Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1991 Sep;88(3):811–816. doi: 10.1172/JCI115381

Altered transcriptional regulation of phosphoenolpyruvate carboxykinase in rats following endotoxin treatment.

M Hill 1, R McCallum 1
PMCID: PMC295466  PMID: 1653277

Abstract

The molecular mechanism involved in altered regulation of the rate-limiting enzyme in hepatic gluconeogenesis, phosphoenolpyruvate carboxykinase (PEPCK), during endotoxemia is not completely understood. We examined, therefore, the effect of a nonlethal dose of Escherichia coli endotoxin on PEPCK gene expression in fasted rats. 5 h after endotoxin treatment, the PEPCK transcription rate and the amount of mRNA(PEPCK) were significantly decreased at a time when the insulin/glucagon (I/G) molar ratio and plasma corticosterone levels were significantly increased. Similar results were observed in a time course study, in which altered cAMP induction of PEPCK gene expression paralleled changes in the I/G molar ratio. In diabetic rats treated with endotoxin, PEPCK gene expression was decreased in the absence, however, of an increased I/G molar ratio. This finding indicates that other factors, such as inflammatory mediators or cytokines, alter PEPCK gene transcription during endotoxemia. IL-6, a putative mediator of endotoxin action in the liver, had no effect on PEPCK gene expression in fasted rats, but did decrease cAMP induction of PEPCK gene expression. These results indicate that, during endotoxemia, regulation of PEPCK gene expression is influenced by inflammatory mediators in addition to the classical endocrine hormones. IL-6, however, does not appear to be involved directly in the altered regulation of the PEPCK gene during endotoxemia.

Full text

PDF
811

Selected References

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

  1. Akira S., Hirano T., Taga T., Kishimoto T. Biology of multifunctional cytokines: IL 6 and related molecules (IL 1 and TNF). FASEB J. 1990 Aug;4(11):2860–2867. [PubMed] [Google Scholar]
  2. Baumann H., Gauldie J. Regulation of hepatic acute phase plasma protein genes by hepatocyte stimulating factors and other mediators of inflammation. Mol Biol Med. 1990 Apr;7(2):147–159. [PubMed] [Google Scholar]
  3. Beale E. G., Chrapkiewicz N. B., Scoble H. A., Metz R. J., Quick D. P., Noble R. L., Donelson J. E., Biemann K., Granner D. K. Rat hepatic cytosolic phosphoenolpyruvate carboxykinase (GTP). Structures of the protein, messenger RNA, and gene. J Biol Chem. 1985 Sep 5;260(19):10748–10760. [PubMed] [Google Scholar]
  4. 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]
  5. Beale E. G., Katzen C. S., Granner D. K. Regulation of rat liver phosphoenolpyruvate carboxykinase (GTP) messenger ribonucleic acid activity by N6, O2'-dibutyryladenosine 3',5'-phosphate. Biochemistry. 1981 Aug 18;20(17):4878–4883. doi: 10.1021/bi00520a012. [DOI] [PubMed] [Google Scholar]
  6. Berry L. J., Huff W. B. Antiglucocorticoid effects of endotoxin. Circ Shock. 1984;14(2):137–145. [PubMed] [Google Scholar]
  7. Berry L. J., Smythe D. S., Colwell L. S. Inhibition of hepatic enzyme induction as a sensitive assay for endotoxin. J Bacteriol. 1968 Oct;96(4):1191–1199. doi: 10.1128/jb.96.4.1191-1199.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang H. C., Lane M. D. The enzymatic carboxylation of phosphoenolpyruvate. II. Purification and properties of liver mitochondrial phosphoenolpyruvate carboxykinase. J Biol Chem. 1966 May 25;241(10):2413–2420. [PubMed] [Google Scholar]
  9. 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]
  10. Exton J. H. Gluconeogenesis. Metabolism. 1972 Oct;21(10):945–990. doi: 10.1016/0026-0495(72)90028-5. [DOI] [PubMed] [Google Scholar]
  11. Fey G. H., Gauldie J. The acute phase response of the liver in inflammation. Prog Liver Dis. 1990;9:89–116. [PubMed] [Google Scholar]
  12. Filkins J. P. Endotoxin-enhanced secretion of macrophage insulin-like activity. J Reticuloendothel Soc. 1980 May;27(5):507–511. [PubMed] [Google Scholar]
  13. Fong Y., Moldawer L. L., Marano M., Wei H., Tatter S. B., Clarick R. H., Santhanam U., Sherris D., May L. T., Sehgal P. B. Endotoxemia elicits increased circulating beta 2-IFN/IL-6 in man. J Immunol. 1989 Apr 1;142(7):2321–2324. [PubMed] [Google Scholar]
  14. Granner D. K. The molecular biology of insulin action on protein synthesis. Kidney Int Suppl. 1987 Dec;23:S82–S96. [PubMed] [Google Scholar]
  15. Granner D., Andreone T., Sasaki K., Beale E. Inhibition of transcription of the phosphoenolpyruvate carboxykinase gene by insulin. Nature. 1983 Oct 6;305(5934):549–551. doi: 10.1038/305549a0. [DOI] [PubMed] [Google Scholar]
  16. Granner D., Pilkis S. The genes of hepatic glucose metabolism. J Biol Chem. 1990 Jun 25;265(18):10173–10176. [PubMed] [Google Scholar]
  17. 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]
  18. Gunn J. M., Tilghman S. M., Hanson R. W., Reshef L., Ballard F. J. Effects of cyclic adenosine monophosphate, dexamethasone and insulin on phosphoenolpyruvate carboxykinase synthesis in Reuber H-35 hepatoma cells. Biochemistry. 1975 Jun 3;14(11):2350–2357. doi: 10.1021/bi00682a012. [DOI] [PubMed] [Google Scholar]
  19. Hill M. R., Stith R. D., McCallum R. E. Human recombinant IL-1 alters glucocorticoid receptor function in Reuber hepatoma cells. J Immunol. 1988 Sep 1;141(5):1522–1528. [PubMed] [Google Scholar]
  20. Hill M. R., Stith R. D., McCallum R. E. Interleukin 1: a regulatory role in glucocorticoid-regulated hepatic metabolism. J Immunol. 1986 Aug 1;137(3):858–862. [PubMed] [Google Scholar]
  21. Hill M. R., Stith R. D., McCallum R. E. Monokines mediate decreased hepatic glucocorticoid binding in endotoxemia. J Leukoc Biol. 1987 Mar;41(3):236–241. doi: 10.1002/jlb.41.3.236. [DOI] [PubMed] [Google Scholar]
  22. Hod Y., Hanson R. W. Cyclic AMP stabilizes the mRNA for phosphoenolpyruvate carboxykinase (GTP) against degradation. J Biol Chem. 1988 Jun 5;263(16):7747–7752. [PubMed] [Google Scholar]
  23. Imai E., Stromstedt P. E., Quinn P. G., Carlstedt-Duke J., Gustafsson J. A., Granner D. K. Characterization of a complex glucocorticoid response unit in the phosphoenolpyruvate carboxykinase gene. Mol Cell Biol. 1990 Sep;10(9):4712–4719. doi: 10.1128/mcb.10.9.4712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Jablons D. M., Mulé J. J., McIntosh J. K., Sehgal P. B., May L. T., Huang C. M., Rosenberg S. A., Lotze M. T. IL-6/IFN-beta-2 as a circulating hormone. Induction by cytokine administration in humans. J Immunol. 1989 Mar 1;142(5):1542–1547. [PubMed] [Google Scholar]
  26. Kioussis D., Reshef L., Cohen H., Tilghman S. M., Iynedjian P. B., Ballard F. J., Hanson R. W. Alterations in translatable messenger RNA coding for phosphoenolpyruvate carboxykinase (GTP) in rat liver cytosol during deinduction. J Biol Chem. 1978 Jun 25;253(12):4327–4332. [PubMed] [Google Scholar]
  27. Marinkovic S., Jahreis G. P., Wong G. G., Baumann H. IL-6 modulates the synthesis of a specific set of acute phase plasma proteins in vivo. J Immunol. 1989 Feb 1;142(3):808–812. [PubMed] [Google Scholar]
  28. McCallum R. E., Berry L. J. Effects of endotoxin on gluconeogenesis, glycogen synthesis, and liver glycogen synthase in mice. Infect Immun. 1973 Apr;7(4):642–654. doi: 10.1128/iai.7.4.642-654.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. McCallum R. E., Seale T. W., Stith R. D. Influence of endotoxin treatment on dexamethasone induction of hepatic phosphoenolpyruvate carboxykinase. Infect Immun. 1983 Jan;39(1):213–219. doi: 10.1128/iai.39.1.213-219.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  31. O'Brien R. M., Lucas P. C., Forest C. D., Magnuson M. A., Granner D. K. Identification of a sequence in the PEPCK gene that mediates a negative effect of insulin on transcription. Science. 1990 Aug 3;249(4968):533–537. doi: 10.1126/science.2166335. [DOI] [PubMed] [Google Scholar]
  32. Petersen D. D., Koch S. R., Granner D. K. 3' noncoding region of phosphoenolpyruvate carboxykinase mRNA contains a glucocorticoid-responsive mRNA-stabilizing element. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7800–7804. doi: 10.1073/pnas.86.20.7800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rippe D. F., Berry L. J. Study of inhibition of induction of phosphoenolpyruvate carboxykinase by endotoxin with radial immunodiffusion. Infect Immun. 1972 Nov;6(5):766–772. doi: 10.1128/iai.6.5.766-772.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sandler S., Bendtzen K., Eizirik D. L., Welsh M. Interleukin-6 affects insulin secretion and glucose metabolism of rat pancreatic islets in vitro. Endocrinology. 1990 Feb;126(2):1288–1294. doi: 10.1210/endo-126-2-1288. [DOI] [PubMed] [Google Scholar]
  35. Sasaki K., Cripe T. P., Koch S. R., Andreone T. L., Petersen D. D., Beale E. G., Granner D. K. Multihormonal regulation of phosphoenolpyruvate carboxykinase gene transcription. The dominant role of insulin. J Biol Chem. 1984 Dec 25;259(24):15242–15251. [PubMed] [Google Scholar]
  36. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  37. Smith S. M., Snyder I. S. Effect of lipopolysaccharide and lipid A on mouse liver pyruvate kinase activity. Infect Immun. 1975 Nov;12(5):993–998. doi: 10.1128/iai.12.5.993-998.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Stith R. D., McCallum R. E. Failure of glucagon to induce hepatic phosphoenolpyruvate carboxykinase in endotoxic shock. Circ Shock. 1988 Nov;26(3):267–272. [PubMed] [Google Scholar]
  39. Taga T., Kawanishi Y., Hardy R. R., Hirano T., Kishimoto T. Receptors for B cell stimulatory factor 2. Quantitation, specificity, distribution, and regulation of their expression. J Exp Med. 1987 Oct 1;166(4):967–981. doi: 10.1084/jem.166.4.967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Van Snick J., Cayphas S., Vink A., Uyttenhove C., Coulie P. G., Rubira M. R., Simpson R. J. Purification and NH2-terminal amino acid sequence of a T-cell-derived lymphokine with growth factor activity for B-cell hybridomas. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9679–9683. doi: 10.1073/pnas.83.24.9679. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES