Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Nov;15(11):6003–6012. doi: 10.1128/mcb.15.11.6003

Involvement of an AP-1 complex in zone-specific expression of the CYP11B1 gene in the rat adrenal cortex.

K Mukai 1, F Mitani 1, H Shimada 1, Y Ishimura 1
PMCID: PMC230852  PMID: 7565753

Abstract

The CYP11B1 gene, which encodes steroid 11 beta-monooxygenase, which is responsible for the synthesis of cortisol and corticosterone, the major glucocorticoids in mammals, is expressed specifically in the zona fasciculata of the adrenal cortex. We have analyzed the promoter region of the rat CYP11B1 gene by using a transient-expression system with adrenocortical Y1 cells and have identified a positive regulatory region. The region contained two adjacent sites for the binding of Y1-cell nuclear proteins: the binding site for an AP-1 transcription factor composed of JunD and a Fos-related protein, and the site for Ad4-binding protein (Ad4BP). The binding of the AP-1 factor to the regulatory region had a suppressive effect on that of Ad4BP in the nuclear extracts. Mutational analyses revealed that the transcriptional activation of the CYP11B1 gene promoter in Y1 cells was attributable to the AP-1 site but not to the Ad4 site. Subsequently, nuclear extracts of the zona fasciculata cells from the rat adrenal cortex were found to contain both AP-1 factor and Ad4BP, whose binding properties to the regulatory region were almost identical to those of the two factors in the Y1-cell nuclear extracts. Moreover, immunohistochemical analyses of rat adrenal cortices showed that the AP-1 factor was present in the nuclei of CYP11B1-expressing cells in the zona fasciculata but not in the nuclei of cells in the other zones. From these results, we propose that the AP-1 transcription factor found in this study plays an important role in the zone-specific expression of the CYP11B1 gene in rat adrenal cortex.

Full Text

The Full Text of this article is available as a PDF (926.1 KB).

