Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Feb 25;20(4):825–829. doi: 10.1093/nar/20.4.825

Glucocorticoid induction of CRE-binding protein isoform mRNAs in rat C6 glioma cells.

R A Jungmann 1, X S Wang 1, D M Milkowski 1, M L Short 1
PMCID: PMC312024  PMID: 1531873

Abstract

Mammalian cells express several distinct isoforms of transcription factor CREB (cAMP-responsive element binding protein). At least two forms, alpha- and delta CREB, arise through alternative splicing of the CREB gene transcript. In this communication we demonstrate that the mRNAs of several CREB isoforms are expressed in rat C6 glioma cells and that the intracellular levels of these mRNAs are markedly induced by the synthetic glucocorticoid dexamethasone. Nuclear run-off assays show that the induction occurs, at least in part, through a transcriptional mechanism. The enhanced cellular levels of CREB mRNAs are accompanied by increased CREB protein and CRE-binding activity of nuclear extracts as evaluated by immunoblot and Southwestern blot assays.

Full text

PDF
825

Images in this article

Selected References

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

  1. Akerblom I. E., Slater E. P., Beato M., Baxter J. D., Mellon P. L. Negative regulation by glucocorticoids through interference with a cAMP responsive enhancer. Science. 1988 Jul 15;241(4863):350–353. doi: 10.1126/science.2838908. [DOI] [PubMed] [Google Scholar]
  2. Beato M. Transcriptional control by nuclear receptors. FASEB J. 1991 Apr;5(7):2044–2051. doi: 10.1096/fasebj.5.7.2010057. [DOI] [PubMed] [Google Scholar]
  3. Berkowitz L. A., Gilman M. Z. Two distinct forms of active transcription factor CREB (cAMP response element binding protein). Proc Natl Acad Sci U S A. 1990 Jul;87(14):5258–5262. doi: 10.1073/pnas.87.14.5258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DeVellis J., Brooker G. Reversal of catecholamine refractoriness by inhibitors of RNA and protein synthesis. Science. 1974 Dec 27;186(4170):1221–1223. doi: 10.1126/science.186.4170.1221. [DOI] [PubMed] [Google Scholar]
  5. Derda D. F., Miles M. F., Schweppe J. S., Jungmann R. A. Cyclic AMP regulation of lactate dehydrogenase. Isoproterenol and N6,O2'-dibutyryl cyclic AMP increase the levels of lactate dehydrogenase-5 isozyme and its messenger RNA in rat C6 glioma cells. J Biol Chem. 1980 Dec 10;255(23):11112–11121. [PubMed] [Google Scholar]
  6. Diamond M. I., Miner J. N., Yoshinaga S. K., Yamamoto K. R. Transcription factor interactions: selectors of positive or negative regulation from a single DNA element. Science. 1990 Sep 14;249(4974):1266–1272. doi: 10.1126/science.2119054. [DOI] [PubMed] [Google Scholar]
  7. Foulkes N. S., Borrelli E., Sassone-Corsi P. CREM gene: use of alternative DNA-binding domains generates multiple antagonists of cAMP-induced transcription. Cell. 1991 Feb 22;64(4):739–749. doi: 10.1016/0092-8674(91)90503-q. [DOI] [PubMed] [Google Scholar]
  8. Fukasawa K. M., Li S. S. Nucleotide sequence of the putative regulatory region of mouse lactate dehydrogenase-A gene. Biochem J. 1986 Apr 15;235(2):435–439. doi: 10.1042/bj2350435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gonzalez G. A., Yamamoto K. K., Fischer W. H., Karr D., Menzel P., Biggs W., 3rd, Vale W. W., Montminy M. R. A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence. Nature. 1989 Feb 23;337(6209):749–752. doi: 10.1038/337749a0. [DOI] [PubMed] [Google Scholar]
  10. Habener J. F. Cyclic AMP response element binding proteins: a cornucopia of transcription factors. Mol Endocrinol. 1990 Aug;4(8):1087–1094. doi: 10.1210/mend-4-8-1087. [DOI] [PubMed] [Google Scholar]
  11. Harrison J. J., Schwoch G., Schweppe J. S., Jungmann R. A. Phosphorylative modification of histone H1 subspecies following isoproterenol and N6,O2'-dibutyryl cyclic AMP stimulation of rat C6 glioma cells. J Biol Chem. 1982 Nov 25;257(22):13602–13609. [PubMed] [Google Scholar]
