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Mitochondrial DNA replication in petite mutants of yeast: Resistance to inhibition by ethidium bromide, berenil and euflavine


Mitochondrial DNA (mtDNA) replication in petite mutants ofSaccharomyces cerevisiae is generally less sensitive to inhibition by ethidium bromide than in grande (respiratory competent) cells. In every petite that we have examined, which retain a range of different grande mtDNA sequences, this general phenomenon has been demonstrated by measurements of the loss of mtDNA from cultures grown in the presence of the drug. The resistance is also demonstrable by direct analysis of drug inhibition of mtDNA replication in isolated mitochondria. Furthermore, the resistance to ethidium bromide is accompanied, in every case tested, by cross-resistance to berenil and euflavine, although variations in the levels of resistance are observed.

In one petite the level of in vivo resistance to the three drugs was very similar (4-fold over the grande parent) whilst another petite was mildly resistant to ethidium bromide and berenil (each 1.6-fold over the parent) and strongly resistant (nearly 8-fold) to inhibition of mtDNA replication by euflavine. The level of resistance to ethidium bromide in several other petite clones tested was found to vary markedly. Using genetic techniques it is possible to identify those petites which display an enhanced resistance to ethidium bromide inhibition of mtDNA replication.

It is considered that the general resistance of petites arises because a product of mitochondrial protein synthesis is normally involved in facilitating the inhibitory action of these drugs on mtDNA synthesis in grande cells. The various levels of resistance in petites may be modulated by the particular mtDNA sequences retained in each petite.

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  1. Bastos, R.N.: Use of diazidoethidium bromide as a specific probe for mitochondrial functions. J. biol. Chem.250, 7739–7746 (1975)

  2. Bastos, R.N., Mahler, H.R.: Molecular mechanisms of mitochondrial genetic activity. Effects of ethidium bromide on the deoxyribonucleic acid and energetics of isolated mitochondria. J. biol. Chem.249, 6617–6627 (1974)

  3. Borst, P.: Mitochondrial nucleic acids. Ann. Rev. Biochem.41, 333–376 (1972)

  4. Goldring, E.S., Grossman, L.I., Krupnick, D., Cryer, D.R., Marmur, J.: The petite mutation in yeast. I. The loss of mitochondrial DNA during petite induction with ethidium bromide. J. molec. Biol.52, 323–335 (1970)

  5. Hall, R.M., Mattick, J.S., Marzuki, S., Linnane, A.W.: Evidence for a functional association of DNA synthesis with the membrane in mitochondria ofSaccharomyces cerevisiae. Molec. Biol. Reports2, 101–106 (1975)

  6. Hall, R.M., Nagley, P., Linnane, A.W.: Biogenesis of mitochondria. 42. Genetic analysis of the control of mitochondrial DNA levels inSaccharomyces cerevisiae. Molec. gen. Genet.145, 169–175 (1976)

  7. Hollenberg, C.P., Borst, P., Flavell, R.A., Van Kreijl, C.F., Van Bruggen, E.F.J., Arnberg, A.C.: The unusual properties of mtDNA from a “low density” petite mutant of yeast. Biochim. biophys. Acta (Amst.)277, 44–58 (1972a)

  8. Hollenberg, C.P., Borst, P., Van Bruggen, E.F.J.: Mitochondrial DNA from cytoplasmic petite mutants of yeast. Biochim. biophys. Acta (Amst.)277, 35–43 (1972b)

  9. Linnane, A.W., Haslam, J.M., Lukins, H.B., Nagley, P.: The biogenesis of mitochondria in microorganisms. Ann. Rev. Microbiol.26, 163–198 (1972)

  10. Linnane, A. W., Lukins, H.B., Molloy, P.L., Nagley, P., Rytka, J., Sriprakash, K.S., Trembath, M.K.: Biogenesis of mitochondria. 45. Molecular mapping of the mitochondrial genome of yeast. Proc. nat. Acad. Sci. (Wash.)73, 2082–2085 (1976)

  11. Linnane, A.W., Saunders, G.W., Gingold, E.B., Lukins, H.B.: The biogenesis of mitochondria. V. Cytoplasmic inheritance of erythromycin resistance inSaccharomyces cerevisiae. Proc. nat. Acad. Sci. (Wash.)59, 903–910 (1968)

  12. Mahler, H.R.: Biogenetic autonomy of mitochondria. CRC Crit. Rev. Biochem.1, 381–460 (1973a)

  13. Mahler, H.R.: Genetic autonomy of mitochondrial DNA. In: Molecular cytogenetics (Hamkalo, B. and Papaconstantinou, J., eds.), pp. 181–208. New York: Plenum Press 1973b