Selected References

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

  1. Angel P., Karin M. The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta. 1991 Dec 10;1072(2-3):129–157. doi: 10.1016/0304-419x(91)90011-9. [DOI] [PubMed] [Google Scholar]
  2. Bogerd A. M., Franklin A., Rice D. A., Schimmer B. P., Parker K. L. Identification and characterization of two upstream elements that regulate adrenocortical expression of steroid 11 beta-hydroxylase. Mol Endocrinol. 1990 Jun;4(6):845–850. doi: 10.1210/mend-4-6-845. [DOI] [PubMed] [Google Scholar]
  3. Bohmann D., Bos T. J., Admon A., Nishimura T., Vogt P. K., Tjian R. Human proto-oncogene c-jun encodes a DNA binding protein with structural and functional properties of transcription factor AP-1. Science. 1987 Dec 4;238(4832):1386–1392. doi: 10.1126/science.2825349. [DOI] [PubMed] [Google Scholar]
  4. Boise L. H., Petryniak B., Mao X., June C. H., Wang C. Y., Lindsten T., Bravo R., Kovary K., Leiden J. M., Thompson C. B. The NFAT-1 DNA binding complex in activated T cells contains Fra-1 and JunB. Mol Cell Biol. 1993 Mar;13(3):1911–1919. doi: 10.1128/mcb.13.3.1911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boutillier A. L., Sassone-Corsi P., Loeffler J. P. The protooncogene c-fos is induced by corticotropin-releasing factor and stimulates proopiomelanocortin gene transcription in pituitary cells. Mol Endocrinol. 1991 Sep;5(9):1301–1310. doi: 10.1210/mend-5-9-1301. [DOI] [PubMed] [Google Scholar]
  6. Carrasco D., Bravo R. Tissue-specific expression of the fos-related transcription factor fra-2 during mouse development. Oncogene. 1995 Mar 16;10(6):1069–1079. [PubMed] [Google Scholar]
  7. Castellazzi M., Spyrou G., La Vista N., Dangy J. P., Piu F., Yaniv M., Brun G. Overexpression of c-jun, junB, or junD affects cell growth differently. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8890–8894. doi: 10.1073/pnas.88.20.8890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen D. R., Ferreira P. C., Gentz R., Franza B. R., Jr, Curran T. The product of a fos-related gene, fra-1, binds cooperatively to the AP-1 site with Jun: transcription factor AP-1 is comprised of multiple protein complexes. Genes Dev. 1989 Feb;3(2):173–184. doi: 10.1101/gad.3.2.173. [DOI] [PubMed] [Google Scholar]
  9. Curnow K. M., Tusie-Luna M. T., Pascoe L., Natarajan R., Gu J. L., Nadler J. L., White P. C. The product of the CYP11B2 gene is required for aldosterone biosynthesis in the human adrenal cortex. Mol Endocrinol. 1991 Oct;5(10):1513–1522. doi: 10.1210/mend-5-10-1513. [DOI] [PubMed] [Google Scholar]
  10. Curran T., Van Beveren C., Verma I. M. Viral and cellular fos proteins are complexed with a 39,000-dalton cellular protein. Mol Cell Biol. 1985 Jan;5(1):167–172. doi: 10.1128/mcb.5.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Togni P., Niman H., Raymond V., Sawchenko P., Verma I. M. Detection of fos protein during osteogenesis by monoclonal antibodies. Mol Cell Biol. 1988 May;8(5):2251–2256. doi: 10.1128/mcb.8.5.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Domalik L. J., Chaplin D. D., Kirkman M. S., Wu R. C., Liu W. W., Howard T. A., Seldin M. F., Parker K. L. Different isozymes of mouse 11 beta-hydroxylase produce mineralocorticoids and glucocorticoids. Mol Endocrinol. 1991 Dec;5(12):1853–1861. doi: 10.1210/mend-5-12-1853. [DOI] [PubMed] [Google Scholar]
  13. Evans R. M. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. doi: 10.1126/science.3283939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hilberg F., Aguzzi A., Howells N., Wagner E. F. c-jun is essential for normal mouse development and hepatogenesis. Nature. 1993 Sep 9;365(6442):179–181. doi: 10.1038/365179a0. [DOI] [PubMed] [Google Scholar]
  16. Honda S., Morohashi K., Nomura M., Takeya H., Kitajima M., Omura T. Ad4BP regulating steroidogenic P-450 gene is a member of steroid hormone receptor superfamily. J Biol Chem. 1993 Apr 5;268(10):7494–7502. [PubMed] [Google Scholar]
  17. Ikeda Y., Lala D. S., Luo X., Kim E., Moisan M. P., Parker K. L. Characterization of the mouse FTZ-F1 gene, which encodes a key regulator of steroid hydroxylase gene expression. Mol Endocrinol. 1993 Jul;7(7):852–860. doi: 10.1210/mend.7.7.8413309. [DOI] [PubMed] [Google Scholar]
  18. Ikeda Y., Shen W. H., Ingraham H. A., Parker K. L. Developmental expression of mouse steroidogenic factor-1, an essential regulator of the steroid hydroxylases. Mol Endocrinol. 1994 May;8(5):654–662. doi: 10.1210/mend.8.5.8058073. [DOI] [PubMed] [Google Scholar]
