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. 1985 Jan 1;225(1):77–84. doi: 10.1042/bj2250077

Pretranslational regulation of tyrosine aminotransferase and phosphoenolpyruvate carboxykinase (GTP) synthesis by glucagon and dexamethasone in adult rat hepatocytes.

P B Iynedjian, P Auberger, Y Guigoz, A Le Cam
PMCID: PMC1144555  PMID: 2858199

Abstract

The regulation of synthesis of the gluconeogenic cytosolic enzyme phosphoenolpyruvate carboxykinase (PEPCK) and of tyrosine aminotransferase (TAT) by glucagon and glucocorticoid hormones was studied in hepatocytes maintained in suspension culture for 7 h. Specific antibodies were used to measure relative rates of enzyme synthesis after pulse-labelling of the cells with [3H]leucine or [35S]methionine. Concomitantly, amounts of mRNA were quantified after translation in vitro in a reticulocyte lysate and specific immunoprecipitation of the proteins. Glucagon stimulated the rate of synthesis of PEPCK by 4-6-fold and that of TAT by 6-8-fold in 2h. In contrast, dexamethasone had little effect on PEPCK synthesis, whereas it increased TAT synthesis by 5-9-fold. When used in combination, the two hormones displayed additive effects on TAT synthesis, whereas the glucocorticoid hormone strongly potentiated stimulation of PEPCK synthesis by glucagon. In every instance, changes in rates of synthesis of the two enzymes were totally accounted for by increases in amounts of the corresponding functional mRNA, suggesting a pretranslational site of action for both glucagon and dexamethasone.

