Abstract
The human osteocalcin gene is regulated in mammalian osteoblasts by 1,25(OH)2D3-dependent and -independent mechanisms. The sequences responsible for this activity have been mapped to within the -1339 region of the gene. We show here that this enhancer region functions analogously in Saccharomyces cerevisiae cells engineered to produce active 1,25(OH)2D3 receptor. When fused to the proximal promoter elements of the yeast iso-1-cytochrome c gene, the enhancer demonstrated substantial promoter activity. This activity was elevated further by 1,25(OH)2D3 when the reporter constructs were assayed in cells containing the 1,25(OH)2D3 receptor. This system affords a model for 1,25(OH)2D3 action and represents a simple assay system that will enable definition of the important cis-acting regulatory sequences within the osteocalcin gene and identification of their cognate transcription factors.
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- Bachmair A., Finley D., Varshavsky A. In vivo half-life of a protein is a function of its amino-terminal residue. Science. 1986 Oct 10;234(4773):179–186. doi: 10.1126/science.3018930. [DOI] [PubMed] [Google Scholar]
- 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]
- Buratowski S., Hahn S., Sharp P. A., Guarente L. Function of a yeast TATA element-binding protein in a mammalian transcription system. Nature. 1988 Jul 7;334(6177):37–42. doi: 10.1038/334037a0. [DOI] [PubMed] [Google Scholar]
- Butt T. R., Ecker D. J. Yeast metallothionein and applications in biotechnology. Microbiol Rev. 1987 Sep;51(3):351–364. doi: 10.1128/mr.51.3.351-364.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Butt T. R., Jonnalagadda S., Monia B. P., Sternberg E. J., Marsh J. A., Stadel J. M., Ecker D. J., Crooke S. T. Ubiquitin fusion augments the yield of cloned gene products in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2540–2544. doi: 10.1073/pnas.86.8.2540. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Butt T. R., Khan M. I., Marsh J., Ecker D. J., Crooke S. T. Ubiquitin-metallothionein fusion protein expression in yeast. A genetic approach for analysis of ubiquitin functions. J Biol Chem. 1988 Nov 5;263(31):16364–16371. [PubMed] [Google Scholar]
- Celeste A. J., Rosen V., Buecker J. L., Kriz R., Wang E. A., Wozney J. M. Isolation of the human gene for bone gla protein utilizing mouse and rat cDNA clones. EMBO J. 1986 Aug;5(8):1885–1890. doi: 10.1002/j.1460-2075.1986.tb04440.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chodosh L. A., Olesen J., Hahn S., Baldwin A. S., Guarente L., Sharp P. A. A yeast and a human CCAAT-binding protein have heterologous subunits that are functionally interchangeable. Cell. 1988 Apr 8;53(1):25–35. doi: 10.1016/0092-8674(88)90484-9. [DOI] [PubMed] [Google Scholar]
- Dame M. C., Pierce E. A., Prahl J. M., Hayes C. E., DeLuca H. F. Monoclonal antibodies to the porcine intestinal receptor for 1,25-dihydroxyvitamin D3: interaction with distinct receptor domains. Biochemistry. 1986 Aug 12;25(16):4523–4534. doi: 10.1021/bi00364a011. [DOI] [PubMed] [Google Scholar]
- Demay M. B., Roth D. A., Kronenberg H. M. Regions of the rat osteocalcin gene which mediate the effect of 1,25-dihydroxyvitamin D3 on gene transcription. J Biol Chem. 1989 Feb 5;264(4):2279–2282. [PubMed] [Google Scholar]
- Fischer J. A., Giniger E., Maniatis T., Ptashne M. GAL4 activates transcription in Drosophila. Nature. 1988 Apr 28;332(6167):853–856. doi: 10.1038/332853a0. [DOI] [PubMed] [Google Scholar]
- Gallop P. M., Lian J. B., Hauschka P. V. Carboxylated calcium-binding proteins and vitamin K. N Engl J Med. 1980 Jun 26;302(26):1460–1466. doi: 10.1056/NEJM198006263022608. [DOI] [PubMed] [Google Scholar]
- Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
- Guarente L., Yocum R. R., Gifford P. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410–7414. doi: 10.1073/pnas.79.23.7410. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirst M. A., Hochman H. I., Feldman D. Vitamin D resistance and alopecia: a kindred with normal 1,25-dihydroxyvitamin D binding, but decreased receptor affinity for deoxyribonucleic acid. J Clin Endocrinol Metab. 1985 Mar;60(3):490–495. doi: 10.1210/jcem-60-3-490. [DOI] [PubMed] [Google Scholar]
- Kakidani H., Ptashne M. GAL4 activates gene expression in mammalian cells. Cell. 1988 Jan 29;52(2):161–167. doi: 10.1016/0092-8674(88)90504-1. [DOI] [PubMed] [Google Scholar]
- Lian J. B., Coutts M., Canalis E. Studies of hormonal regulation of osteocalcin synthesis in cultured fetal rat calvariae. J Biol Chem. 1985 Jul 25;260(15):8706–8710. [PubMed] [Google Scholar]
- Ma J., Przibilla E., Hu J., Bogorad L., Ptashne M. Yeast activators stimulate plant gene expression. Nature. 1988 Aug 18;334(6183):631–633. doi: 10.1038/334631a0. [DOI] [PubMed] [Google Scholar]
- Mangelsdorf D. J., Pike J. W., Haussler M. R. Avian and mammalian receptors for 1,25-dihydroxyvitamin D3: in vitro translation to characterize size and hormone-dependent regulation. Proc Natl Acad Sci U S A. 1987 Jan;84(2):354–358. doi: 10.1073/pnas.84.2.354. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McDonnell D. P., Mangelsdorf D. J., Pike J. W., Haussler M. R., O'Malley B. W. Molecular cloning of complementary DNA encoding the avian receptor for vitamin D. Science. 1987 Mar 6;235(4793):1214–1217. doi: 10.1126/science.3029866. [DOI] [PubMed] [Google Scholar]
- Metzger D., White J. H., Chambon P. The human oestrogen receptor functions in yeast. Nature. 1988 Jul 7;334(6177):31–36. doi: 10.1038/334031a0. [DOI] [PubMed] [Google Scholar]
- Pan L. C., Price P. A. The effect of transcriptional inhibitors on the bone gamma-carboxyglutamic acid protein response to 1,25-dihydroxyvitamin D3 in osteosarcoma cells. J Biol Chem. 1984 May 10;259(9):5844–5847. [PubMed] [Google Scholar]
- Pike J. W. Evidence for a reactive sulfhydryl in the DNA binding domain of the 1,25-dihydroxyvitamin D3 receptor. Biochem Biophys Res Commun. 1981 Jun;100(4):1713–1719. doi: 10.1016/0006-291x(81)90716-6. [DOI] [PubMed] [Google Scholar]
- Porzio M. A., Pearson A. M. Improved resolution of myofibrillar proteins with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochim Biophys Acta. 1977 Jan 25;490(1):27–34. doi: 10.1016/0005-2795(77)90102-7. [DOI] [PubMed] [Google Scholar]
- Schena M., Yamamoto K. R. Mammalian glucocorticoid receptor derivatives enhance transcription in yeast. Science. 1988 Aug 19;241(4868):965–967. doi: 10.1126/science.3043665. [DOI] [PubMed] [Google Scholar]
- Sentenac A. Eukaryotic RNA polymerases. CRC Crit Rev Biochem. 1985;18(1):31–90. doi: 10.3109/10409238509082539. [DOI] [PubMed] [Google Scholar]
- Webster N., Jin J. R., Green S., Hollis M., Chambon P. The yeast UASG is a transcriptional enhancer in human HeLa cells in the presence of the GAL4 trans-activator. Cell. 1988 Jan 29;52(2):169–178. doi: 10.1016/0092-8674(88)90505-3. [DOI] [PubMed] [Google Scholar]
- Wecksler W. R., Norman A. W. An hydroxylapatite batch assay for the quantitation of 1alpha,25-dihydroxyvitamin D3-receptor complexes. Anal Biochem. 1979 Jan 15;92(2):314–323. doi: 10.1016/0003-2697(79)90664-x. [DOI] [PubMed] [Google Scholar]
- West R. W., Jr, Yocum R. R., Ptashne M. Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG. Mol Cell Biol. 1984 Nov;4(11):2467–2478. doi: 10.1128/mcb.4.11.2467. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yoon K. G., Rutledge S. J., Buenaga R. F., Rodan G. A. Characterization of the rat osteocalcin gene: stimulation of promoter activity by 1,25-dihydroxyvitamin D3. Biochemistry. 1988 Nov 15;27(23):8521–8526. doi: 10.1021/bi00423a003. [DOI] [PubMed] [Google Scholar]
- Zedan H. H., el-Tayeb O. M., Abdel-Aziz M. Microbiological transformations of steroids. III. Transformation of progesterone to androstadienedione by the mycelia and by non-germinating spores of two fungi. Planta Med. 1976 Dec;30(4):360–368. doi: 10.1055/s-0028-1097744. [DOI] [PubMed] [Google Scholar]