Endocrinology, Vol 134, 11-18, Copyright © 1994 by Endocrine Society

ARTICLES

Calbindin-D9k gene expression in the uterus: study of the two messenger ribonucleic acid species and analysis of an imperfect estrogen- responsive element

F L'Horset, C Blin, S Colnot, M Lambert, M Thomasset and C Perret
INSERM U. 120, Hopital Robert Debre, Paris, France.

The calbindin D9k (CaBP9k) gene is under strict estrogen control in the rat uterus. This tissue contains two CaBP9k messenger RNA (mRNA) species. We have used primer extension analysis, reverse transcriptase associated with polymerase chain reaction, and RNase H digestion to show that these two mRNA species have the same structural features, including 5'- and 3'-ends, and poly(A) tail length. Our results suggest that the difference in electrophoretic mobilities of the two mRNA species might be due to interaction with another factor. We also analyzed the imperfect estrogen-responsive element (ERE) present on the first 5'-splice site of the rat CaBP9k gene. The oligonucleotide corresponding to the CaBP9k ERE was cloned in the plasmid pBLCAT2 (where the thymidine kinase promoter governs the expression of the chloramphenicol acetyl transferase gene) and transfected into MCF7 cells. This CaBP9k ERE was found to be a hormone-inducible enhancer that worked in an orientation-independent manner on a heterologous promoter and was functional at physiological hormone concentrations. One CaBP9k ERE conferred only weak (about 2-fold) estrogen induction, but two EREs cloned in tandem were strongly synergistic (14- to 16- fold). The CaBP9k ERE also bound to the partially purified estrogen receptor (ER) and to ER expressed in COS cells by gel shift assay. Methylation interference showed that all the guanine residues in both half-sites of the CaBP9k ERE were protected by ER binding. Thus, ER binds to the CaBP9k ERE in a way similar to other EREs. The gel shift assay results indicate that the strong synergistic effect of two EREs cloned in tandem is not due to cooperative binding between the two elements. As the CaBP9k gene is under strong estrogenic control in the uterus in vivo, the imperfect CaBP9k ERE may cooperate with another trans-acting factor to become fully efficient.

This article has been cited by other articles:

				V. H. Dang, T. H. Nguyen, K.-C. Choi, and E.-B. Jeung A Calcium-Binding Protein, Calbindin-D9k, Is Regulated through an Estrogen-Receptor Mediated Mechanism following Xenoestrogen Exposure in the GH3 Cell Line Toxicol. Sci., August 1, 2007; 98(2): 408 - 415. [Abstract] [Full Text] [PDF]

				V. H. Dang, K.-C. Choi, and E.-B. Jeung Tetrabromodiphenyl Ether (BDE 47) Evokes Estrogenicity and Calbindin-D9k Expression through an Estrogen Receptor-Mediated Pathway in the Uterus of Immature Rats Toxicol. Sci., June 1, 2007; 97(2): 504 - 511. [Abstract] [Full Text] [PDF]

				G.-S. Lee, K.-C. Choi, and E.-B. Jeung Glucocorticoids differentially regulate expression of duodenal and renal calbindin-D9k through glucocorticoid receptor-mediated pathway in mouse model Am J Physiol Endocrinol Metab, February 1, 2006; 290(2): E299 - E307. [Abstract] [Full Text] [PDF]

				G.-S. Lee, H.-J. Kim, Y.-W. Jung, K.-C. Choi, and E.-B. Jeung Estrogen Receptor {alpha} Pathway Is Involved in the Regulation of Calbindin-D9k in the Uterus of Immature Rats Toxicol. Sci., April 1, 2005; 84(2): 270 - 277. [Abstract] [Full Text] [PDF]

				P. V. Nantermet, P. Masarachia, M. A. Gentile, B. Pennypacker, J. Xu, D. Holder, D. Gerhold, D. Towler, A. Schmidt, D. B. Kimmel, et al. Androgenic Induction of Growth and Differentiation in the Rodent Uterus Involves the Modulation of Estrogen-Regulated Genetic Pathways Endocrinology, February 1, 2005; 146(2): 564 - 578. [Abstract] [Full Text] [PDF]

				Y.-W. Jung, E.-J. Hong, K.-C. Choi, and E.-B. Jeung Novel Progestogenic Activity of Environmental Endocrine Disruptors in the Upregulation of Calbindin-D9k in an Immature Mouse Model Toxicol. Sci., January 1, 2005; 83(1): 78 - 88. [Abstract] [Full Text] [PDF]

				G.-S. Lee, K.-C. Choi, H.-J. Kim, and E.-B. Jeung Effect of Genistein As a Selective Estrogen Receptor Beta Agonist on the Expression of Calbindin-D9k in the Uterus of Immature Rats Toxicol. Sci., December 1, 2004; 82(2): 451 - 457. [Abstract] [Full Text] [PDF]

				D. Korita, H. Itoh, N. Sagawa, S. Yura, M. Yoshida, K. Kakui, M. Takemura, and S. Fujii 17{beta}-Estradiol Up-Regulates Prostacyclin Production in Cultured Human Uterine Myometrial Cells Via Augmentation of Both Cyclooxygenase-1 and Prostacyclin Synthase Expression Reproductive Sciences, October 1, 2004; 11(7): 457 - 464. [Abstract] [PDF]

				R. O'Lone, M. C. Frith, E. K. Karlsson, and U. Hansen Genomic Targets of Nuclear Estrogen Receptors Mol. Endocrinol., August 1, 2004; 18(8): 1859 - 1875. [Abstract] [Full Text] [PDF]

				E.-J. Hong, K.-C. Choi, and E.-B. Jeung Induction of Calbindin-D9k Messenger RNA and Protein by Maternal Exposure to Alkylphenols During Late Pregnancy in Maternal and Neonatal Uteri of Rats Biol Reprod, August 1, 2004; 71(2): 669 - 675. [Abstract] [Full Text] [PDF]

				C. M. Klinge Estrogen receptor interaction with estrogen response elements Nucleic Acids Res., July 15, 2001; 29(14): 2905 - 2919. [Abstract] [Full Text] [PDF]

				S. F. Palter, A. B. Tavares, A. Hourvitz, J. D. Veldhuis, and E. Y. Adashi Are Estrogens of Import to Primate/Human Ovarian Folliculogenesis? Endocr. Rev., June 1, 2001; 22(3): 389 - 424. [Abstract] [Full Text] [PDF]

				R. A. Shapiro, C. Xu, and D. M. Dorsa Differential Transcriptional Regulation of Rat Vasopressin Gene Expression by Estrogen Receptor {alpha} and {beta} Endocrinology, November 1, 2000; 141(11): 4056 - 4064. [Abstract] [Full Text] [PDF]

				K. Tatsumi, T. Higuchi, H. Fujiwara, T. Nakayama, K. Itoh, T. Mori, S. Fujii, and J. Fujita Expression of calcium binding protein D-9k messenger RNA in the mouse uterine endometrium during implantation Mol. Hum. Reprod., February 1, 1999; 5(2): 153 - 161. [Abstract] [Full Text] [PDF]

				I. Treilleux, N. Peloux, M. Brown, and A. Sergeant Human Estrogen Receptor (ER) Gene Promoter-P1: Estradiol-Independent Activity and Estradiol Inducibility in ER+ and ER- Cells Mol. Endocrinol., August 1, 1997; 11(9): 1319 - 1331. [Abstract] [Full Text]

				T. L. Bale and D. M. Dorsa Cloning, Novel Promoter Sequence, and Estrogen Regulation of a Rat Oxytocin Receptor Gene Endocrinology, March 1, 1997; 138(3): 1151 - 1158. [Abstract] [Full Text] [PDF]

				B. Romagnolo, F. Cluzeaud, M. Lambert, S. Colnot, A. Porteu, T. Molina, M. Tomasset, A. Vandewalle, A. Kahn, and C. Perret Tissue-specific and Hormonal Regulation of Calbindin-D9K Fusion Genes in Transgenic Mice J. Biol. Chem., July 12, 1996; 271(28): 16820 - 16826. [Abstract] [Full Text] [PDF]