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  1. Auberger P., Samson M., Le Cam A. Inhibition of hormonal induction of tyrosine aminotransferase by polyamines in freshly isolated rat hepatocytes. Biochem J. 1983 Sep 15;214(3):679–685. doi: 10.1042/bj2140679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beale E. G., Hartley J. L., Granner D. K. N6,O2'-dibutyryl cycle AMP and glucose regulate the amount of messenger RNA coding for hepatic phosphoenolpyruvate carboxykinase (GTP). J Biol Chem. 1982 Feb 25;257(4):2022–2028. [PubMed] [Google Scholar]
  3. Chandler V. L., Maler B. A., Yamamoto K. R. DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo. Cell. 1983 Jun;33(2):489–499. doi: 10.1016/0092-8674(83)90430-0. [DOI] [PubMed] [Google Scholar]
  4. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  5. Chrapkiewicz N. B., Beale E. G., Granner D. K. Induction of the messenger ribonucleic acid coding for phosphoenolpyruvate carboxykinase in H4-II-E cells. Evidence for a nuclear effect of cyclic AMP. J Biol Chem. 1982 Dec 10;257(23):14428–14432. [PubMed] [Google Scholar]
  6. Cimbala M. A., Lamers W. H., Nelson K., Monahan J. E., Yoo-Warren H., Hanson R. W. Rapid changes in the concentration of phosphoenolpyruvate carboxykinase mRNA in rat liver and kidney. Effects of insulin and cyclic AMP. J Biol Chem. 1982 Jul 10;257(13):7629–7636. [PubMed] [Google Scholar]
  7. Culpepper J. A., Liu A. Y. Pretranslational control of tyrosine aminotransferase synthesis by 8-bromo-cyclic AMP in H-4 rat hepatoma cells. J Biol Chem. 1983 Nov 25;258(22):13812–13819. [PubMed] [Google Scholar]
  8. Diesterhaft M., Noguchi T., Granner D. Regulation of rat-liver tyrosine-aminotransferase mRNA by hydrocortisone and by N6,O2'-dibutyryladenosine 3',5'-phosphate. Eur J Biochem. 1980 Jul;108(2):357–365. doi: 10.1111/j.1432-1033.1980.tb04730.x. [DOI] [PubMed] [Google Scholar]
  9. Ernest M. J., Chen C. L., Feigelson P. Induction of tyrosine aminotransferase synthesis in isolated liver cell suspensions. Absolute dependence of induction on glucocorticoids and glucagon or cyclic AMP. J Biol Chem. 1977 Oct 10;252(19):6783–6791. [PubMed] [Google Scholar]
  10. Ernest M. J., Feigelson P. Increase in hepatic tyrosine aminotransferase mRNA during enzyme induction by N6,O2'-dibutyryl cyclic AMP. J Biol Chem. 1978 Jan 25;253(2):319–322. [PubMed] [Google Scholar]
  11. Granner D. K., Hargrove J. L. Regulation of the synthesis of tyrosine aminotransferase: the relationship to mRNATAT. Mol Cell Biochem. 1983;53-54(1-2):113–128. doi: 10.1007/BF00225249. [DOI] [PubMed] [Google Scholar]
  12. Gunn J. M., Hanson R. W., Meyuhas O., Reshef L., Ballard F. J. Glucocorticoids and the regulation of phosphoenolpyruvate carboxykinase (guanosine triphosphate) in the rat. Biochem J. 1975 Aug;150(2):195–203. doi: 10.1042/bj1500195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gurr J. A., Potter V. R. The significance of differences between fresh cell suspensions and fresh or maintained monolayers. Ann N Y Acad Sci. 1980;349:57–66. doi: 10.1111/j.1749-6632.1980.tb29514.x. [DOI] [PubMed] [Google Scholar]
  14. Hanson R. W., Fisher L., Ballard F. J., Reshef L. The regulation of phosphoenolpyruvate carboxykinase in fetal rat liver. Enzyme. 1973;15(1):97–110. [PubMed] [Google Scholar]
  15. Hargrove J. L., Diesterhaft M., Noguchi T., Granner D. K. Identification of native tyrosine aminotransferase and an explanation for the multiple forms. J Biol Chem. 1980 Jan 10;255(1):71–78. [PubMed] [Google Scholar]
  16. Hargrove J. L., Granner D. K. Physical properties, limited proteolysis, and acetylation of tyrosine aminotransferase from rat liver. J Biol Chem. 1981 Aug 10;256(15):8012–8017. [PubMed] [Google Scholar]
  17. Higgins R. C., Dahmus M. E. Rapid visualization of protein bands in preparative SDS-polyacrylamide gels. Anal Biochem. 1979 Mar;93(2):257–260. doi: 10.1016/s0003-2697(79)80148-7. [DOI] [PubMed] [Google Scholar]
  18. Iynedjian P. B., Hanson R. W. Increase in level of functional messenger RNA coding for phosphoenolpyruvate carboxykinase (GTP) during induction by cyclic adenosine 3':5'-monophosphate. J Biol Chem. 1977 Jan 25;252(2):655–662. [PubMed] [Google Scholar]
  19. Iynedjian P. B., Jacot M. M. Coordinate induction of several mRNA species in rat kidney during glucocorticoid treatment. J Biol Chem. 1981 Jul 10;256(13):7068–7076. [PubMed] [Google Scholar]
  20. Iynedjian P. B., Jacot M. M. Glucocorticoid-dependent induction of mRNA coding for phosphoenolpyruvate carboxykinase (GTP) in rat kidney. Its inhibition by cycloheximide. Eur J Biochem. 1980 Oct;111(1):89–98. doi: 10.1111/j.1432-1033.1980.tb06078.x. [DOI] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Lamers W. H., Hanson R. W., Meisner H. M. cAMP stimulates transcription of the gene for cytosolic phosphoenolpyruvate carboxykinase in rat liver nuclei. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5137–5141. doi: 10.1073/pnas.79.17.5137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Noguchi T., Diesterhaft M., Granner D. Dibutyryl cyclic AMP increases the amount of functional messenger RNA coding for tyrosine aminotransferase in rat liver. J Biol Chem. 1978 Mar 10;253(5):1332–1335. [PubMed] [Google Scholar]
  24. Noguchi T., Diesterhaft M., Granner D. Evidence for a dual effect of dibutyryl cyclic AMP on the synthesis of tyrosine aminotransferase in rat liver. J Biol Chem. 1982 Mar 10;257(5):2386–2390. [PubMed] [Google Scholar]
  25. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  26. Olson P. S., Thompson E. B., Granner D. K. Regulation of hepatoma tissue culture cell tyrosine aminotransferase messenger ribonucleic acid by dexamethasone. Biochemistry. 1980 Apr 15;19(8):1705–1711. doi: 10.1021/bi00549a029. [DOI] [PubMed] [Google Scholar]
  27. Payvar F., Wrange O., Carlstedt-Duke J., Okret S., Gustafsson J. A., Yamamoto K. R. Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6628–6632. doi: 10.1073/pnas.78.11.6628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pfahl M., McGinnis D., Hendricks M., Groner B., Hynes N. E. Correlation of glucocorticoid receptor binding sites on MMTV proviral DNA with hormone inducible transcription. Science. 1983 Dec 23;222(4630):1341–1343. doi: 10.1126/science.6318311. [DOI] [PubMed] [Google Scholar]
  29. Pfahl M. Specific binding of the glucocorticoid-receptor complex to the mouse mammary tumor proviral promoter region. Cell. 1982 Dec;31(2 Pt 1):475–482. doi: 10.1016/0092-8674(82)90140-4. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Salavert A., Iynedjian P. B. Regulation of phosphoenolpyruvate carboxykinase (GTP) synthesis in rat liver cells. Rapid induction of specific mRNA by glucagon or cyclic AMP and permissive effect of dexamethasone. J Biol Chem. 1982 Nov 25;257(22):13404–13412. [PubMed] [Google Scholar]
  32. Scherer G., Schmid W., Strange C. M., Röwekamp W., Schütz G. Isolation of cDNA clones coding for rat tyrosine aminotransferase. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7205–7208. doi: 10.1073/pnas.79.23.7205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Seglen P. O. Preparation of isolated rat liver cells. Methods Cell Biol. 1976;13:29–83. doi: 10.1016/s0091-679x(08)61797-5. [DOI] [PubMed] [Google Scholar]
  34. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  35. Wicks W. D., Kenney F. T., Lee K. L. Induction of hepatic enzyme synthesis in vivo by adenosine 3', 5'-monophosphate. J Biol Chem. 1969 Nov 10;244(21):6008–6013. [PubMed] [Google Scholar]
  36. Wicks W. D. Regulation of protein synthesis by cyclic AMP. Adv Cyclic Nucleotide Res. 1974;4(0):335–438. [PubMed] [Google Scholar]
  37. Yoo-Warren H., Monahan J. E., Short J., Short H., Bruzel A., Wynshaw-Boris A., Meisner H. M., Samols D., Hanson R. W. Isolation and characterization of the gene coding for cytosolic phosphoenolpyruvate carboxykinase (GTP) from the rat. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3656–3660. doi: 10.1073/pnas.80.12.3656. [DOI] [PMC free article] [PubMed] [Google Scholar]

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