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Submitted on September 5, 2002
Accepted on January 7, 2003
1 Department of Animal Science, Department of Veterinary Anatomy and Public Health, Faculty of Genetics, Texas A&M University; College Station, TX 77843-2471, Phone 979-862-2790, FAX 979-862-3399, e-mail: ning@cvm.tamu.edu
* To whom correspondence should be addressed.
The preovulatory surge of estrogen up-regulates estrogen receptor-
(ER) gene expression in the uterus during the estrous/menstrual cycles of female mammals. Previously, we demonstrated that the five-fold increase in ER mRNA levels in endometrium of ovariectomized ewes treated with a physiological dose of estradiol (E2) is entirely due to an increase in ER mRNA stability. Our current work confirms that the E2 effect is specific to ER mRNA. The sequence of ER mRNA, cloned from sheep endometrium, shows a high degree of conservation with those of other species, even in the 5' and the very long 3' untranslated regions (UTRs). In a cell-free assay, ER mRNA demonstrates greater stability with endometrial extracts from E2-treated ewes compared with those from untreated ovariectomized ewes. The E2-enhanced stability of ER mRNA was ablated by prior treatment of the extracts with proteinase K, 70 C heat, oxidizing and alkylating reagents, indicating that a protein is responsible for stabilization of the message. The 3'UTR of ER mRNA contains discrete sequences required for E2-enhanced stability, four of which were identified by extensive deletion mutant analyses. Transfer of two of the four "Minimal E2-Modulated Stability Sequences" (MEMSS) conferred E2-enhanced stability to a heterologous RNA. These MEMSS contain a common 10 base, uridine-rich sequence that is predicted to reside in a loop structure. Throughout our studies, estrogen stabilization of ER mRNA in sheep endometrium resembled that of vitellogenin mRNA in frog liver, indicating conservation of this ancient mechanism for enhancing gene expression in response to estrogen.
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