Summary
The isolation and characterization of five new mutants affecting mitochondrial protein synthesis in S. cerevisiae is reported. Each mutation confers in vivo resistance to the macrolide antibiotic spiramycin which acts by inhibiting mitochondrial protein synthesis in sensitive yeast. The mutants are distinguishable on the basis of their in vivo cross resistance to other antibiotics, their biochemical properties and genetic behaviour. Genetic analysis indicates the mode of inheritance to be nuclear for one mutation and cytoplasmic for the other four. Recombination analysis performed on crosses between different cytoplasmic determinants, together with data from monofactorial crosses of each determinant with sensitive strains, demonstrates at least two and possibly three cytoplasmic genetic loci conferring spiramycin resistance.
The protein synthesizing activities of mitochondria isolated from the mutant strains range in response to spiramycin and other antibiotics from strong resistance through partial resistance to complete sensitivity. Based on this data the mutants showing strong antibiotic resistance in vitro might be simply classified as mitochondrial ribosome mutants and mutants sensitive in vitro as mitochondrial membrane mutants; however mutants showing partial resistances are not so readily accommodated in either class. The diverse biochemical properties cannot be correlated with the different genetic loci described; indeed three mutations, each resulting in different biochemical behaviour appear to occur at the same locus. The results are interpreted as providing further evidence for an earlier proposal of mitochondrial membrane-ribosome interactions.
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Apirion, D., Schlessinger, D.: Functional interdependence of ribosomal components of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 63, 794 (1969).
Avner, P. R., Griffiths, D. E.: Oligomycin resistant mutants in yeast. FEBS Letters 10 202 (1970).
Bolotin, M., Coen, D., Deutsch, J., Dujon, B., Netter, P., Petrochilo, E., Slonimski, P. P.: La recombinaison des mitochondries chez Saccharomyces cerevisiae. Bull. Institut Pasteur 69, 215 (1971).
Bunn, C. L., Mitchell, C. H., Lukins, H. B., Linnane, A. W.: Biogenesis of mitochondria, XVIII. A new class of cytoplasmically determined antibiotic resistant mutants in Saccharomyces cerevisiae. Proc. nat. Acad. Sci. (Wash.) 67, 1233 (1970).
Clark-Walker, G. D., Linnane, A. W.: In Vivo differentiation of yeast cytoplasmic and mitochondrial protein synthesis with antibiotics. Biochem. biophys. Res. Commun. 25, 8 (1966).
Coen, D., Deutsch, J., Netter, P., Petrochilo, E., Slonimski, P. P.: Mitochondrial genetics: I. Methodology and phenomenology. In Symposium Soc. Exptl. Biol. XXIV, p. 449. London: Camb. Univ. Press 1970.
Gingold, E. B., Saunders, G. W., Lukins, H. B., Linnane, A. W.: Biogenesis of mitochondria, X Reassortment of the cytoplasmic genetic determinants for respiratory competence and erythromycin resistance in Saccharomyces cerevisiae. Genetics 62, 735 (1969).
Grivell, L. A., Reijnders, L., de Vries, H.: Altered mitochondrial ribosomes in a cytoplasmic mutant of yeast. FEBS Letters 16, 159 (1971).
Howell, N., Trembath, M. K., Linnane, A. W., Lukins, H. B.: In preparation (1973).
Huang, M., Biggs, D. R., Clark-Walker, G. D., Linnane, A. W.: Chloramphenicol inhibition of the formation of particulate mitochondrial enzymes of Saccharomyces cerevisiae. Biochim. biophys. Acta (Amst.) 114, 434 (1966).
Lai, C. J., Weisblum, B.: Altered methylation of ribosomal RNA in an erythromycin resistant strain of Staphylococcus aureus. Proc. nat. Acad. Sci. (Wash.) 68, 856 (1971).
Lamb, A. J., Clark-Walker, G. D., Linnane, A. W.: Biógenesis of mitochondria, IV. The differentiation of mitochondrial and cytoplasmic protein synthesizing systems in vitro by antibiotics. Biochim. biophys. Acta (Amst) 161, 415 (1968).
Linnane, A. W., Haslam, J. M., Lukins, H. B., Nagley, P.: The biogenesis of mitochondria in microorganisms. Ann. Rev. Microbiol. 26, 163 (1972).
Linnane, A. W., Lamb, A. J., Christodoulou, C., Lukins, H. B.: Biogenesis of mitochondria, VI. Biochemical basis of the resistance of Saccharomyces cerevisiae toward antibiotics which specifically inhibit mitochondrial protein synthesis. Proc. nat. Acad. Sci. (Wash.) 59, 1288 (1968a).
Linnane, A. W., Saunders, G. W., Gingold, E. B., Lukins, H. B.: Biogenesis of mitochondria, V. Cytoplasmic inheritance of erythromycin resistance in Saccharomyces cerevisiae. Proc. nat. Acad. Sci. (Wash.) 59, 903 (1968b).
Mitchell, C. H., Bunn, C. L., Lukins, H. B., Linnane, A. W.: The Biogenesis of mitochondria 23. The biochemical genetic characteristics of two different cytoplasmic genetically determined oligomycin resistant mutants of Saccharomyces cerevisiae. J. Bioenergetics (in press).
Saunders, G. W., Gingold, E. B., Trembath, M. K., Lukins, H. B., Linnane, A. W.: Biogenesis of mitochondria, XV. Segregation in yeast of a cytoplasmic determinant in crosses and its loss or retention in the petite. In: Autonomy and biogenesis of mitochondria and chloroplasts, p. 185. eds. N. K. Boardman and A. W. Linnane. Amsterdam: North Holland Press 1970.
Stuart, K. D.: Cytoplasmic inheritance of oligomycin and rutamycin resistance in yeast. Biochem. biophys. Res. Commun. 39, 1045 (1970).
Tanaka, K., Teraoka, H., Tamaki, M., Otaka, E., Osawa, S.: Erythromycin resistant mutant of Escherichia coli with altered ribosomal protein component. Science 162, 576 (1968).
Thomas, D. Y., Wilkie, D.: Inhibition of mitochondrial synthesis by erythromycin: Cytoplasmic and nuclear factors controlling resistance. Genet. Res. 11, 33 (1968).
Wakabayashi, K., Gunge, N.: Extrachromosomal inheritance of oligomycin resistance in yeast. FEBS Letters 6, 302 (1970).
Weisblum, B., Davies, J.: Antibiotic inhibitors of the bacterial ribosome. Bact. Rev. 32, 493 (1968).
Wilkie, D.: Analysis of mitochondrial drug resistance in Saccharomyces cerevisiae. In Symposium Soc. Exptl. Biol. XXIV. London: Camb. Univ. Press 1970.
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Trembath, M.K., Bunn, C.L., Lukins, H.B. et al. Biogenesis of mitochondria 27. Molec. gen. Genet. 121, 35–48 (1973). https://doi.org/10.1007/BF00353691
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DOI: https://doi.org/10.1007/BF00353691