  19. Johnson R. S., Spiegelman B. M., Papaioannou V. Pleiotropic effects of a null mutation in the c-fos proto-oncogene. Cell. 1992 Nov 13;71(4):577–586. doi: 10.1016/0092-8674(92)90592-z. [DOI] [PubMed] [Google Scholar]
  20. Johnson R. S., van Lingen B., Papaioannou V. E., Spiegelman B. M. A null mutation at the c-jun locus causes embryonic lethality and retarded cell growth in culture. Genes Dev. 1993 Jul;7(7B):1309–1317. doi: 10.1101/gad.7.7b.1309. [DOI] [PubMed] [Google Scholar]
  21. Kagawa N., Waterman M. R. Evidence that an adrenal-specific nuclear protein regulates the cAMP responsiveness of the human CYP21B (P450C21) gene. J Biol Chem. 1991 Jun 15;266(17):11199–11204. [PubMed] [Google Scholar]
  22. Kagawa N., Waterman M. R. Purification and characterization of a transcription factor which appears to regulate cAMP responsiveness of the human CYP21B gene. J Biol Chem. 1992 Dec 15;267(35):25213–25219. [PubMed] [Google Scholar]
  23. Kawamoto T., Mitsuuchi Y., Toda K., Yokoyama Y., Miyahara K., Miura S., Ohnishi T., Ichikawa Y., Nakao K., Imura H. Role of steroid 11 beta-hydroxylase and steroid 18-hydroxylase in the biosynthesis of glucocorticoids and mineralocorticoids in humans. Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1458–1462. doi: 10.1073/pnas.89.4.1458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kirita S., Hashimoto T., Kitajima M., Honda S., Morohashi K., Omura T. Structural analysis of multiple bovine P-450(11 beta) genes and their promoter activities. J Biochem. 1990 Dec;108(6):1030–1041. doi: 10.1093/oxfordjournals.jbchem.a123302. [DOI] [PubMed] [Google Scholar]
  25. Lala D. S., Rice D. A., Parker K. L. Steroidogenic factor I, a key regulator of steroidogenic enzyme expression, is the mouse homolog of fushi tarazu-factor I. Mol Endocrinol. 1992 Aug;6(8):1249–1258. doi: 10.1210/mend.6.8.1406703. [DOI] [PubMed] [Google Scholar]
  26. Lauber M., Muller J. Purification and characterization of two distinct forms of rat adrenal cytochrome P450(11) beta: functional and structural aspects. Arch Biochem Biophys. 1989 Oct;274(1):109–119. doi: 10.1016/0003-9861(89)90421-9. [DOI] [PubMed] [Google Scholar]
  27. Luo X., Ikeda Y., Parker K. L. A cell-specific nuclear receptor is essential for adrenal and gonadal development and sexual differentiation. Cell. 1994 May 20;77(4):481–490. doi: 10.1016/0092-8674(94)90211-9. [DOI] [PubMed] [Google Scholar]
  28. Lynch J. P., Lala D. S., Peluso J. J., Luo W., Parker K. L., White B. A. Steroidogenic factor 1, an orphan nuclear receptor, regulates the expression of the rat aromatase gene in gonadal tissues. Mol Endocrinol. 1993 Jun;7(6):776–786. doi: 10.1210/mend.7.6.8395654. [DOI] [PubMed] [Google Scholar]
  29. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  30. Mitani F., Suzuki H., Hata J., Ogishima T., Shimada H., Ishimura Y. A novel cell layer without corticosteroid-synthesizing enzymes in rat adrenal cortex: histochemical detection and possible physiological role. Endocrinology. 1994 Jul;135(1):431–438. doi: 10.1210/endo.135.1.8013381. [DOI] [PubMed] [Google Scholar]
  31. Moore C. C., Brentano S. T., Miller W. L. Human P450scc gene transcription is induced by cyclic AMP and repressed by 12-O-tetradecanoylphorbol-13-acetate and A23187 through independent cis elements. Mol Cell Biol. 1990 Nov;10(11):6013–6023. doi: 10.1128/mcb.10.11.6013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Morohashi K., Honda S., Inomata Y., Handa H., Omura T. A common trans-acting factor, Ad4-binding protein, to the promoters of steroidogenic P-450s. J Biol Chem. 1992 Sep 5;267(25):17913–17919. [PubMed] [Google Scholar]
  33. Morohashi K., Iida H., Nomura M., Hatano O., Honda S., Tsukiyama T., Niwa O., Hara T., Takakusu A., Shibata Y. Functional difference between Ad4BP and ELP, and their distributions in steroidogenic tissues. Mol Endocrinol. 1994 May;8(5):643–653. doi: 10.1210/mend.8.5.8058072. [DOI] [PubMed] [Google Scholar]
  34. Morohashi K., Nonaka Y., Kirita S., Hatano O., Takakusu A., Okamoto M., Omura T. Enzymatic activities of P-450(11 beta)s expressed by two cDNAs in COS-7 cells. J Biochem. 1990 Apr;107(4):635–640. doi: 10.1093/oxfordjournals.jbchem.a123099. [DOI] [PubMed] [Google Scholar]
  35. Morohashi K., Zanger U. M., Honda S., Hara M., Waterman M. R., Omura T. Activation of CYP11A and CYP11B gene promoters by the steroidogenic cell-specific transcription factor, Ad4BP. Mol Endocrinol. 1993 Sep;7(9):1196–1204. doi: 10.1210/mend.7.9.8247022. [DOI] [PubMed] [Google Scholar]
  36. Mouw A. R., Rice D. A., Meade J. C., Chua S. C., White P. C., Schimmer B. P., Parker K. L. Structural and functional analysis of the promoter region of the gene encoding mouse steroid 11 beta-hydroxylase. J Biol Chem. 1989 Jan 15;264(2):1305–1309. [PubMed] [Google Scholar]
  37. Mukai K., Imai M., Shimada H., Ishimura Y. Isolation and characterization of rat CYP11B genes involved in late steps of mineralo- and glucocorticoid syntheses. J Biol Chem. 1993 Apr 25;268(12):9130–9137. [PubMed] [Google Scholar]
  38. Mukai K., Imai M., Shimada H., Okada Y., Ogishima T., Ishimura Y. Structural differences in 5'-flanking regions of rat cytochrome P-450aldo and P-450(11) beta genes. Biochem Biophys Res Commun. 1991 Nov 14;180(3):1187–1193. doi: 10.1016/s0006-291x(05)81321-x. [DOI] [PubMed] [Google Scholar]
  39. Nomura M., Morohashi K., Kirita S., Nonaka Y., Okamoto M., Nawata H., Omura T. Three forms of rat CYP11B genes: 11 beta-hydroxylase gene, aldosterone synthase gene, and a novel gene. J Biochem. 1993 Feb;113(2):144–152. doi: 10.1093/oxfordjournals.jbchem.a124018. [DOI] [PubMed] [Google Scholar]
  40. Ogishima T., Mitani F., Ishimura Y. Isolation of aldosterone synthase cytochrome P-450 from zona glomerulosa mitochondria of rat adrenal cortex. J Biol Chem. 1989 Jul 5;264(19):10935–10938. [PubMed] [Google Scholar]
  41. Ogishima T., Mitani F., Ishimura Y. Isolation of two distinct cytochromes P-45011 beta with aldosterone synthase activity from bovine adrenocortical mitochondria. J Biochem. 1989 Apr;105(4):497–499. doi: 10.1093/oxfordjournals.jbchem.a122694. [DOI] [PubMed] [Google Scholar]
  42. Ogishima T., Shibata H., Shimada H., Mitani F., Suzuki H., Saruta T., Ishimura Y. Aldosterone synthase cytochrome P-450 expressed in the adrenals of patients with primary aldosteronism. J Biol Chem. 1991 Jun 15;266(17):10731–10734. [PubMed] [Google Scholar]
  43. Ogishima T., Suzuki H., Hata J., Mitani F., Ishimura Y. Zone-specific expression of aldosterone synthase cytochrome P-450 and cytochrome P-45011 beta in rat adrenal cortex: histochemical basis for the functional zonation. Endocrinology. 1992 May;130(5):2971–2977. doi: 10.1210/endo.130.5.1572304. [DOI] [PubMed] [Google Scholar]
  44. Rice D. A., Mouw A. R., Bogerd A. M., Parker K. L. A shared promoter element regulates the expression of three steroidogenic enzymes. Mol Endocrinol. 1991 Oct;5(10):1552–1561. doi: 10.1210/mend-5-10-1552. [DOI] [PubMed] [Google Scholar]
  45. Rosenthal N. Identification of regulatory elements of cloned genes with functional assays. Methods Enzymol. 1987;152:704–720. doi: 10.1016/0076-6879(87)52075-4. [DOI] [PubMed] [Google Scholar]
  46. Ryseck R. P., Bravo R. c-JUN, JUN B, and JUN D differ in their binding affinities to AP-1 and CRE consensus sequences: effect of FOS proteins. Oncogene. 1991 Apr;6(4):533–542. [PubMed] [Google Scholar]
  47. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schimmer B. P. Adrenocortical Y1 cells. Methods Enzymol. 1979;58:570–574. doi: 10.1016/s0076-6879(79)58173-7. [DOI] [PubMed] [Google Scholar]
  49. 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]
  50. Simpson E. R., Waterman M. R. Regulation of the synthesis of steroidogenic enzymes in adrenal cortical cells by ACTH. Annu Rev Physiol. 1988;50:427–440. doi: 10.1146/annurev.ph.50.030188.002235. [DOI] [PubMed] [Google Scholar]
  51. Sonnenberg J. L., Macgregor-Leon P. F., Curran T., Morgan J. I. Dynamic alterations occur in the levels and composition of transcription factor AP-1 complexes after seizure. Neuron. 1989 Sep;3(3):359–365. doi: 10.1016/0896-6273(89)90260-2. [DOI] [PubMed] [Google Scholar]
  52. Suzuki T., Murakami M., Onai N., Fukuda E., Hashimoto Y., Sonobe M. H., Kameda T., Ichinose M., Miki K., Iba H. Analysis of AP-1 function in cellular transformation pathways. J Virol. 1994 Jun;68(6):3527–3535. doi: 10.1128/jvi.68.6.3527-3535.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Takayama K., Morohashi K., Honda S., Hara N., Omura T. Contribution of Ad4BP, a steroidogenic cell-specific transcription factor, to regulation of the human CYP11A and bovine CYP11B genes through their distal promoters. J Biochem. 1994 Jul;116(1):193–203. doi: 10.1093/oxfordjournals.jbchem.a124493. [DOI] [PubMed] [Google Scholar]
  54. Wang Z. Q., Ovitt C., Grigoriadis A. E., Möhle-Steinlein U., Rüther U., Wagner E. F. Bone and haematopoietic defects in mice lacking c-fos. Nature. 1992 Dec 24;360(6406):741–745. doi: 10.1038/360741a0. [DOI] [PubMed] [Google Scholar]
  55. Zerial M., Toschi L., Ryseck R. P., Schuermann M., Müller R., Bravo R. The product of a novel growth factor activated gene, fos B, interacts with JUN proteins enhancing their DNA binding activity. EMBO J. 1989 Mar;8(3):805–813. doi: 10.1002/j.1460-2075.1989.tb03441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES