Transfer Ribonucleic Acids in the Developing Brain: The Effect of the Convulsant Methionine Sulfoximine on tRNA Function

  • Orchid Der
  • Otto Z. Sellinger


Effects of the convulsant L-methionine-dl-sulfoximine (MSO) on brain transfer ribonucleic acids (tRNAs) were discovered through a comparative biochemical analysis of tRNAs from control and MSO-treated (150 mg/kg, single dose) 3-day old rats. The study of the in vitro formation of aminoacyl-tRNA, the key intermediate in translation, coupled with benzoylated-DEAE cellulose (BCD) chromatography of control and MSO-tRNAs, revealed a relationship between the molecular structure of tRNA, as affected by MSO, and its function in amino acid acceptance. Post-charging of tRNA following BDC chromatography revealed 3 tRNAphe and 3 tRNAlys isoacceptors in control brains. The administration of MSO resulted in a significant alteration of the mobility of all 6 isoacceptors on BDC and in a marked reduction of their aminoacylating capacity when compared to control molecules. The MSO-elicited impairment of aminoacyl-tRNA formation was studied using both liver (heterologous) and brain (homologous) aminoacyl-tRNA synthetase preparations. Acceptance of lysine and phenylalanine by MSOtRNA was uniformly lower than acceptance by control tRNA and was reflected by a drop in the Vmax values for the two amino acids. Different optima for[K+] and [CTP] were also found when phenylalanine acceptance by control and MSO-tRNA was compared.

The possibility that changes in brain tRNA methylation (6, 29), previously noted after MSO, may account for their impaired performance in aminoacylation is briefly discussed.


Column Buffer Develop Brain Chronic Ethanol Ingestion Methionine Sulfoximine Leucyl tRNA Synthetase 
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  1. 1.
    Barra, H.S., Unates, L.E., Sayavedra, M.S. and Caputto, R. (1972): Capacities for binding amino acids by tRNAs from rat brain and their changes during development. J. Neurochem. 19: 2289–2298.PubMedCrossRefGoogle Scholar
  2. 2.
    Carpousis, A., Christner, P. and Rosenbloom, J. (1977): Preferential usage of glycyl-tRNA isoaccepting species in collagen synthesis. J. Biol. Chem. 252: 2447–2449.Google Scholar
  3. 3.
    Christner, P.J. and Rosenbloom, J. (1976): A comparison of transfer RNA isoaccepting species between collagenous and noncollagenous tissues in the embryonic chick. Arch. Biochem. Biophys. 172: 399–409.Google Scholar
  4. 4.
    Cummins, C.J., Salas, C.E. and Sellinger, O.Z. (1975): The homologous methylation of tRNA in rat brain. Brain Res. 96: 407–412.PubMedCrossRefGoogle Scholar
  5. 5.
    Dainat, J., de Balbian Verster, F., Zand, R. and. Sellinger, O.Z. (1979): Age-dependent changes in the specificity of tRNA methyltransferases in the cerebellum of the icteric and nonicteric Gunn rat. Neurochem. Res. 4: 557–565.Google Scholar
  6. 6.
    Dainat, J., Salas, C.E., and Sellinger, O.Z. (1978): Alteration of the specificity of brain tRNA methyltransferases and of the pattern of brain tRNA methylation in vivo by methionine sulfoximine. Biochem. Pharmacol. 27: 2655–2658.Google Scholar
  7. 7.
    Dainat, J. and Sellinger, O.Z. Cerebellar tRNA methyltransferases: a developmental study. Brain Res. (in press)Google Scholar
  8. 8.
    Datta, R.K. and Antopol, W. (1973): Effect of chronic administration of morphine on mouse brain aminoacyl-tRNA synthetase and tR*TA-amino acid binding. Brain Res. 53: 373–386.PubMedCrossRefGoogle Scholar
  9. 9.
    Der, O. and Sellinggr O.Z. (19799): Changes in methylation in tRNA y and tRNAphe isoacceptor populations in the methionine sulfoximine epileptogenic rat brain. Abstracts, VII Intern. Meeting, International Soc. Neurochem., Jerusalem.Google Scholar
  10. 10.
    Drabkin, H.F. and Lukens, L.N. (1978): Prefer- ential use in collagen synthesis of the same glycyl-tRNA species that is elevated in collagen-synthesizing tissues. J. Biol. Chem. 253: 6233–6241.Google Scholar
  11. 11.
    Elahi, E. and Sellinger, O.Z. (1979): The post- natal methylation of transfer ribonucleic acid in brain. Evidence for the methylation of precursor transfer ribonucleic acid. Biochem. J. 177: 381–384.Google Scholar
  12. 12.
    Fleming, E.W., Tewari, S. and Noble, E.P. (1975): Effects of chronic ethanol ingestion on brain aminoacyl-tRNA synthetase and tRNA. J. Neurochem. 24: 553–560.PubMedCrossRefGoogle Scholar
  13. 13.
    Ginzburg, I., Cornelis, P., Giveon, D. and Littauer, U.Z. (1979): Functionally impaired tRNA. from ethionine treated rats as detected in injected Xenopus oocytes. Nucl. Acids Res. 6: 657–672.Google Scholar
  14. 14.
    Harris, C.L. and Maas, J.W. (1974): Transfer RNA and the regulation of protein synthesis in rat cerebral cortex during neural development. J. Neurochem. 22: 741–750.PubMedCrossRefGoogle Scholar
  15. 15.
    Harris, I.S. and Randerath, K. (1978): Aminoacylation of undermethylated mammalian transfer RNA. Biochim. Biophys. Acta 521: 566–575.Google Scholar
  16. 16.
    Jank, P. and Gross, H. (1974): Methyl-deficient mammalian transfer RNA: II. Homologous methylation in vitro of liver tRNA from normal and ethionine-fed rats: ethionine effect on 5-methylcytidine synthesis in vivo. Nucl. Acids. Res. 1: 1259–1267.Google Scholar
  17. 17.
    Johnson, T.C. (1969): Aminoacyl-tRNA synthetase and transfer RNA binding activity during early mammalian brain development. J. Neurochem. 16: 1125–1132.PubMedCrossRefGoogle Scholar
  18. 18.
    Johnson, T.C. (1976): Regulation of protein synthesis during postnatal maturation of the brain. J. Neurochem. 27: 17–23.PubMedCrossRefGoogle Scholar
  19. 19.
    Johnson, T.C. and Chou, L. (1972): Level and amino acid acceptor activity of mouse brain tRNA during neural development. J. Neurochem. 20: 405–414.CrossRefGoogle Scholar
  20. 20.
    Kaplan, B.B. and Sirlin, J.L. (1975): Macromolecules and behavior. II. Training induced alteration in leucine transfer RNA of goldfish brain. Brain Res. 83: 451–468.Google Scholar
  21. 21.
    Klee, H.J., DiPietro, D., Fournier, J.J. and Fischer, M.S. (1978): Characterization of transfer RNA from liver of the developing amphibian, Rana catesbeiana. J. Biol. Chem. 253: 8074–8080.Google Scholar
  22. 22.
    Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1961): Protein measurement with Folin phenol reagent. J. Biol. Chem. 193: 265–275.Google Scholar
  23. 23.
    Ofengand, J. (1977): tRNA and aminoacyl-tRNA synthetases. Molecular mechanisms of protein biosynthesis. (H. Weissbach and S. Pestka, eds.) Academic Press, New York, pp. 7–79.Google Scholar
  24. 24.
    Ortwerth, B.J. and Der, O.C. (1974): Studies on the specialized transfer RNA population of the lens. Exp. Eye Res. 19: 521–532.Google Scholar
  25. 25.
    Ortwerth, B.J., Yonuschot, G.R., Heidlege, J.F., Chu-Der, O.M.Y., Juarez, D. and Hedgcoth, C. (1975): Induction of new species of phenylalanine transfer RNA during lens cell differentiation. Exp. Eye Res. 20: 417–426.Google Scholar
  26. 26.
    Ramberg, E.S., Ishaq, M., Rulf, S., Moeller, B. and Horowitz, J. (1978): Inhibition of transfer RNA function by replacement of uridine and uridine-derived nucleosides with 5-fluorouridine. Biochemistry 17: 3878–3885.CrossRefGoogle Scholar
  27. 27.
    Roy, K.L., Bloom, A. and Soll, D. (1971): tRNA separations using benzoylated DEAE-cellulose. Procedures in Nucleic Acid Research (G.L. Cantoni and D.R. Davis, eds.) Harper and Row, New York, pp. 524–541.Google Scholar
  28. 28.
    Salas, C.E., Cummins, C.J. and Sellinger, O.Z. (1976): The developmental pattern of homologous and heterologous tRNA methylation in rat brain: differential effects of spermidine. Neurochem. Res. 1: 369–384.Google Scholar
  29. 29.
    Salas, C.E., Ohlsson, W.G. and Sellinger, O.Z. (1977): The stimulation of cerebral N2-methyl and Ni-dimethyl guanine-specific tRNA methyltransferases by methionine sulfoximine: an in vivo study. Biochem Biophys. Res. Commun. 76: 1107–1115.Google Scholar
  30. 30.
    Schimmel, P.R. (1979): Recent results on how aminoacyl transfer RNA synthetases recognize specific transfer RNAs. Molecular & Cell. Biochem. 25: 3–14.Google Scholar
  31. 31.
    Sellinger, O.Z., and Salas, C.E. (1979): Transfer RNAs in brain. Biochemistry of Brain (S. Kumar, ed.) Pergamon Press, pp. 279–297.Google Scholar
  32. 32.
    Wittig, B., Reuter, S. and Gottschling, H. (1977): Comparative characterization of four purified lysine-specific transfer ribonucleic acids from chicken embryos. J. Biochem. 81: 1705–1713.PubMedGoogle Scholar
  33. 33.
    Yang, W.K. and Novelli, G.D. (1971): Analysis of isoaccepting tRNAs in mammalian tissues and cells. Methods in Enzymology, vol. XX, part C (K. Moldave and L. Grossman, eds.) Academic Press, New York, pp. 44–55.Google Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Orchid Der
    • 1
  • Otto Z. Sellinger
    • 1
  1. 1.Laboratory of Neurochemistry Mental Health Research InstituteUniversity of Michigan Medical CenterAnn ArborUSA

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