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Cloning of the Ovine Estrogen Receptor- Promoter and Functional Regulation by Ovine Interferon-¹

Center for Animal Biotechnology and Genomics and Department of Animal Science, Texas A&M University, College Station, Texas 77843-2471

Address all correspondence and requests for reprints to: Dr. Fuller W. Bazer, Center for Animal Biotechnology and Genomics, 2471 TAMU, Texas A&M University, College Station, Texas 77843-2471. E-mail: fbazer{at}cvm.tamu.edu

Interferon- (IFN), the ruminant pregnancy recognition signal, inhibits transcription of the estrogen receptor (ER) gene in the endometrial lumenal epithelium of the sheep uterus, thereby abrogating production of luteolytic PGF₂ pulses. The effects of IFN are mediated in part by IFN-stimulated response elements (ISREs) and IFN regulatory factor elements (IRFEs). The promoter/enhancer region of the ovine ER gene was cloned, sequenced, and predicted to contain four IRFEs and one ISRE. Electrophoretic mobility shift assays indicated that the -2110 IRFE bound only IRF-1, whereas the -1877 IRFE and the -1284 ISRE were functional in binding IRF-1 and IRF-2. IFN inhibited transcriptional activity of the 2.7-kb ovine ER promoter in transfection assays using ovine lumenal epithelium cells. Analyses of sequential 5'-deletion mutants of the ovine ER promoter indicated that the effects of IFN may be mediated by IRFEs as well as other elements. Overexpression of ovine IRF-2, but not IRF-1, inhibited transcriptional activity of several regions of the ovine ER promoter containing an IRFE or an ISRE as well as some, but not all, regions lacking these elements.

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