Summary
2,6-diaminpurine (DAP) selectively inhibited mitochondrial protein synthesis in yeast cells with concomitant failure of cells to grow in non-fermentable (yeast extract, glycerol) medium. The selectivity was pronounced in all strains tested (15) nearly all of which were able to grow in yeast extract, glucose medium containing 5 mg/ml DAP (maximum solubility) whereas growth was arrested in all strains at 250–500 μg/ml DAP in the glycerol medium. The inhibition was reversed by further addition of adenine to the culture medium. RNA synthesis in rat liver mitochondria was depressed by DAP suggesting that the analogue affected RNA polymerase activity.
There was no evidence of nuclear mutagenicity by DAP but resistance to the antibiotics chloramphenicol and oligomycin was induced by the drug. Genetic evidence, although limited, indicated that the resistance mutations were cytoplasmic. The mitochondrial petite mutation was also induced by DAP but only at comparatively high concentrations. The mutagenic effects were seen only in the glycerol medium.
Similar content being viewed by others
References
Balduzzi, P.C., Morgan, H.: Effect of purine analogues on encephalomyocarditis and vaccinia virus growth in analogue sensitive and resistant cells. Proc. Soc. Exp. Biol. Med. 115, 145–148 (1964)
Benzer, S.: On the topography of the genetic fine structure. Proc. Natl. Acad. Sci. U.S.A. 47, 403–415 (1961)
Bisele, J.J., Berger, R.E., Wilson, A.Y., Hitchings, G.H., Elion, G.B.: Studies on 2,6-diaminopurine and related substances in cultures of embryonic and sarcomatous rodent tissues. Cancer 4, 186–197 (1951)
Blair, D.G.R., Dommasch, M.: Formation of inosine triphosphate and 14C-labelled 2,6-diaminopurine ribonucleoside di- and triphosphates in stored human erythrocytes. Transfusion 9, 198–202 (1969)
Burchenal, J.H., Bendich, A., Brown, G.B., Elion, G.B., Hitchings, G.H., Rhoads, C.P., Stock, C.C.: Preliminary studies on the effects of 2,6-diaminopurine on transplanted mouse leukaemia. Cancer 2, 119–120 (1949)
Burchenal, J.H., Karnofsky, D.A., Kingsley-Pillers, E.M., Southam, C.M., Myers, W.P.L., Escher, G.C., Craver, L.F., Dargeon, H.W., Rhoads, C.P.: The effects of the folic acid antagonists and 2,6-diaminopurine on neoplastic disease. Cancer 4, 549–569 (1951)
Chappell J.B., Hansford, R.G.: Preparation of mitochrondria from animal tissues and yeasts. In: Subcellular components, preparation and fractionation: (Birnie, G.D., ed.), 2nd ed. pp. 77–91. London: Butterworths: Baltimore: University Park Press 1972
Elion, G.B., Singer, S., Hitchings, G.H.: Effects of purine antagonists on a diaminopurine-resistant strain of Lactobacillus casei. J. Biol. Chem. 202, 647–654 (1953)
Faye, G., Fukuhara, H., Grandchamp, C., Lazowska, J., Michel, F., Casey, J., Getz, G.S., Locker, J., Rabinwitz, M., Bolotin-Fukuhara, M., Coen, D., Deutsch, J., Dujon, B., Netter, P., Slonimski, P.P.: Mitochondrial nucleic acids in the, petite colonie' mutants: deletions and repetitions of genes. Biochemie 55, 779–792 (1974)
Freese, B.: The specific mutagenic effect of base analogues on Phage T4. J. Mol. Biol. 1, 87–105 (1959)
Fukuhara, H., Kujawa, C.: Selective inhibition of the in vivo transcription of mitochondrial DNA by ethidium bromide and by acriflavine. Biochem. Biophys. Res. Commmun. 41, 1002–1008 (1970)
Goldthwaite, C.D., Cryer, D.R., Marmur, J.: Effect of carbon source on the replication and transmission of yeast mitochondrial mutants. Mol. Gen. Genet. 133, 87–104 (1974)
Groot, G.S.P., Van Harten-Loosbroek, N.: The use of isolated mitochondria in the study of the mitochondrial biogenesis in yeast. In Genetics, biogenesis and bioenorgetics of mitochondria (Bandlow, W., ed.), pp. 269–277. Berlin and New York: Walter de Gruyter 1976
Hartenstein, R.C., Fridovich, I.: Adenine aminohydrolase. An investigation of specificity. J. Biol. Chem. 242, 740–746 (1967)
Hitchings, G.H., Elion, G.B.: Purine analogues. In: Metabolic inhibitor (Hochster, R.M., Quastel, J.H., eds.), Vol. I, pp. 215–238. New York and London: Academic Press 1963
Johnson, R.C., Harris, V.G.: Purine analogue sensitivity and lipase activity of leptospires. Appl. Microbiol. 16, 1584–1590 (1968)
Kalle, G.P., Gots, J.S.: Genetic alteration of adenylic pyrophosphorylase in Salmonella. Science 142, 680–681 (1963)
Kaplan, H.S., Smith, K.C., Tomlin, P.: Radiosensitisation of E. coli by purine and pyrimidine analogues incorporated in deoxyribonucleic acid. Nature 190, 794–796 (1961)
Kerr, S.E., Chernigoy, F.: On the biosynthesis of ribonucleic acid purines and their interconversion in yeast. J. Biol. Chem. 200, 887–894 (1953)
Kobiler, D., Allweiss, D.: The prevention of long-term memory formation by 2,6-diaminopurine. Pharmacol. Biochem. Behav. 2, 9–17 (1974)
Kornberg, A., Pricer, W.E.: Enzymatic phosphorylation of adenosine and 2,6-diaminopurine riboside. J. Biol. Chem. 193, 481–495 (1951)
Kroon, A.M., Botman, M.J., Saccone, C.: Practical procedures for the isolation of mitochondrial preparations suitable for the study of mitochondrial macromolecules and with minimal contamination by other cell fractions or bacteria. In: Biochemical aspects of the biogenesis of mitochondria (Slater, E.C., Tager, J.M., Papa, S., Quagliariello, E., eds.), pp. 439–455. Bari: Adriatica Editrice 1968
Lee, B., Wilkie, D.: Inhibition of mitochondria of yeast by the adenine analogue 2,6-diaminopurine. Heredity 32, 282 (1974)
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J., Rose, J.: Protein measurement with the folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Luria, S.E., Delbrück, M.: Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28, 491–511 (1943)
Mortimer, R.K., Hawthorne, D.C.: Yeast genetics. The yeasts. In: (Rose, A.H., Harrison, J.S., eds.), Vol. I, pp. 386–453. London: Academic Press 1969
Putrament, A., Baranowska, H. and Prazmo, W.: Induction by manganase of mitochondrial antibiotic resistance mutations in yeast. Mol. Gen. Genet. 126, 357–366 (1973)
Rappaport, H., DeMars, R.: Diaminopurine-resistant mutants of cultured, diploid human fibroblasts. Genetics 75, 335–345 (1973)
Scragg, A.H.: A mitochondrial DNA-directed RNA polymerase from yeast mitochondria. In: The biogenesis of mitochondria Kroon, A.M., Saccone, C., eds., pp. 47–57. New York and London: Academic Press Inc. 1974
Tatibane, M., Yoshikawa, H.: Formation of 2-azaadenine and 2,6-diaminopurine analogues of adenosine triphosphaste in human erythrocytes. Biochim. Biophys. Acta 57, 613–615 (1962)
Wainfan, E., Landsberg, B.: Inhibition of transfer ribonucleic acid methylating enzymes by cytoxic analogues of adenosine. Biochem. Pharmacol. 22, 493–500 (1973)
Wheeler, G.P., Bowden, P.J.: Identification of analogues of nicotinamide adenine dinucleotide among the metabolites of 2,6-diaminopurine in mammalian cells. J. Biol. Chem. 241, 1114–1121 (1966)
Wickerham, L.J.: A critical evaluation of the nitrogen assimilation tests commonly used in the classification of yeasts. J. Bacteriol. 52, 293–301 (1946)
Wilkie, D.: Selective inhibition of mitochondrial synthesis in Saccharomyces cerevisiae by cannavanine. J. Mol. Biol. 47, 107–113 (1971)
Wilkie, D.: The yeast cell in anti-mitochondrial activity of drugs. Med. Biol. Illus. 22, 119–124 (1972)
Wilkie, D.: Cytoplasmic inheritance and mitochondrial genetics in yeast. In: Methods in cell biology (Prescott, D., ed.), Vol. XII, pp. 353–372. New York and London: Academic Press Inc. 1975
Author information
Authors and Affiliations
Additional information
Communicated by F. Kaudewitz
Rights and permissions
About this article
Cite this article
Wallis, C., Wilkie, D. Mitochondrial activity of 2,6-diaminopurine in Saccharomyces cerevisiae . Molec. Gen. Genet. 173, 307–313 (1979). https://doi.org/10.1007/BF00268641
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00268641