Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1978 Dec 1;175(3):833–840. doi: 10.1042/bj1750833

Cell-free synthesis of metallothionein directed by rat liver polyadenylated messenger ribonucleic acid.

S G Shapiro, K S Squibb, L A Markowitz, R J Cousins
PMCID: PMC1186144  PMID: 743236

Abstract

Total RNA was isolated from rat liver polyribosomes and fractionated by oligo(dT)-cellulose chromatography to obtain polyadenylated mRNA. The mRNA was translated in a wheat-germ cell-free protein-synthesizing system containing [3H]glycine, [3H]lysine and [3H]serine. Most of the newly synthesized 3H-labelled polypeptides were removed from the cell-free products by precipitation at pH 4.0. 3H-labelled thionein chains, which were soluble at pH 4.0, were purified by activated-thiol-Sepharose 4B chromatography or by gel-filtration chromatography. Polyribosomal thionein mRNA was found to increase by at least 3-fold after parenteral administration and by 20 h thereafter the ratio of thionein mRNA to total mRNA approached that found in controls. Actinomycin D administration in vivo blocked the Zn2+-induced increase in polyribosomal thionein mRNA content. These data strongly suggest that metallothionein is an inducible protein. The mechanism of regulation appears to involve changes in the synthesis de novo of thionein mRNA and hence the pool of thionein mRNA available for translation.

Full text

PDF
833

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bremner I., Davies N. T. The induction of metallothionein in rat liver by zinc injection and restriction of food intake. Biochem J. 1975 Sep;149(3):733–738. doi: 10.1042/bj1490733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bryan S. E., Hidalgo H. A. Nuclear 115cadmium: uptake and disappearance correlated with cadmium-binding protein synthesis. Biochem Biophys Res Commun. 1976 Feb 9;68(3):858–866. doi: 10.1016/0006-291x(76)91224-9. [DOI] [PubMed] [Google Scholar]
  3. Drysdale J. W., Munro H. N. Regulation of synthesis and turnover of ferritin in rat liver. J Biol Chem. 1966 Aug 10;241(15):3630–3637. [PubMed] [Google Scholar]
  4. Emtage J. S., Lawson D. E., Kodicek E. The response of the small intestine to vitamin D. Isolation and properties of chick intestinal polyribosomes. Biochem J. 1974 May;140(2):239–247. doi: 10.1042/bj1400239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Failla M. L., Cousins R. J. Zinc uptake by isolated rat liver parenchymal cells. Biochim Biophys Acta. 1978 Feb 1;538(3):435–444. doi: 10.1016/0304-4165(78)90405-1. [DOI] [PubMed] [Google Scholar]
  6. Feldman S. L., Cousins R. J. Degradation of hepatic zinc-thionein after parenteral zinc administration. Biochem J. 1976 Dec 15;160(3):583–588. doi: 10.1042/bj1600583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Green M., Zehavi-Willner T., Graves P. N., McInnes J., Pestka S. Isolation and cell-free translation of immunoglobulin messenger RNA. Arch Biochem Biophys. 1976 Jan;172(1):74–89. doi: 10.1016/0003-9861(76)90049-7. [DOI] [PubMed] [Google Scholar]
  8. Kemper B., Habener J. F., Mulligan R. C., Potts J. T., Jr, Rich A. Pre-proparathyroid hormone: a direct translation product of parathyroid messenger RNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3731–3735. doi: 10.1073/pnas.71.9.3731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Maurer R. A., Stone R., Gorski J. Cell-free synthesis of a large translation product of prolactin messenger RNA. J Biol Chem. 1976 May 10;251(9):2801–2807. [PubMed] [Google Scholar]
  10. Munro H. N. Iron absorption and nutrition: introduction. Fed Proc. 1977 Jun;36(7):2015–2016. [PubMed] [Google Scholar]
  11. Nickol J. M., Lee K. L., Hollinger T. G., Kenney F. T. Translation of messenger RNA specific for tyrosine aminotransferase in oocytes of Xenopus laevis. Biochem Biophys Res Commun. 1976 Sep 20;72(2):687–693. doi: 10.1016/s0006-291x(76)80094-0. [DOI] [PubMed] [Google Scholar]
  12. Palmiter R. D. Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undergraded polysomes and messenger ribonucleic acid. Biochemistry. 1974 Aug 13;13(17):3606–3615. doi: 10.1021/bi00714a032. [DOI] [PubMed] [Google Scholar]
  13. Prosky L., Roberts B., Jr, O'Dell R. G., Imblum R. L. Differential effects of actinomycin D on nucleic acid and protein synthesis in rat liver. Arch Biochem Biophys. 1968 Aug;126(2):393–398. doi: 10.1016/0003-9861(68)90423-2. [DOI] [PubMed] [Google Scholar]
  14. Richards M. P., Cousins R. J. Influence of inhibitors of protein synthesis on zinc metabolism. Proc Soc Exp Biol Med. 1977 Dec;156(3):505–508. doi: 10.3181/00379727-156-39967. [DOI] [PubMed] [Google Scholar]
  15. Richards M. P., Cousins R. J. Mammalian zinc homeostasis: requirement for RNA and metallothionein synthesis. Biochem Biophys Res Commun. 1975 Jun 16;64(4):1215–1223. doi: 10.1016/0006-291x(75)90822-0. [DOI] [PubMed] [Google Scholar]
  16. Richards M. P., Cousins R. J. Zinc-binding protein: relationship to short term changes in zinc metabolism. Proc Soc Exp Biol Med. 1976 Oct;153(1):52–56. doi: 10.3181/00379727-153-39479. [DOI] [PubMed] [Google Scholar]
  17. Roewekamp W. G., Hofer E., Sekeris C. E. Translation of mRNA from rat-liver polysomes into tyrosine aminotransferase and tryptophan oxygenase in a protein-synthesizing system from wheat germ. Effects of cortisol on the translatable levels of mRNA for these two enzymes. Eur J Biochem. 1976 Nov 1;70(1):259–268. doi: 10.1111/j.1432-1033.1976.tb10977.x. [DOI] [PubMed] [Google Scholar]
  18. Schutz G., Beato M., Feigelson P. Messenger RNA for hepatic tryptophan oxygenase: its partial purification, its translation in a heterologous cell-free system, and its control by glucocorticoid hormones. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1218–1221. doi: 10.1073/pnas.70.4.1218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Spencer R., Charman M., Lawson D. E., Emtage J. S. Production and properties of vitamin-D-induced mRNA for chick calcium-binding protein. Eur J Biochem. 1976 Dec 11;71(2):399–409. doi: 10.1111/j.1432-1033.1976.tb11127.x. [DOI] [PubMed] [Google Scholar]
  20. Squibb K. S., Cousins R. J., Feldman S. L. Control of zinc-thionein synthesis in rat liver. Biochem J. 1977 Apr 15;164(1):223–228. doi: 10.1042/bj1640223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Squibb K. S., Cousins R. J. Synthesis of metallothionein in a polysomal cell-free system. Biochem Biophys Res Commun. 1977 Apr 11;75(3):806–812. doi: 10.1016/0006-291x(77)91544-3. [DOI] [PubMed] [Google Scholar]
  22. Weser U., Bischoff E. Incorporation of 65Zn in rat liver nuclei. Eur J Biochem. 1970 Feb;12(3):571–575. doi: 10.1111/j.1432-1033.1970.tb00888.x. [DOI] [PubMed] [Google Scholar]
  23. Weser U., Hübner L. Cd(2+), Mn(2+) and Zn(2+) induced synthesis of nuclear RNA in the livers of normal and adrenalectomized rats. FEBS Lett. 1970 Oct 5;10(3):169–174. doi: 10.1016/0014-5793(70)80445-8. [DOI] [PubMed] [Google Scholar]
  24. Weser U., Hübner L., Jung H. Zn(2+)-Induced stimulation of nuclear RNA synthesis in rat liver. FEBS Lett. 1970 May 1;7(4):356–358. doi: 10.1016/0014-5793(70)80205-8. [DOI] [PubMed] [Google Scholar]
  25. Zähringer J., Baliga B. S., Munro H. N. Novel mechanism for translational control in regulation of ferritin synthesis by iron. Proc Natl Acad Sci U S A. 1976 Mar;73(3):857–861. doi: 10.1073/pnas.73.3.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zähringer J., Konijn A. M., Baliga B. S., Munro H. N. Mechanism of iron induction of ferritin synthesis. Biochem Biophys Res Commun. 1975 Jul 22;65(2):583–590. doi: 10.1016/s0006-291x(75)80186-0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES