Summary
Cytoplasmic petite mutagenesis by 5-fluorouracil (5FU) was prevented by temperature sensitive mutations which blocked either nuclear transcription or cytoplasmic translation. However, 5FU was also ineffective in resting cells and in cells exposed to α-mating factor, showing that cell division or nuclear DNA synthesis is required for the mutagenic event to take place. In addition, the mutagenic effect of 5FU was completely prevented by daunomycin, and since this agent preferentially inhibits respiratory growth and was shown to selectively block RNA synthesis in the mitochondria, it was concluded that petite mutagenesis resulted from incorporation of 5FU into mitochondrial RNA. Since inhibition of mitochondrial protein synthesis by erythromycin had little immediate effect on the mutagenicity of 5FU, it was deduced that the RNA in question is not directly involved in mitochondrial translation, and may have a regulatory function.
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References
Beech, F.W., Carr, J.G., Codner, R.C.: A multi-point inoculator for plating bacteria or yeasts. J. gen. Microbiol.13, 408–410 (1955)
Bevan, E.A., Herring, A.J., Mitchell, D.J.: Two species of dsRNA in yeast: their preliminary characterization and relationship to the killer character. Nature (Lond.)245, 81–86 (1973)
Bray, G.A.: A simple and efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analyt. Biochem.1, 279–285 (1960)
Bucking-Throm, E., Duntze, W., Hartwell, L.H., Manney, T.R.: Reversible arrest of haploid yeast cells at the initiation of DNA synthesis by a diffusible sex factor. Exp. Cell Res.76, 99–110 (1973)
Eaton, N.R.: New press for the disruption of microorganisms. J. Bact.83, 1359–1360 (1962)
Evans, I., Linstead, D., Rhodes, P.M., Wilkie, D.: Inhibition of RNA synthesis in mitochondria by daunomycin. Biochim. biophys. Acta (Amst.)312, 323–336 (1973)
Hartwell, L.H., McLaughlin, C.S.: A mutant apparently defective in the initiation of protein synthesis. Proc. nat. Acad. Sci. (Wash.)62, 468–474 (1969)
Hutchinson, H.T., Hartwell, L.H., McLaughlin, C.S.: A temperature-sensitive yeast mutant defective in RNA production. J. Bact.99, 807–814 (1969)
Kirby, K.S.: Isolation and characterization of ribosomal ribonucleic acid. Biochem. J.96, 266–269 (1965)
Kraepelin, G.: Some viewpoints concerning respiratory deficiency in Yeast. In Yeast: Proceedings 2nd Symposium on Yeasts, Bratislava, p. 237. Prague: Czechoslovak Academy of Sciences 1969
Lamb, A.J., Clark-Walker, G.D., Linnane, A.W.: The biogenesis of mitochondria 4: The differentiation of mitochondrial and cytoplasmic protein synthesizing systemsin vitro by antibiotics. Biochim. biophys. Acta (Amst.)161, 415–427 (1968)
Loening, U.E.: The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide gel electrophoresis. Biochem. J.102, 251–257 (1967)
Mahler, H.R., Dawidowicz, K.: Autonomy of mitochondria inSaccharomyces cerevisiae in their production of messenger RNA. Proc. nat. Acad. Sci. (Wash.)70, 111–114 (1973)
Oliver, S.G.: The role of RNA in the maintenance of mitochondrial DNA in the yeastSaccharomyces cerevisiae. Thesis CNAA (1974a)
Oliver, S.G.: Mutants of yeast specifically resistant to petite induction by 5-fluorouracil. Heredity33, 132 (1974b)
Oliver, S.G., Williamson, D.H.: The molecular events involved in the induction of petite yeast mutants by fluorinated pyrimidines. Molec. gen. Genet.146, 253–259 (1976)
Petes, T.D., Fangman, W.L.: Preferential synthesis of yeast mitochondrial DNA in α-factor arrested cells. Biochem. biophys. Res. Commun.55, 606–609 (1973)
Throm, E., Duntze, W.: Mating-type-dependent inhibition of deoxyribonucleic acid synthesis inSaccharomyces cerevisiae. J. Bact.104, 1388–1390 (1970)
Williamson, D.H., Fennell, D.J.: The use of a fluorescent DNA-binding dye for detecting and separating yeast mitochondrial DNA. Meth. Cell Biol.12, 335–351 (1975)
Williamson, D.H., Maroudas, N.G., Wilkie, D.: Induction of the cytoplasmic petite mutation inSaccharomyces cerevisiae by the antibacterial antibiotics erythromycin and chloramphenicol. Molec. gen. Genet.111, 209–223 (1971)
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Communicated by W. Gajewski
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Oliver, S.G., Williamson, D.H. The conditions required for the induction of petite yeast mutants by fluorinated pyrimidines. Molec. Gen. Genet. 146, 261–268 (1976). https://doi.org/10.1007/BF00701249
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DOI: https://doi.org/10.1007/BF00701249