Skip to main content

Cytoduction: A tool for mitochondrial genetic studies in yeast

Utilization of the nuclear-fusion mutation kar 1-1 for transfer of drug r and mit genomes in Saccharomyces cerevisiae

Summary

A study has been made of the general applications of the nuclear-fusion mutation, kar1, to mitochondrial genetic research. Procedures were developed which are suitable for constructing new strains by transfer (cytoduction) of mitochondrial genomes containing drug r, mit -, syn - or rho - mutations.

Several examples of crosses of the type KAR rho + drug r xkarl rho o drug s were carried out and the resulting zygotes and their first buds separated by micromanipulation. Clones derived from these were in most cases homogeneous for any of the following nuclear types: heterokaryon (a+α), diploid (a/α), haploid a nucleus or haploid α nucleus. The term cytoductant is given to the haploid rho + segregant having the same type of nucleus as the rho o strain employed in the cross.

As examples of cytoduction of mit - mutations, two different pho mitochondrial genomes were studied. Transfer of pho to rho o acceptors was achieved with little difficulty.

Haploid segregants containing recombinant mitochondrial genomes were obtained by crossing a KAR rho + cap r oli j par r strain with a karl rho + cap s oli s par s strain. Studies of intraclonal distribution of alleles indicate that cytoplasmic mixing is restricted when the dikaryotic state is maintained. Haploid cytoductant clones are usually comprised of cells retaining the mitochondrial genome of only one parent, and commonly both first-bud and residual-zygote clones are homogeneous for the same parental genome (apparent uniparental inheritance). Less frequently mixed clones are found where the number of classes of mitochondrial recombinant type is limited compared to the progeny of zygotes in which nuclear fusion takes place.

These observations are discussed in terms of number and form of mitochondria forming the chondriome in the yeast cell, together with its association with the nucleus.

This is a preview of subscription content, access via your institution.

References

  1. Beck, J.C., Mattoon, J.R., Hawthorne, D.C., Sherman, F.: Genetic modification of energy-conserving system in yeast mitochondria. Proc. Natl. Acad. Sci. (Wash.) 60, 186–193 (1968)

    Google Scholar 

  2. Birky, C.W., Jr., Demko, C.A., Perlman, P.S., Strauberg, R.: Uniparental inheritance of mitochondrial genes in yeast: Dependence on input bias of mitochondrial DNA and preliminary investigations of the mechanism. Genetics 89, 615–651 (1978)

    Google Scholar 

  3. Coen, D., Deutsch, J., Netter, P., Petrochilo, E., Slonimski, P.P.: Mitochondrial genetics. I. Methodology and phenomenology. Symp. Soc. exp. Biol. 24, 449–496 (1970)

    Google Scholar 

  4. Colson, A.-M., Goffeau, A., Briquet, M., Weigel, P., Mattoon, J.R.: Nucleo-cytoplasmic interaction between oligomycin-resistant mutations in Saccharomyces cerevisiae. Molec. gen. Genet. 135, 309–326 (1974)

    Google Scholar 

  5. Conde, J., Fink, G.R.: A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc. Natl. Acad. Sci. (Wash.) 73, 3651–3655 (1976)

    Google Scholar 

  6. Douglas, H.C., Pelroy, G.: A gene controlling inducibility of the galactose pathway enzymes in Saccharomyces. Biochim. Biophys. Acta 68, 155–156 (1963)

    Google Scholar 

  7. Dujon, B., Slonimski, P.P., Weill, L.: Mitochondrial genetics IX: A model for recombinationand segregation of mitochondrial genomes in Saccharomyces cerevisiae. Genetics 78, 415–437 (1974)

    Google Scholar 

  8. Foury, F., Tzagoloff, A.: Localization on mitochondrial DNA of mutations leading to a loss of rutamycin-sensitive adenosine triphosphatase. Eur. J. Biochem. 68, 113–119 (1976)

    Google Scholar 

  9. Grimes, G.W., Mahler, H.R., Perlman, P.S.: Nuclear gene dosage effects on mitochondorial mass and DNA. J. Cell. Biol. 61, 565–574 (1974)

    Google Scholar 

  10. Gunge, N., Sakaguchi, K.: Fusion of mitochondria with protoplasts in yeast. In: Abs. 9th Int. Conf. Yeast Genetics and Molecular Biology, p. 34, 1978

  11. Hartwell, L.H.: Synchronization of haploid yeast cell cycle, a prelude to conjugation. Exp. Cell. Res. 76, 111–117 (1973)

    Google Scholar 

  12. Kotylak, Z., Slonimski, P.P.: Joint control of cytochromes a and b by a unique mitochondrial DNA region comprising four genetic loci. In: The Genetic Function of Mitochondrial DNA (C. Saccone and A.M. Kroon, eds.), pp. 143–154. Amsterdam: North-Holland 1976

    Google Scholar 

  13. Lancashire, W.E.: A genetic approach to oxidative phosphorylation. Ph.D. Thesis, University of Warwick, England (1974)

  14. Lancashire, W.E.: Mitochondrial mutations conferring hear or cold sensitivity in Saccharomyces cerevisiae. In: Genetics and Biogenesis of Chloroplast and Mitochondria (Th. Bücher, W. Neupert, W. Sebald and S. Werner, eds.), pp. 481–490, Amsterdam: North-Holland 1976

    Google Scholar 

  15. Lancashire, W.F., Griffiths, D.E.: Studies on energy-linked reactions: Isolation, characterisation and genetic analysis of trialkyltin resistant mutants of Saccharomyces cerevisiae. Eur. J. Biochem. 51, 377–392 (1975)

    Google Scholar 

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

    Google Scholar 

  17. Mattoon, J.R., Malamud, D.R., Brunner, A., Braz, G., Carvajal, E., Lancashire, W.E., Panek, A.D.: Regulation of heme formation and cytochrome biosynthesis in normal and mutant yeast. In: Biochemistry and genetics of yeast, pure and applied aspects (M. Bacila, B. Horecker and A.O.M. Stoppani, eds.). pp. 317–337 New York: Academic Press 1978

    Google Scholar 

  18. Ortega Ruiz, J.M.: Transformation of respiratory deficient strains of Saccharomyces cerevisiae with mitochondria obtained from respiratory normal strains. Microbios. Letts. 4, 133–137 (1977)

    Google Scholar 

  19. Sanders, H.K., Mied, P.A., Briquet, M., Hernandez-Rodriguez, J., Gottal, R.F., Mattoon, J.R.: Regulation of mitochondrial biogenesis: Yeast mutants deficient in synthesis of δ-aminolevulinic acid. J. Mol. Biol. 80, 17–39 (1973)

    Google Scholar 

  20. Sanders, J.P.M., Heyting, C., Verbeet, M.Ph., Meijlink, F.C.P.W., Borst, P.: The organization of genes in yeast mitochondrial DNA. III. Comparison of the physical maps of the mitochondrial DNAs from three wild-type Saccharomyces strains. Molec. gen. Genet. 157, 239–261 (1977)

    Google Scholar 

  21. Schweyen, R.J., Weiss-Brummer, B., Backhaus, B., Kaudewitz, F.: The genetic map of the mitochondrial genome in yeast. Map positions of drug r and mit - markers as revealed from population analyses of rho - clones in Saccharomyces cerevisiae. Molec. gen. Genet. 159, 151–160 (1978)

    Google Scholar 

  22. Stevens, B.J.: Variation in number and volume of the mitochondria in yeast according to growth conditions. A study based on serial sectioning and computer graphics reconstruction. Biologie Cellulaire 28, 37–56 (1977)

    Google Scholar 

  23. Strausberg, R.L., Perlman, P.S.: The effect of zygotic bud position on the transmission of mitochondrial genes in Saccharomyces cerevisiae. Molec. gen. Genet. 163, 131–144 (1978)

    Google Scholar 

  24. Thomas, D.Y., Wilkie, D.: Recombination of mitochondrial drugresistance factors in Saccharomyces cerevisiae. Biochem. Biophys. Res. Commun. 30, 368–372 (1968)

    Google Scholar 

  25. Trembath, M.K., Macino, G., Tzagoloff, A.: The mapping of mutations in tRNA and cytochrome oxidase genes located in the cap-par segment of the mitochondrial genome of S. cerevisiae. Molec. gen. Genet. 158, 35–45 (1977)

    Google Scholar 

  26. Tzagoloff, A., Akai, A., Needleman, R.B., Zulch, G.: Assembly of the mitochondrial membrane system: Cytoplasmic mutants of Saccharomyces cerevisiae with lesions in enzymes of the respiratory chain and in mitochondrial ATPase. J. Biol. Chem. 250, 8236–8242 (1975)

    Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to William E. Lancashire.

Additional information

Communicated by F. Kaudewitz

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Lancashire, W.E., Mattoon, J.R. Cytoduction: A tool for mitochondrial genetic studies in yeast. Molec. Gen. Genet. 170, 333–344 (1979). https://doi.org/10.1007/BF00267067

Download citation

Keywords

  • Yeast Cell
  • Genetic Study
  • Mitochondrial Genome
  • Genetic Research
  • General Application