Summary
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1.
Mitochondrial ribosomes have been isolated from five strains of Saccharomyces cerevisiae each carrying a mitochondrial mutation conferring in vivo resistance to spiramycin.
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2.
The response of poly U directed poly-phenylalanine synthesis to the antibiotics spiramycin, carbomycin and lincomycin was tested using the ribosomes of each mutant. The results indicate that all mutants possess altered ribosomes. Genetic mapping studies suggest that the 21S rRNA may be modified in each mutant.
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3.
The expression of antibiotic resistance by the mitochondrial ribosomes of some strains is partially or totally dependent upon an association with the mitochondrial membrane. Interpretation of the protein synthesis data is made in terms of the existence of a functional interaction between the ribosomes and the membrane in vivo.
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4.
Phenotypic interactions have been observed between different ribosomal mutations including ery1, ery2, spi2, spi3 and spi4 and a mutation affecting the mitochondrial membrane, oli. The expression of the ribosomal mutation is often markedly affected by the presence of the oli1 membrane mutation.
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5.
Membrane effects very similar to those observed following the introduction of the oli1 mutation, such as the unmasking of antibiotic sensitive ribosomal sites, can be obtained by the treatment in vivo of certain antibiotic resistant strains with non-toxic levels of the detergent SDS (< 0.01%).
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6.
The SDS effect together with the observation of phenotypic interactions between mutations provide strong supportive evidence of a functional association between a membrane component (in this case of the mtATPase) and a component of the mitochondrial ribosomes.
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References
Bunn, C.L., Mitchell, C.H., Lukins, H.B., Linnane, A.W.: Biogenesis of mitochondria 18. A new class of mitochondrial cytoplamically determined antibiotic resistance mutants in Saccharomyces cerevisiae. Proc. nat. Acad. Sci. (Wash.) 67, 1233–1240 (1971)
Ceriotti, G.: Determination of nucleic acids in animal tissues. J. biol. Chem. 214, 59–70 (1955)
Choo, K.B., Nagley, P., Lukins, H.B., Linnane, A.W.: Biogenesis of mitochondria 47. Refined physical map of the mitochondrial genome of Saccharomyces cerevisiae determined by analysis of an extended library of genetically and molecularly defined petite strains. Molec. gen. Genet. 153, 279–288 (1977)
Cobon, G.S., Crowfoot, P.D., Linnane, A.W.: Biogenesis of mitochondria 35. Phospholipid synthesis in vitro by yeast mitochondrial and microsomal fractions. Biochem. J. 144, 265–275 (1974)
Gornall, A.G., Bardawill, C.J., David, M.M.: Determination of serum proteins by means of the biuret reaction. J. biol. Chem. 177, 751–766 (1949)
Grivell, L.A., Reijnders, L., Borst, P.: Isolation of yeast mitochondrial ribosomes highly active in protein synthesis. Biochim. biophys. Acta (Amst.) 247, 91–103 (1971)
Groot Obbink, D.J., Hall, R.M., Linnane, A.W., Lukins, H.B., Monk, B.C., Spithill, T.W., Trembath, M.K.: Mitochondrial genes involved in the determination of mitochondrial membrane proteins. In: The genetic function of mitochondrial DNA. (Saccone, C. and Kroon, A.M. eds.), pp. 163–173. Amsterdam: North Holland 1976
Groot Obbink, D.J., Spithill, T.W., Maxwell, R.J., Linnane, A.W.: Biogenesis of mitochondria 48. Mikamycin resistance in Saccharomyces cerevisiae — a mitochondrial mutation conferring resistance to an antimycin A-like contaminant in mikamycin. Molec. gen. Genet. 151, 127–136 (1977)
Howell, N., Trembath, M.K., Linnane, A.W., Lukins, H.B.: Biogenesis of mitochondria 30. An analysis of polarity of mitochondrial gene recombination and transmission. Molec. gen. Genet. 122, 37–51 (1973)
Lamb, A.J., Clark-Walker, G.D., Linnane, A.W.: Biogenesis of mitochondria 4. The differentiation of mitochondrial and cytoplasmic protein synthesizing systems in vitro by antibiotics. Biochim. biophys. Acta (Amst.) 161, 415–427 (1968)
Linnane, A.W., Nagley, P., Hall, R.M., Marzuli., S., Trembath, M.K.: Molecular and genetic aspects of the expression of mitochondrial DNA in Saccharomyces cerevisiae. In: International Symposium on pure and applied biochemistry of yeasts University of Sao Paulo, Brazil (1978)
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. nat. Acad. Sci. (Wash.) 59, 903–910 (1968)
Linnane, A.W., Ward, K.A., Forrester, L.T., Haslam, J.M., Plummer, D.T.: The role of lipids in mitochondrial membrane function. In: Yeast, mould and plant protoplasts (Villaneuva, J.P., Garcia-Acha, I., Gascon, S. and Urabura, F., eds.), pp. 344–360. New York: Academic Press 1973
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the phenol reagent. J. biol. Chem. 193, 265–275 (1951)
Marzuki, S., Cobon, G.S., Haslam, J.M., Linnane, A.W.: Biogenesis of mitochondria 38. The effects of altered steady-state membrane lipid composition on mitochondrial energy metabolism in Saccharomyces cerevisiae. Arch. Biochem. Biophys. 169, 577–590 (1975)
Modolell, J.: The S30 system from Escherichia coli. In: Protein biosynthesis in bacterial systems (Last J.A. and Laskin, A.I., eds.), pp. 1–65. New York: Dekker 1971
Reijnders, L., Sloof, P., Borst, P.: The molecular weights of the mitochondrial ribosomal RNAs of Saccharomyces cerevisiae. Europ. J. Biochem. 35, 266–269 (1973)
Spithill, T.W., Nagley, P., Linnane, A.W.: Biochemical characterization of a mitochondrial mutation in Saccharomyces cerevisiae affecting the mitochondrial ribosome by conferring resistance to aminoglycoside antibiotics. (In preparation)
Trembath, M.K.: Mitochondrial genetics in yeast: Biochemical and genetic analysis of mutations conferring resistance to macrolide antibiotics. Ph.D. Thesis, Monash University, Australia (1973)
Trembath, M.K., Bunn, C.L., Lukins, H.B., Linnane, A.W.: Biogenesis of mitochondria 27. Genetic and biochemical characterization of cytoplasmic and nuclear mutations to spiramycin resistance in Saccharomyces cerevisiae. Molec. gen. Genet. 121, 35–48 (1973)
Trembath, M.K., Molloy, P.L., Sriprakash, K.S., Cutting, G.J., 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)
Trembath, M.K., Monk, B.C., Kellerman, G.M., Linnane, A.W.: Biogenesis of mitochondria 40 Phenotypic suppression of a mitochondrial mutation by a nuclear gene in Saccharomyces cerevisiae. Molec. gen. Genet. 140 333–337 (1975)
Vazquez, D.: Antibiotics effecting chloramphenicol uptake by bacteria. Their effect on amino acid incorporation in a cell free system. Biochim. biophys. Acta (Amst.) 114, 289–295 (1966)
Wallace, P.G., Huang, M., Linnane, A.W.: Biogenesis of mitochondria II. The influence of medium composition on the cytology of anaerobically grown S. cerevisiae. J. Cell Biol. 37, 207–220 (1968)
Wolf, K., Dujon, B., Slonimski, P.: Mitochondrial genetics V, Multifactorial mitochondrial crosses involving a mutation conferring paromomycin-resistance in Saccharomyces cerevisiae. Molec. gen. Genet. 125, 53–90 (1973)
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Communicated by W. Gajewski
Publication number 50 in the series Biogenesis of Mitochondria
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Spithill, T.W., Trembath, M.K., Lukins, H.B. et al. Mutations of the mitochondrial DNA of Saccharomyces cerevisiae which affect the interaction between mitochondrial ribosomes and the inner mitochondrial membrane. Molec. Gen. Genet. 164, 155–162 (1978). https://doi.org/10.1007/BF00267380
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DOI: https://doi.org/10.1007/BF00267380