1887

Abstract

Summary: Two methods were used to isolate mitochondria from In the first method, cells were weakened by exposure to hypotonic conditions and then disrupted by blending; mitochondria were subsequently isolated using disodium 3,5-diacetoamido-2,4,6-triiodobenzoate gradients. In the second, cells were treated with digitonin before disruption; mitochondria were purified by differential centrifugation. Both preparations were examined with the electron microscope and were also shown to possess several characteristic biochemical properties of mitochondria. Kinetoplast DNA was present in the mitochondria, uncontaminated by nuclear DNA.

Analysis by polyacrylamide gel electrophoresis showed two RNA components of molecular weights 0·47 × 10 and 0·22 × 10, in addition to cytoplasmic RNA contamination. Four mitochondrial components with sedimentation coefficients of 14·6S, 11·4S, 10·1S and 6·9S were identified on sucrose density gradients. Ethidium bromide abolished the incorporation of [5-H] uridine into the presumed mitochondrial RNA.

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1977-04-01
2022-05-28
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References

  1. Attardi G., Aloni Y., Attardi B., Ojala D., Pica-Mattoccia L., Robbertson D. L., Storrie B. 1970; Transcription of mitochondrial DNA in HeLa cells. Cold Spring Harbor Symposia on Quantitative Biology 35:599–619
    [Google Scholar]
  2. Borst P. 1972; Mitochondrial nucleic acids. Annual Review of Biochemistry 41:333–376
    [Google Scholar]
  3. Braly P., Simpson L., Kretzer F. 1974; Isolation of kinetoplast-mitochondrial complexes from Leishmania tarentolae. Journal of Protozoology 21:782–790
    [Google Scholar]
  4. Bresslau E., Scremin L. 1924; Die Kerne der Trypanosomen und ihr Verhalten zur Nuclealreaktion. Archiv für Protistenkunde 48:509–515
    [Google Scholar]
  5. Dubuy H. G., Mattern C. F., Riley F. L. 1965; Isolation and characterization of DNA from kinetoplasts of Leishmania enriettii. Science 147:754–756
    [Google Scholar]
  6. Hill G. C., White D. W. 1968; Respiratory pigments of Crithidia fasciculata. Journal of Bacteriology 95:2151–2157
    [Google Scholar]
  7. Kallinikova V. D. 1969; Functional value of trypanosomatid kinetoplast in the light of cytochemical investigations. In Progress in Protozoology IIIrd International Congress on Protozoology pp. 31–32
    [Google Scholar]
  8. Kidder G. W., Dutta B. N. 1958; The growth and nutrition of Crithidia fasciculata. Journal of General Microbiology 18:621–635
    [Google Scholar]
  9. King T. W. 1967; Preparation of succinate dehydrogenase and reconstitution of succinate oxidase. Methods in Enzymology 10:322–331
    [Google Scholar]
  10. Kirby K. S. 1965; Isolation and characterization of ribosomal ribonucleic acids. Biochemical Journal 96:266–269
    [Google Scholar]
  11. Kusel J. P., Storey B. T. 1972; Evidence for the presence of two phosphorylation sites in mitochondria isolated from the trypanosomatid hemofiagellate, Crithidia fasciculata. Biochemical and Biophysical Research Communications 46:501–507
    [Google Scholar]
  12. Langdon R. 1966; Glucose-6-phosphate dehydrogenase from erythrocytes. Methods in Enzymology 9:126–131
    [Google Scholar]
  13. Loening U. 1967; The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochemical Journal 102:251–257
    [Google Scholar]
  14. Loening U. 1969; The determination of the molecular weight of ribonucleic acid by polyacrylamide gel electrophoresis. The effects of changes in conformation. Biochemical Journal 113:131–138
    [Google Scholar]
  15. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  16. Mancilla R., Náquira C. 1964; Comparative metabolism of 14C-glucose in two strains of Trypanosoma cruzi. Journal of Protozoology 11:509–513
    [Google Scholar]
  17. Ozeki Y., Sooksri V., Ono T., Inoki S. 1971; Studies on the ultrastructure of kinetoplasts of Trypanosoma cruzi and Trypanosoma gambiense by autoradiography and enzymatic digestion. Bikeris Journal 14:97–118
    [Google Scholar]
  18. Pace N. R. 1973; Structure and synthesis of ribosomal ribonucleic acid of prokaryotes. Bacteriological Reviews 37:562–603
    [Google Scholar]
  19. Parish J. H., Kirby K. S. 1966; Reagents which reduce interactions between ribosomal RNA and rapidly labelled RNA. Biochimica et biophysica acta 129:554–562
    [Google Scholar]
  20. Paulin J. J. 1975; The chondriome of selected trypanosomatids. A three-dimensional study based on serial thick sections and high voltage electron microscopy. Journal of Cell Biology 66:404–413
    [Google Scholar]
  21. Render H., Wolstenholme D. 1970; Kinetoplast deoxyribonucleic acid of the hemoflagellate Trypanosoma lewisi. Journal of Cell Biology 47:689–702
    [Google Scholar]
  22. Renger H., Wolstenholme D. 1972; The form and structure of kinetoplast DNA of Crithidia. Journal of Cell Biology 54:346–364
    [Google Scholar]
  23. Reijnders L., Sloof P., Sival J., Borst P. 1973; Gel electrophoresis of RNA under denaturing conditions. Biochimica et biophysica acta 324:320–333
    [Google Scholar]
  24. Schildkraut C. L., Marmur J., Doty P. 1962; Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl. Journal of Molecular Biology 4:430–443
    [Google Scholar]
  25. Simpson L. 1968; Behaviour of kinetoplast of Leishmania tarentolae upon cell rupture. Journal of Protozoology 15:132–136
    [Google Scholar]
  26. Simpson L. 1972; The kinetoplast of the hemoflagellates. International Review of Cytology 32:139–207
    [Google Scholar]
  27. Simpson L. 1973; Structure and function of kinetoplast DNA. Journal of Protozoology 20:2–8
    [Google Scholar]
  28. Simpson L., Lasky L. 1975; Kinetoplast messenger RNA’s. Journal of Cell Biology 67:402a
    [Google Scholar]
  29. Spencer R., Cross G. A. M. 1976; Lability of RNA from the large cytoplasmic ribosomal subunit of the protozoon Crithidia oncopelti. Journal of General Microbiology 93:82–88
    [Google Scholar]
  30. Steinert M. 1960; Mitochondria associated with the kinetonucleus of Trypanosoma mega. Journal of Biophysical and Biochemical Cytology 8:542–546
    [Google Scholar]
  31. Steinert M., Van Assel S. 1975; L’ADN mitochondrial de Crithidia. Journal of Protozoology 22:85a
    [Google Scholar]
  32. Steinert G., Firket H., Steinert M. 1958; Synthèse d’acide désoxyribonucldique dans le corps parabasal de Trypanosoma mega. Experimental Cell Research 15:632–635
    [Google Scholar]
  33. Steinert M., Van Assel S., Steinert G. 1969; Étude, par autoradiographie, des effets du bromure d’éthidium sur la synthèse des acides nucléiques de Crithidia luciliae. Experimental Cell Research 56:69–74
    [Google Scholar]
  34. Szybalski W. 1968; Equilibrium sedimentation of viruses, nucleic acids and other macromolecules in density gradients. Fractions 1:1–15 Beckman Instruments Inc.
    [Google Scholar]
  35. Toner J. J., Weber M. M. 1972; Respiratory control in mitochondria from Crithidia fasciculata. Biochemical and Biophysical Research Communications 46:652–660
    [Google Scholar]
  36. Vesco C., Penman S. 1969; The cytoplasmic RNA of HeLa cells: new discrete species associated with mitochondria. Proceedings of the National Academy of Sciences of the United States of America 62:218–225
    [Google Scholar]
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