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Submitted on February 8, 2002
Accepted on January 15, 2003
1 Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston TX 77030, Ph 713 798 6259, Fax 713 790 1275, email rrylr@bcm.tmc.edu; present address: Departmento de Bioquimica y Biologia Molecular, Facultad de Ciencias de la Salud, Universidad de Las Palmas Gran Canaria, 35016 Las Palmas de Gran Canaria, Spain; present address: Unidad de Genética Molecular, Hospital Marqués de Valdecilla, Edificio Escuela Universitaria de Enfermería, Avda. Valdecilla s/n, 39008 Santander, Spain
* To whom correspondence should be addressed. E-mail: rrylr{at}bcm.tmc.edu.
Early growth response factor (Egr-1) is an inducible zinc finger transcription factor that binds specific GC-rich enhancer elements and impacts female reproduction. These studies document for the first time that FSH rapidly induces Egr-1 expression in granulosa cells of small growing follicles. This response is transient but is reinitiated in preovulatory follicles exposed to the LH analog, hCG. Immunohistochemical analysis also showed gonadotropin induced Egr-1 in theca cells. The Egr-1 gene regulatory region responsive to gonadotropin signaling was localized within -164bp of the transcription initiation site. Binding of Sp1/Sp3 to a proximal GC-box at -64/-46bp was enhanced by FSH in immature granulosa cells but reduced after hCG stimulation of preovulatory follicles despite constant protein expression. This dynamic regulation of Sp1 binding was dependent on gonadotropin-regulated mechanisms that modulate Sp1/3-DNA binding activity. Serum response factor (SRF) was active in granulosa cells and bound a consensus CArG-box/SRE site while two putative cAMP response elements within the -164bp region bound CREB and a second cAMP-inducible protein immunologically related to CREB. Transient transfection analyses using Egr-1 promoter-luciferase constructs and site-specific mutations show that the SRE, GC-box and CRE-131 are involved in gonadotropin regulation of Egr-1 expression in granulosa cells. Specific kinase inhibitors of Erk or PKA antagonized this induction while exogenously expressed Egr-1 enhanced reporter expression. These observations indicate that the Egr-1 gene is a target of both FSH and LH action that may mediate molecular programs of proliferation and/or differentiation during follicle growth, ovulation and luteinization.
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