Cloning of the ovine estrogen receptor-alpha promoter and functional regulation by ovine interferon-tau.
Interferon-tau (IFNtau), the ruminant pregnancy recognition signal, inhibits transcription of the estrogen receptor alpha (ERalpha) gene in the endometrial lumenal epithelium of the sheep uterus, thereby abrogating production of luteolytic PGF(2alpha) pulses. The effects of IFNtau are mediated in part by IFN-stimulated response elements (ISREs) and IFN regulatory factor elements (IRFEs). The promoter/enhancer region of the ovine ERalpha 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. IFNtau inhibited transcriptional activity of the 2.7-kb ovine ERalpha promoter in transfection assays using ovine lumenal epithelium cells. Analyses of sequential 5'-deletion mutants of the ovine ERalpha promoter indicated that the effects of IFNtau 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 ERalpha promoter containing an IRFE or an ISRE as well as some, but not all, regions lacking these elements.
Fleming
J A
JA
Center for Animal Biotechnology and Genomics and Department of Animal Science, Texas A&M University, College Station, Texas 77843-2471, USA.
Choi
Y
Y
Johnson
G A
GA
Spencer
T E
TE
Bazer
F W
FW
D000818:Animals; D019677:Artificial Gene Fusion / Q000187:drug effects; D001483:Base Sequence / Q000235:genetics; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D004586:Electrophoresis; D004717:Endometrium / Q000166:cytology / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism; D047628:Estrogen Receptor alpha; D005260:Female; D017353:Gene Deletion; D050836:Interferon Regulatory Factor-2; D007370:Interferon Type I / Q000494:pharmacology / Q000502:physiology*; D009154:Mutation; D011257:Pregnancy Proteins / Q000494:pharmacology / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D011960:Receptors, Estrogen / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D012097:Repressor Proteins; D012756:Sheep / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000187:drug effects
D004268:DNA-Binding Proteins; D047628:Estrogen Receptor alpha; D050836:Interferon Regulatory Factor-2; D007370:Interferon Type I; D011257:Pregnancy Proteins; D011960:Receptors, Estrogen; D011994:Recombinant Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C020947:interferon tau; D013937:Thymidine Kinase