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. 1996 Feb;16(2):584–592. doi: 10.1128/mcb.16.2.584

A composite element binding the vitamin D receptor, retinoid X receptor alpha, and a member of the CTF/NF-1 family of transcription factors mediates the vitamin D responsiveness of the c-fos promoter.

G A Candeliere 1, P W Jurutka 1, M R Haussler 1, R St-Arnaud 1
PMCID: PMC231037  PMID: 8552086

Abstract

The hormonal form of vitamin D, 1 alpha,25-dihydroxyvitamin D3 [1,25- (OH)2D3], transiently stimulates the transcription of the c-fos proto-oncogene in osteoblastic cells. We have identified and characterized a vitamin D response element (VDRE) in the promoter of c-fos. The 1,25-(OH)2D3-responsive region was delineated between residues -178 and -144 upstream of the c-fos transcription start site. A mutation that inhibited binding to the sequence concomitantly abolished 1,25-(OH)2D3-induced transcriptional responsiveness; similarly, cloning to the site upstream of a heterologous promoter conferred copy-number-dependent vitamin D responsiveness to a reporter gene, demonstrating that we have identified a functional response element. The structure of the c-fos VDRE was found to be unusual. Mutational analysis revealed that the c-fos VDRE does not conform to the direct repeat configuration in which hexameric core-binding sites are spaced by a few nucleotide residues. In contrast, the entire 36-bp sequence was essential for binding. We identified the vitamin D receptor and the retinoid X receptor alpha as components of the complex that bound the c-fos VDRE. However, our results also show that a putative CCAAT-binding transcription factor/nuclear factor 1 (CTF/NF-1) family member bound the response element in conjunction with the nuclear hormone receptors. The expression of this CTF/NF-1 family member appeared restricted to bone cells. These data hint at new molecular mechanisms of action for vitamin D.

