Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1995 Aug 25;23(16):3268–3274. doi: 10.1093/nar/23.16.3268

Calreticulin inhibits vitamin D3 signal transduction.

D G Wheeler 1, J Horsford 1, M Michalak 1, J H White 1, G N Hendy 1
PMCID: PMC307187  PMID: 7667104

Abstract

Calreticulin is a calcium binding protein present primarily in the lumen of the endoplasmic reticulum. However, it can also localize to the cytoplasm adjacent to the cell membrane where it binds integrins, and to the nucleus. Recent studies showed that calreticulin inhibits DNA binding and transcriptional activity of glucocorticoid, androgen and retinoic acid receptors. The DNA binding domains of nuclear receptors share a common motif based upon the amino acid sequence KVFFKR which has been implicated in the binding of calreticulin. The vitamin D receptor (VDR) DNA binding domain contains the related motif KgFFrR. Here we show that calreticulin blocks specific DNA binding by the isolated VDR DNA binding domain in DNA mobility shift assays. Importantly, calreticulin blocks specific DNA binding by the full length VDR-RXR heterodimers. By contrast, calreticulin had no effect on specific DNA binding by the transcription factor ATF-a delta which lacks a KVFFKR-like motif in its DNA binding domain. We further showed that overexpression of calreticulin in the rat osteoblast-like cell line (ROS 17/2.8) inhibited the 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] responsive transcriptional activation of a vitamin D-sensitive reporter gene, whereas the response to forskolin stimulation of a control promoter-reporter construct containing a cAMP response element (CRE), but no vitamin D response element (VDRE), was not affected by overexpression of calreticulin. Thus, calreticulin inhibits transcriptional activation by the VDR in vivo. Given the ubiquitous expression of calreticulin and the widespread expression of the VDR the studies described here may point to an important new mechanism whereby VDR mediated gene transcription can be modulated.

Full text

PDF
3268

Images in this article

3270Image
on p.3270
3271Image
on p.3271
3272Image
on p.3272

Selected References

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

  1. Baker A. R., McDonnell D. P., Hughes M., Crisp T. M., Mangelsdorf D. J., Haussler M. R., Pike J. W., Shine J., O'Malley B. W. Cloning and expression of full-length cDNA encoding human vitamin D receptor. Proc Natl Acad Sci U S A. 1988 May;85(10):3294–3298. doi: 10.1073/pnas.85.10.3294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Banerji J., Rusconi S., Schaffner W. Expression of a beta-globin gene is enhanced by remote SV40 DNA sequences. Cell. 1981 Dec;27(2 Pt 1):299–308. doi: 10.1016/0092-8674(81)90413-x. [DOI] [PubMed] [Google Scholar]
  3. Burns K., Atkinson E. A., Bleackley R. C., Michalak M. Calreticulin: from Ca2+ binding to control of gene expression. Trends Cell Biol. 1994 May;4(5):152–154. doi: 10.1016/0962-8924(94)90190-2. [DOI] [PubMed] [Google Scholar]
  4. Burns K., Duggan B., Atkinson E. A., Famulski K. S., Nemer M., Bleackley R. C., Michalak M. Modulation of gene expression by calreticulin binding to the glucocorticoid receptor. Nature. 1994 Feb 3;367(6462):476–480. doi: 10.1038/367476a0. [DOI] [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. Cheskis B., Freedman L. P. Ligand modulates the conversion of DNA-bound vitamin D3 receptor (VDR) homodimers into VDR-retinoid X receptor heterodimers. Mol Cell Biol. 1994 May;14(5):3329–3338. doi: 10.1128/mcb.14.5.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clemens T. L., Garrett K. P., Zhou X. Y., Pike J. W., Haussler M. R., Dempster D. W. Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. Endocrinology. 1988 Apr;122(4):1224–1230. doi: 10.1210/endo-122-4-1224. [DOI] [PubMed] [Google Scholar]
  8. Dedhar S., Rennie P. S., Shago M., Hagesteijn C. Y., Yang H., Filmus J., Hawley R. G., Bruchovsky N., Cheng H., Matusik R. J. Inhibition of nuclear hormone receptor activity by calreticulin. Nature. 1994 Feb 3;367(6462):480–483. doi: 10.1038/367480a0. [DOI] [PubMed] [Google Scholar]
  9. Demay M. B., Kiernan M. S., DeLuca H. F., Kronenberg H. M. Sequences in the human parathyroid hormone gene that bind the 1,25-dihydroxyvitamin D3 receptor and mediate transcriptional repression in response to 1,25-dihydroxyvitamin D3. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8097–8101. doi: 10.1073/pnas.89.17.8097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fawell S. E., Lees J. A., White R., Parker M. G. Characterization and colocalization of steroid binding and dimerization activities in the mouse estrogen receptor. Cell. 1990 Mar 23;60(6):953–962. doi: 10.1016/0092-8674(90)90343-d. [DOI] [PubMed] [Google Scholar]
  11. Ferrara J., McCuaig K., Hendy G. N., Uskokovic M., White J. H. Highly potent transcriptional activation by 16-ene derivatives of 1,25-dihydroxyvitamin D3. Lack of modulation by 9-cis-retinoic acid of response to 1,25-dihydroxyvitamin D3 or its derivatives. J Biol Chem. 1994 Jan 28;269(4):2971–2981. [PubMed] [Google Scholar]
  12. Gaire M., Chatton B., Kedinger C. Isolation and characterization of two novel, closely related ATF cDNA clones from HeLa cells. Nucleic Acids Res. 1990 Jun 25;18(12):3467–3473. doi: 10.1093/nar/18.12.3467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kumar V., Chambon P. The estrogen receptor binds tightly to its responsive element as a ligand-induced homodimer. Cell. 1988 Oct 7;55(1):145–156. doi: 10.1016/0092-8674(88)90017-7. [DOI] [PubMed] [Google Scholar]
  14. Kuno H., Kurian S. M., Hendy G. N., White J., deLuca H. F., Evans C. O., Nanes M. S. Inhibition of 1,25-dihydroxyvitamin D3 stimulated osteocalcin gene transcription by tumor necrosis factor-alpha: structural determinants within the vitamin D response element. Endocrinology. 1994 Jun;134(6):2524–2531. doi: 10.1210/endo.134.6.8194478. [DOI] [PubMed] [Google Scholar]
  15. Leventis R., Silvius J. R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles. Biochim Biophys Acta. 1990 Mar 30;1023(1):124–132. doi: 10.1016/0005-2736(90)90017-i. [DOI] [PubMed] [Google Scholar]
  16. MacDonald P. N., Dowd D. R., Haussler M. R. New insight into the structure and functions of the vitamin D receptor. Semin Nephrol. 1994 Mar;14(2):101–118. [PubMed] [Google Scholar]
  17. Mader S., Chambon P., White J. H. Defining a minimal estrogen receptor DNA binding domain. Nucleic Acids Res. 1993 Mar 11;21(5):1125–1132. doi: 10.1093/nar/21.5.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mader S., Leroy P., Chen J. Y., Chambon P. Multiple parameters control the selectivity of nuclear receptors for their response elements. Selectivity and promiscuity in response element recognition by retinoic acid receptors and retinoid X receptors. J Biol Chem. 1993 Jan 5;268(1):591–600. [PubMed] [Google Scholar]
  19. Michalak M., Milner R. E., Burns K., Opas M. Calreticulin. Biochem J. 1992 Aug 1;285(Pt 3):681–692. doi: 10.1042/bj2850681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milner R. E., Baksh S., Shemanko C., Carpenter M. R., Smillie L., Vance J. E., Opas M., Michalak M. Calreticulin, and not calsequestrin, is the major calcium binding protein of smooth muscle sarcoplasmic reticulum and liver endoplasmic reticulum. J Biol Chem. 1991 Apr 15;266(11):7155–7165. [PubMed] [Google Scholar]
  21. Montminy M. R., Sevarino K. A., Wagner J. A., Mandel G., Goodman R. H. Identification of a cyclic-AMP-responsive element within the rat somatostatin gene. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6682–6686. doi: 10.1073/pnas.83.18.6682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Opas M., Dziak E., Fliegel L., Michalak M. Regulation of expression and intracellular distribution of calreticulin, a major calcium binding protein of nonmuscle cells. J Cell Physiol. 1991 Oct;149(1):160–171. doi: 10.1002/jcp.1041490120. [DOI] [PubMed] [Google Scholar]
  23. Owen T. A., Bortell R., Yocum S. A., Smock S. L., Zhang M., Abate C., Shalhoub V., Aronin N., Wright K. L., van Wijnen A. J. Coordinate occupancy of AP-1 sites in the vitamin D-responsive and CCAAT box elements by Fos-Jun in the osteocalcin gene: model for phenotype suppression of transcription. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9990–9994. doi: 10.1073/pnas.87.24.9990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peleg S., Abruzzese R. V., Cooper C. W., Gagel R. F. Down-regulation of calcitonin gene transcription by vitamin D requires two widely separated enhancer sequences. Mol Endocrinol. 1993 Aug;7(8):999–1008. doi: 10.1210/mend.7.8.8232320. [DOI] [PubMed] [Google Scholar]
  25. Pike J. W. Vitamin D3 receptors: structure and function in transcription. Annu Rev Nutr. 1991;11:189–216. doi: 10.1146/annurev.nu.11.070191.001201. [DOI] [PubMed] [Google Scholar]
  26. Reichel H., Koeffler H. P., Norman A. W. The role of the vitamin D endocrine system in health and disease. N Engl J Med. 1989 Apr 13;320(15):980–991. doi: 10.1056/NEJM198904133201506. [DOI] [PubMed] [Google Scholar]
  27. Rojiani M. V., Finlay B. B., Gray V., Dedhar S. In vitro interaction of a polypeptide homologous to human Ro/SS-A antigen (calreticulin) with a highly conserved amino acid sequence in the cytoplasmic domain of integrin alpha subunits. Biochemistry. 1991 Oct 15;30(41):9859–9866. doi: 10.1021/bi00105a008. [DOI] [PubMed] [Google Scholar]
  28. Ross T. K., Darwish H. M., DeLuca H. F. Molecular biology of vitamin D action. Vitam Horm. 1994;49:281–326. doi: 10.1016/s0083-6729(08)61149-8. [DOI] [PubMed] [Google Scholar]
  29. Schüle R., Umesono K., Mangelsdorf D. J., Bolado J., Pike J. W., Evans R. M. Jun-Fos and receptors for vitamins A and D recognize a common response element in the human osteocalcin gene. Cell. 1990 May 4;61(3):497–504. doi: 10.1016/0092-8674(90)90531-i. [DOI] [PubMed] [Google Scholar]
  30. Sebag M., Henderson J., Rhim J., Kremer R. Relative resistance to 1,25-dihydroxyvitamin D3 in a keratinocyte model of tumor progression. J Biol Chem. 1992 Jun 15;267(17):12162–12167. [PubMed] [Google Scholar]
  31. Stein G. S., Lian J. B. Molecular mechanisms mediating proliferation/differentiation interrelationships during progressive development of the osteoblast phenotype. Endocr Rev. 1993 Aug;14(4):424–442. doi: 10.1210/edrv-14-4-424. [DOI] [PubMed] [Google Scholar]
  32. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  33. Stumpf W. E., Sar M., Reid F. A., Tanaka Y., DeLuca H. F. Target cells for 1,25-dihydroxyvitamin D3 in intestinal tract, stomach, kidney, skin, pituitary, and parathyroid. Science. 1979 Dec 7;206(4423):1188–1190. doi: 10.1126/science.505004. [DOI] [PubMed] [Google Scholar]
  34. Tora L., White J., Brou C., Tasset D., Webster N., Scheer E., Chambon P. The human estrogen receptor has two independent nonacidic transcriptional activation functions. Cell. 1989 Nov 3;59(3):477–487. doi: 10.1016/0092-8674(89)90031-7. [DOI] [PubMed] [Google Scholar]
  35. 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]

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

RESOURCES