  14. Mahler, H.R.: Mitochondrial assembly: attempts at resolution of complex functions. In: Mitochondria: bioenergetics, biogenesis and membrane structure (Packer, L. and Gomez-Puyou, A., eds.), pp. 213–240. New York: Academic Press 1976

  15. Mattick, J.S., Hall, R.M.: Replicative DNA synthesis in isolated mitochondria ofSaccharomyces cerevisiae. J. Bact. (in press, 1977)

  16. Mattick, J. S., Nagley, P.: Comparative studies on the effects of acridines and other petite inducing drugs on the mitochondrial genome ofSaccharomyces cerevisiae. Molec. gen. Genet.152, 267–276 (1977)

  17. Mol, J.N.M., Borst, P., Grosveld, F.G., Spencer, J.H.: The size of the repeating unit of the repetitive mitochondrial DNA from a “low density” petite mutant of yeast. Biochim. biophys. Acta (Amst.)474, 115–128 (1974)

  18. Molloy, P.L., Linnane, A.W., Lukins, H.B.: Biogenesis of mitochondria. 37. Analysis of deletion of antibiotic resistance markers in petite mutants ofSaccharomyces cerevisiae. J. Bact.122, 7–18 (1975)

  19. Nagley, P., Gingold, E.B., Linnane, A.W.: The purification of mitochondrial genes using petite mutants of yeast. In: The biogenesis of mitochondria: transcriptional, translational and genetic aspects (Kroon, A.M. and Saccone, C., eds.), pp. 157–168. New York: Academic Press 1974a

  20. Nagley, P., Gingold, E. B., Lukins, H. B., Linnane, A.W.: Biogenesis of mitochondria. XXV. Studies on the mitochondrial genomes of petite mutants of yeast using ethidium bromide as a probe. J. molec. Biol.78, 335–350 (1973)

  21. Nagley, P., Linnane, A.W.: Biogenesis of mitochondria. XXI. Studies on the nature of the mitochondrial genome in yeast: the degenerative effects of ethidium bromide on mitochondrial genetic information in a respiratory competent strain. J. molec. Biol.66, 181–193 (1972)

  22. Nagley, P., Mattick, J.S., Hall, R.M., Linnane, A.W.: Biogenesis of mitochondria. 43. A comparative study of petite induction and inhibition of mitochondrial DNA replication by ethidium bromide and berenil. Molec. gen. Genet.141, 291–304 (1975)

  23. Nagley, P., Molloy, P.L., Lukins, H.B., Linnane, A.W.: Studies on mitochondrial gene purification using petite mutants of yeast. Characterization of mutants enriched in ribosomal RNA cistrons. Biochem. biophys. Res. Commun.57, 232–239 (1974b)

  24. Nagley, P., Sriprakash, K.S., Linnane, A.W.: Structure synthesis and genetics of yeast mitochondrial DNA. In: Advances in microbial physiology (Rose, A. H. and Morris, J.G., eds.). New York: Academic Press (in press, 1977)

  25. Perlman, P.S., Mahler, H.R.: Molecular consequences of ethidium bromide mutagenesis. Nature (Lond.) New Biol.231, 12–16 (1971)

  26. Schatz, G., Mason, T.L.: The biosynthesis of mitochondrial proteins. Ann. Rev. Biochem.43, 51–87 (1974)

  27. Sriprakash, K.S., Molloy, P.L., Nagley, P., Lukins, H.B., Linnane, A.W.: Biogenesis of mitochondria. XLI. Physical mapping of mitochondrial genetic markers in yeast. J. molec. Biol.104, 485–503 (1976)

  28. Trembath, M.K., Molloy, P.L., Cutting, G.L., Skriprakash, K.S., Linnane, A.W., Lukins, H.B.: Biogenesis of mitochondria. 44. Comparative studies and mapping of mitochondrial oligomycin resistance mutations in yeast based on gene recombination and petite deletion analysis. Molec. gen. Genet.145, 43–52 (1976)

  29. Waring, M.: Ethidium and propidium. In: Antibiotics III. Mechanism of action of antimicrobial and antitumor agents (Corcoran, J.W. and Hahn, F.E., eds.), pp. 141–165. Berlin-Heidelberg-New York: Springer 1975

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Nagley, P., Mattick, J.S. Mitochondrial DNA replication in petite mutants of yeast: Resistance to inhibition by ethidium bromide, berenil and euflavine. Molec. Gen. Genet. 152, 277–283 (1977). https://doi.org/10.1007/BF00693081

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  • Protein Synthesis
  • Inhibitory Action
  • General Resistance
  • Ethidium Bromide
  • Mitochondrial Protein