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Selected References

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  1. Abate C., Curran T. Encounters with Fos and Jun on the road to AP-1. Semin Cancer Biol. 1990 Feb;1(1):19–26. [PubMed] [Google Scholar]
  2. Breen E. C., van Wijnen A. J., Lian J. B., Stein G. S., Stein J. L. In vivo occupancy of the vitamin D responsive element in the osteocalcin gene supports vitamin D-dependent transcriptional upregulation in intact cells. Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12902–12906. doi: 10.1073/pnas.91.26.12902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Candeliere G. A., Prud'homme J., St-Arnaud R. Differential stimulation of fos and jun family members by calcitriol in osteoblastic cells. Mol Endocrinol. 1991 Dec;5(12):1780–1788. doi: 10.1210/mend-5-12-1780. [DOI] [PubMed] [Google Scholar]
  4. Cao X., Ross F. P., Zhang L., MacDonald P. N., Chappel J., Teitelbaum S. L. Cloning of the promoter for the avian integrin beta 3 subunit gene and its regulation by 1,25-dihydroxyvitamin D3. J Biol Chem. 1993 Dec 25;268(36):27371–27380. [PubMed] [Google Scholar]
  5. Carlberg C., Bendik I., Wyss A., Meier E., Sturzenbecker L. J., Grippo J. F., Hunziker W. Two nuclear signalling pathways for vitamin D. Nature. 1993 Feb 18;361(6413):657–660. doi: 10.1038/361657a0. [DOI] [PubMed] [Google Scholar]
  6. Corthésy B., Cardinaux J. R., Claret F. X., Wahli W. A nuclear factor I-like activity and a liver-specific repressor govern estrogen-regulated in vitro transcription from the Xenopus laevis vitellogenin B1 promoter. Mol Cell Biol. 1989 Dec;9(12):5548–5562. doi: 10.1128/mcb.9.12.5548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Darwish H. M., DeLuca H. F. Identification of a 1,25-dihydroxyvitamin D3-response element in the 5'-flanking region of the rat calbindin D-9k gene. Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):603–607. doi: 10.1073/pnas.89.2.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Demay M. B., Gerardi J. M., DeLuca H. F., Kronenberg H. M. DNA sequences in the rat osteocalcin gene that bind the 1,25-dihydroxyvitamin D3 receptor and confer responsiveness to 1,25-dihydroxyvitamin D3. Proc Natl Acad Sci U S A. 1990 Jan;87(1):369–373. doi: 10.1073/pnas.87.1.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dony C., Gruss P. Proto-oncogene c-fos expression in growth regions of fetal bone and mesodermal web tissue. Nature. 1987 Aug 20;328(6132):711–714. doi: 10.1038/328711a0. [DOI] [PubMed] [Google Scholar]
  12. Fuller P. J. The steroid receptor superfamily: mechanisms of diversity. FASEB J. 1991 Dec;5(15):3092–3099. doi: 10.1096/fasebj.5.15.1743440. [DOI] [PubMed] [Google Scholar]
  13. Gil G., Smith J. R., Goldstein J. L., Slaughter C. A., Orth K., Brown M. S., Osborne T. F. Multiple genes encode nuclear factor 1-like proteins that bind to the promoter for 3-hydroxy-3-methylglutaryl-coenzyme A reductase. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8963–8967. doi: 10.1073/pnas.85.23.8963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gill R. K., Christakos S. Identification of sequence elements in mouse calbindin-D28k gene that confer 1,25-dihydroxyvitamin D3- and butyrate-inducible responses. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2984–2988. doi: 10.1073/pnas.90.7.2984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Glass C. K., Holloway J. M., Devary O. V., Rosenfeld M. G. The thyroid hormone receptor binds with opposite transcriptional effects to a common sequence motif in thyroid hormone and estrogen response elements. Cell. 1988 Jul 29;54(3):313–323. doi: 10.1016/0092-8674(88)90194-8. [DOI] [PubMed] [Google Scholar]
  16. Hanazawa S., Takeshita A., Amano S., Semba T., Nirazuka T., Katoh H., Kitano S. Tumor necrosis factor-alpha induces expression of monocyte chemoattractant JE via fos and jun genes in clonal osteoblastic MC3T3-E1 cells. J Biol Chem. 1993 May 5;268(13):9526–9532. [PubMed] [Google Scholar]
  17. Heyman R. A., Mangelsdorf D. J., Dyck J. A., Stein R. B., Eichele G., Evans R. M., Thaller C. 9-cis retinoic acid is a high affinity ligand for the retinoid X receptor. Cell. 1992 Jan 24;68(2):397–406. doi: 10.1016/0092-8674(92)90479-v. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Jones K. A., Kadonaga J. T., Rosenfeld P. J., Kelly T. J., Tjian R. A cellular DNA-binding protein that activates eukaryotic transcription and DNA replication. Cell. 1987 Jan 16;48(1):79–89. doi: 10.1016/0092-8674(87)90358-8. [DOI] [PubMed] [Google Scholar]
  20. Jones K. A., Yamamoto K. R., Tjian R. Two distinct transcription factors bind to the HSV thymidine kinase promoter in vitro. Cell. 1985 Sep;42(2):559–572. doi: 10.1016/0092-8674(85)90113-8. [DOI] [PubMed] [Google Scholar]
  21. Kerner S. A., Scott R. A., Pike J. W. Sequence elements in the human osteocalcin gene confer basal activation and inducible response to hormonal vitamin D3. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4455–4459. doi: 10.1073/pnas.86.12.4455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kliewer S. A., Umesono K., Mangelsdorf D. J., Evans R. M. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature. 1992 Jan 30;355(6359):446–449. doi: 10.1038/355446a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee K., Deeds J. D., Chiba S., Un-No M., Bond A. T., Segre G. V. Parathyroid hormone induces sequential c-fos expression in bone cells in vivo: in situ localization of its receptor and c-fos messenger ribonucleic acids. Endocrinology. 1994 Jan;134(1):441–450. doi: 10.1210/endo.134.1.8275957. [DOI] [PubMed] [Google Scholar]
  24. Levin A. A., Sturzenbecker L. J., Kazmer S., Bosakowski T., Huselton C., Allenby G., Speck J., Kratzeisen C., Rosenberger M., Lovey A. 9-cis retinoic acid stereoisomer binds and activates the nuclear receptor RXR alpha. Nature. 1992 Jan 23;355(6358):359–361. doi: 10.1038/355359a0. [DOI] [PubMed] [Google Scholar]
  25. Liao J., Ozono K., Sone T., McDonnell D. P., Pike J. W. Vitamin D receptor interaction with specific DNA requires a nuclear protein and 1,25-dihydroxyvitamin D3. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9751–9755. doi: 10.1073/pnas.87.24.9751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Liu M., Freedman L. P. Transcriptional synergism between the vitamin D3 receptor and other nonreceptor transcription factors. Mol Endocrinol. 1994 Dec;8(12):1593–1604. doi: 10.1210/mend.8.12.7708050. [DOI] [PubMed] [Google Scholar]
  27. MacDonald P. N., Dowd D. R., Nakajima S., Galligan M. A., Reeder M. C., Haussler C. A., Ozato K., Haussler M. R. Retinoid X receptors stimulate and 9-cis retinoic acid inhibits 1,25-dihydroxyvitamin D3-activated expression of the rat osteocalcin gene. Mol Cell Biol. 1993 Sep;13(9):5907–5917. doi: 10.1128/mcb.13.9.5907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mangelsdorf D. J., Borgmeyer U., Heyman R. A., Zhou J. Y., Ong E. S., Oro A. E., Kakizuka A., Evans R. M. Characterization of three RXR genes that mediate the action of 9-cis retinoic acid. Genes Dev. 1992 Mar;6(3):329–344. doi: 10.1101/gad.6.3.329. [DOI] [PubMed] [Google Scholar]
  29. Mermod N., O'Neill E. A., Kelly T. J., Tjian R. The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domain. Cell. 1989 Aug 25;58(4):741–753. doi: 10.1016/0092-8674(89)90108-6. [DOI] [PubMed] [Google Scholar]
  30. Merriman H. L., La Tour D., Linkhart T. A., Mohan S., Baylink D. J., Strong D. D. Insulin-like growth factor-I and insulin-like growth factor-II induce c-fos in mouse osteoblastic cells. Calcif Tissue Int. 1990 Apr;46(4):258–262. doi: 10.1007/BF02555005. [DOI] [PubMed] [Google Scholar]
  31. Miller A. D., Curran T., Verma I. M. c-fos protein can induce cellular transformation: a novel mechanism of activation of a cellular oncogene. Cell. 1984 Jan;36(1):51–60. doi: 10.1016/0092-8674(84)90073-4. [DOI] [PubMed] [Google Scholar]
  32. Morrison N. A., Shine J., Fragonas J. C., Verkest V., McMenemy M. L., Eisman J. A. 1,25-dihydroxyvitamin D-responsive element and glucocorticoid repression in the osteocalcin gene. Science. 1989 Dec 1;246(4934):1158–1161. doi: 10.1126/science.2588000. [DOI] [PubMed] [Google Scholar]
  33. Nishikawa J., Matsumoto M., Sakoda K., Kitaura M., Imagawa M., Nishihara T. Vitamin D receptor zinc finger region binds to a direct repeat as a dimer and discriminates the spacing number between each half-site. J Biol Chem. 1993 Sep 15;268(26):19739–19743. [PubMed] [Google Scholar]
  34. Noda M., Vogel R. L., Craig A. M., Prahl J., DeLuca H. F., Denhardt D. T. Identification of a DNA sequence responsible for binding of the 1,25-dihydroxyvitamin D3 receptor and 1,25-dihydroxyvitamin D3 enhancement of mouse secreted phosphoprotein 1 (SPP-1 or osteopontin) gene expression. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9995–9999. doi: 10.1073/pnas.87.24.9995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ozono K., Sone T., Pike J. W. The genomic mechanism of action of 1,25-dihydroxyvitamin D3. J Bone Miner Res. 1991 Oct;6(10):1021–1027. doi: 10.1002/jbmr.5650061002. [DOI] [PubMed] [Google Scholar]
  36. Rhodes S. J., Chen R., DiMattia G. E., Scully K. M., Kalla K. A., Lin S. C., Yu V. C., Rosenfeld M. G. A tissue-specific enhancer confers Pit-1-dependent morphogen inducibility and autoregulation on the pit-1 gene. Genes Dev. 1993 Jun;7(6):913–932. doi: 10.1101/gad.7.6.913. [DOI] [PubMed] [Google Scholar]
  37. Runkel L., Shaw P. E., Herrera R. E., Hipskind R. A., Nordheim A. Multiple basal promoter elements determine the level of human c-fos transcription. Mol Cell Biol. 1991 Mar;11(3):1270–1280. doi: 10.1128/mcb.11.3.1270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rüther U., Garber C., Komitowski D., Müller R., Wagner E. F. Deregulated c-fos expression interferes with normal bone development in transgenic mice. 1987 Jan 29-Feb 4Nature. 325(6103):412–416. doi: 10.1038/325412a0. [DOI] [PubMed] [Google Scholar]
  39. Santoro C., Mermod N., Andrews P. C., Tjian R. A family of human CCAAT-box-binding proteins active in transcription and DNA replication: cloning and expression of multiple cDNAs. Nature. 1988 Jul 21;334(6179):218–224. doi: 10.1038/334218a0. [DOI] [PubMed] [Google Scholar]
  40. Schräder M., Bendik I., Becker-André M., Carlberg C. Interaction between retinoic acid and vitamin D signaling pathways. J Biol Chem. 1993 Aug 25;268(24):17830–17836. [PubMed] [Google Scholar]
  41. Schräder M., Müller K. M., Carlberg C. Specificity and flexibility of vitamin D signaling. Modulation of the activation of natural vitamin D response elements by thyroid hormone. J Biol Chem. 1994 Feb 25;269(8):5501–5504. [PubMed] [Google Scholar]
  42. Schräder M., Müller K. M., Nayeri S., Kahlen J. P., Carlberg C. Vitamin D3-thyroid hormone receptor heterodimer polarity directs ligand sensitivity of transactivation. Nature. 1994 Aug 4;370(6488):382–386. doi: 10.1038/370382a0. [DOI] [PubMed] [Google Scholar]
  43. Slootweg M. C., van Genesen S. T., Otte A. P., Duursma S. A., Kruijer W. Activation of mouse osteoblast growth hormone receptor: c-fos oncogene expression independent of phosphoinositide breakdown and cyclic AMP. J Mol Endocrinol. 1990 Jun;4(3):265–274. doi: 10.1677/jme.0.0040265. [DOI] [PubMed] [Google Scholar]
  44. St-Arnaud R., Moir J. M. Wnt-1-inducing factor-1: a novel G/C box-binding transcription factor regulating the expression of Wnt-1 during neuroectodermal differentiation. Mol Cell Biol. 1993 Mar;13(3):1590–1598. doi: 10.1128/mcb.13.3.1590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tanese N., Pugh B. F., Tjian R. Coactivators for a proline-rich activator purified from the multisubunit human TFIID complex. Genes Dev. 1991 Dec;5(12A):2212–2224. doi: 10.1101/gad.5.12a.2212. [DOI] [PubMed] [Google Scholar]
  46. Toohey M. G., Lee J. W., Huang M., Peterson D. O. Functional elements of the steroid hormone-responsive promoter of mouse mammary tumor virus. J Virol. 1990 Sep;64(9):4477–4488. doi: 10.1128/jvi.64.9.4477-4488.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Umesono K., Murakami K. K., Thompson C. C., Evans R. M. Direct repeats as selective response elements for the thyroid hormone, retinoic acid, and vitamin D3 receptors. Cell. 1991 Jun 28;65(7):1255–1266. doi: 10.1016/0092-8674(91)90020-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Yu V. C., Delsert C., Andersen B., Holloway J. M., Devary O. V., När A. M., Kim S. Y., Boutin J. M., Glass C. K., Rosenfeld M. G. RXR beta: a coregulator that enhances binding of retinoic acid, thyroid hormone, and vitamin D receptors to their cognate response elements. Cell. 1991 Dec 20;67(6):1251–1266. doi: 10.1016/0092-8674(91)90301-e. [DOI] [PubMed] [Google Scholar]
  50. Zierold C., Darwish H. M., DeLuca H. F. Identification of a vitamin D-response element in the rat calcidiol (25-hydroxyvitamin D3) 24-hydroxylase gene. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):900–902. doi: 10.1073/pnas.91.3.900. [DOI] [PMC free article] [PubMed] [Google Scholar]

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