  12. Hoeffler J. P., Meyer T. E., Waeber G., Habener J. F. Multiple adenosine 3',5'-cyclic [corrected] monophosphate response element DNA-binding proteins generated by gene diversification and alternative exon splicing. Mol Endocrinol. 1990 Jun;4(6):920–930. doi: 10.1210/mend-4-6-920. [DOI] [PubMed] [Google Scholar]
  13. Jungmann R. A., Kelley D. C., Miles M. F., Milkowski D. M. Cyclic AMP regulation of lactate dehydrogenase. Isoproterenol and N6,O2-dibutyryl cyclic amp increase the rate of transcription and change the stability of lactate dehydrogenase a subunit messenger RNA in rat C6 glioma cells. J Biol Chem. 1983 Apr 25;258(8):5312–5318. [PubMed] [Google Scholar]
  14. Kuettel M. R., Squinto S. P., Kwast-Welfeld J., Schwoch G., Schweppe J. S., Jungmann R. A. Localization of nuclear subunits of cyclic AMP-dependent protein kinase by the immunocolloidal gold method. J Cell Biol. 1985 Sep;101(3):965–975. doi: 10.1083/jcb.101.3.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kwast-Welfeld J., Soong C. J., Short M. L., Jungmann R. A. Identification of rat ovarian nuclear factors that interact with the cAMP-inducible lactate dehydrogenase A subunit promoter. J Biol Chem. 1989 Apr 25;264(12):6941–6947. [PubMed] [Google Scholar]
  16. Liu A. Y., Greengard P. Regulation by steroid hormones of phosphorylation of specific protein common to several target organs. Proc Natl Acad Sci U S A. 1976 Feb;73(2):568–572. doi: 10.1073/pnas.73.2.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGinnis J. F., de Vellis J. Glucocorticoid regulation in rat brain cell cultures. Hydrocortisone increases the rate of synthesis of glycerol phosphate dehydrogenase in C6 glioma cells. J Biol Chem. 1978 Dec 10;253(23):8493–8492. [PubMed] [Google Scholar]
  18. Miles M. F., Hung P., Jungmann R. A. Cyclic AMP regulation of lactate dehydrogenase. Quantitation of lactate dehydrogenase M-subunit messenger RNA in isoproterenol-and N6,O2'-dibutyryl cyclic AMP-stimulated rat C6 glioma cells by hybridization analysis using a cloned cDNA probe. J Biol Chem. 1981 Dec 10;256(23):12545–12552. [PubMed] [Google Scholar]
  19. Oro A. E., Hollenberg S. M., Evans R. M. Transcriptional inhibition by a glucocorticoid receptor-beta-galactosidase fusion protein. Cell. 1988 Dec 23;55(6):1109–1114. doi: 10.1016/0092-8674(88)90255-3. [DOI] [PubMed] [Google Scholar]
  20. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  21. Shapiro D. J., Sharp P. A., Wahli W. W., Keller M. J. A high-efficiency HeLa cell nuclear transcription extract. DNA. 1988 Jan-Feb;7(1):47–55. doi: 10.1089/dna.1988.7.47. [DOI] [PubMed] [Google Scholar]
  22. Short M. L., Manohar C. F., Furtado M. R., Ghadge G. D., Wolinsky S. M., Thimmapaya B., Jungmann R. A. Nucleotide and derived amino-acid sequences of the CRE-binding proteins from rat C6 glioma and HeLa cells. Nucleic Acids Res. 1991 Aug 11;19(15):4290–4290. doi: 10.1093/nar/19.15.4290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Short S., Short M. L., Milkowski D. M., Jungmann R. A. Functional analysis of cis- and trans-regulatory elements of the lactate dehydrogenase A subunit promoter by in vitro transcription. J Biol Chem. 1991 Nov 25;266(33):22164–22172. [PubMed] [Google Scholar]
  24. Waeber G., Meyer T. E., LeSieur M., Hermann H. L., Gérard N., Habener J. F. Developmental stage-specific expression of cyclic adenosine 3',5'-monophosphate response element-binding protein CREB during spermatogenesis involves alternative exon splicing. Mol Endocrinol. 1991 Oct;5(10):1418–1430. doi: 10.1210/mend-5-10-1418. [DOI] [PubMed] [Google Scholar]
  25. Yamamoto K. K., Gonzalez G. A., Biggs W. H., 3rd, Montminy M. R. Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature. 1988 Aug 11;334(6182):494–498. doi: 10.1038/334494a0. [DOI] [PubMed] [Google Scholar]
  26. Yamamoto K. K., Gonzalez G. A., Menzel P., Rivier J., Montminy M. R. Characterization of a bipartite activator domain in transcription factor CREB. Cell. 1990 Feb 23;60(4):611–617. doi: 10.1016/0092-8674(90)90664-z. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES