 4. The differentiation of mitochondrial and cytoplasmic protein synthesizing systems in vitro by antibiotics. Biochim Biophys Acta. 1968 Jul 23;161(2):415–427. [PubMed] [Google Scholar]
  18. Lukins H. B., Tham S. H., Wallace P. G., Linnane A. W. Correlation of membrane bound succinate dehydrogenase with the occurrence of mitochondrial profiles in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1966 May 25;23(4):363–367. doi: 10.1016/0006-291x(66)90734-0. [DOI] [PubMed] [Google Scholar]
  19. MORPURGO G., SERLUPI-CRESCENZI G., TECCE G., VALENTE F., VENETTACCI D. INFLUENCE OF ERGOSTEROL ON THE PHYSIOLOGY AND THE ULTRA-STRUCTURE OF SACCHAROMYCES CEREVISIAE. Nature. 1964 Feb 29;201:897–899. doi: 10.1038/201897a0. [DOI] [PubMed] [Google Scholar]
  20. PULLMAN M. E., MONROY G. C. A NATURALLY OCCURRING INHIBITOR OF MITOCHONDRIAL ADENOSINE TRIPHOSPHATASE. J Biol Chem. 1963 Nov;238:3762–3769. [PubMed] [Google Scholar]
  21. Paltauf F., Schatz G. Promitochondria of anaerobicallly grown yeast. II. Lipid composition. Biochemistry. 1969 Jan;8(1):335–339. doi: 10.1021/bi00829a046. [DOI] [PubMed] [Google Scholar]
  22. Penniston J. T., Harris R. A., Asai J., Green D. E. The conformational basis of energy transformations in membrane systems. I. Conformational changes in mitochondria. Proc Natl Acad Sci U S A. 1968 Feb;59(2):624–631. doi: 10.1073/pnas.59.2.624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Plattner H., Schatz G. Promitochondria of anaerobically grown yeast. 3. Morphology. Biochemistry. 1969 Jan;8(1):339–343. doi: 10.1021/bi00829a047. [DOI] [PubMed] [Google Scholar]
  24. Proudlock J. W., Haslam J. M., Linnane A. W. Specific effect of unsaturated fatty acid depletion on mitochondrial oxidative phosphorylation in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1969 Nov 20;37(5):847–852. doi: 10.1016/0006-291x(69)90969-3. [DOI] [PubMed] [Google Scholar]
  25. SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. SCHATZ G. SUBCELLULAR PARTICLES CARRYING MITOCHONDRIAL ENZYMES IN ANAEROBICALLY-GROWN CELLS OF SACCHAROMYCES CEREVISIAE. Biochim Biophys Acta. 1965 Feb 22;96:342–345. [PubMed] [Google Scholar]
  27. SCHILDKRAUT C. L., MARMUR J., DOTY P. Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. J Mol Biol. 1962 Jun;4:430–443. doi: 10.1016/s0022-2836(62)80100-4. [DOI] [PubMed] [Google Scholar]
  28. Tustanoff E. R., Bartley W. Development of respiration in yeast grown anaerobically on different carbon sources. Biochem J. 1964 Jun;91(3):595–600. doi: 10.1042/bj0910595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Vary M. J., Edwards C. L., Stewart P. R. The biogenesis of mitochondria. IX. Formation of the soluble mitochondrial enzymes malate dehydrogenase and fumarase in Saccharomyces cerevisiae. Arch Biochem Biophys. 1969 Mar;130(1):235–243. doi: 10.1016/0003-9861(69)90029-0. [DOI] [PubMed] [Google Scholar]
  30. WALLACE P. G., LINNANE A. W. OXYGEN-INDUCED SYNTHESIS OF YEAST MITOCHONDRIA. Nature. 1964 Mar 21;201:1191–1194. doi: 10.1038/2011191a0. [DOI] [PubMed] [Google Scholar]
  31. Wake R. G. A study of the possible extent of synthesis of repair DNA during germination of Bacillus subtilis spores. J Mol Biol. 1967 Apr 28;25(2):217–234. doi: 10.1016/0022-2836(67)90139-8. [DOI] [PubMed] [Google Scholar]
  32. Wallace P. G., Huang M., Linnane A. W. The biogenesis of mitochondria. II. The influence of medium composition on the cytology of anaerobically grown Saccharomyces cerevisiae. J Cell Biol. 1968 May;37(2):207–220. doi: 10.1083/jcb.37.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. YOTSUYANAGI Y. [Study of yeast mitochondria. II. Mitochondria of respiration-deficient mutants]. J Ultrastruct Res. 1962 Aug;7:141–158. doi: 10.1016/s0022-5320(62)80032-x. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES