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J. Biol. Chem., Vol. 278, Issue 37, 35819-35825, September 12, 2003

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Cellular Retinoic Acid-binding Protein II Gene Expression Is Directly Induced by Estrogen, but Not Retinoic Acid, in Rat Uterus^*

Xiao-Hong Li and David E. Ong 

From the Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee 37232

It has been suggested that cellular retinoic acid-binding protein (II) (CRABP(II)) may have a role in the movement of retinoic acid (RA) to its nuclear receptors, thereby enhancing the action of RA in the cells in which it is expressed. RA has also been shown to increase expression of CRABP(II). Previous work from our laboratory has shown that 17-estradiol (E₂) administration to prepubertal female rats leads to acquisition of the ability of the lining epithelium to synthesize RA as well as to express CRABP(II). To determine whether this appearance of CRABP(II) was dependent on the production of RA, both E₂ and RA were administered to ovariectomized rats. E₂ administration induced expression of the CRABP(II) gene in the uterus within 4 h, and this induction was not inhibited by prior administration of puromycin, indicating that the induction was direct. In contrast, RA caused no change in CRABP(II) message level, even at times as late as 48 h after administration. Isolation and analysis of 4.5 kb of the 5'-flanking region of the gene revealed no apparent E₂-response element. Using this portion of the gene to drive expression of the luciferase gene in transfected cells allowed identification of a region containing an imperfect estrogen-response element and estrogen-response element half-site, necessary for E₂-driven induction. A possible Sp1 binding site in the 5'-flanking region of the CRABP(II) gene was also required for this induction. The ability of E₂ to induce expression of CRABP(II) suggests that it can enhance the activity of RA, directly affecting expression of retinoid-responsive genes.

Received for publication, March 12, 2003 , and in revised form, June 2, 2003.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AY226560.

^* This work was supported by National Institutes of Health Grants HD25206 and DK32642. Core facilities used were from the Diabetes Center (oligonucleotide synthesis) and the Vanderbilt-Ingram Cancer Center (DNA sequencing), supported by National Institutes of Health Grants DK 20593 and CA 68485, respectively. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Biochemistry, Vanderbilt University, 23rd Ave. at Pierce, Nashville, TN 37232. Tel.: 615-322-6331; Fax: 615-343-7347; E-mail: david.e.ong{at}vanderbilt.edu.

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