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Submitted on May 20, 2005
Accepted on October 28, 2005
Subunit Gene by Inducible cAMP Early Repressor
Department of Biochemistry, Molecular Biology and Cell Biology, and Center for Reproductive, Science, Northwestern University, Evanston, IL 60208; Reproductive Endocrine Unit, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114
* To whom correspondence should be addressed. E-mail: k-mayo{at}northwestern.edu.
The rodent ovary is regulated throughout the reproductive cycle to maintain normal cyclicity. Ovarian follicular development is controlled by changes in gene expression in response to the gonadotropins FSH (FSH) and LH (LH). The inhibin 
subunit gene belongs to a group of genes that is positively regulated by FSH and negatively regulated by LH. Previous studies established an important role for inducible cAMP early repressor (ICER) in repression of -inhibin. These current studies investigate the mechanisms of repression by ICER. It is not clear if all four ICER isoforms expressed in the ovary can act as repressors of the inhibin subunit gene. Electrophoretic mobility shift assays (EMSAs) demonstrate binding of all isoforms to the inhibin subunit CRE, and transfection studies demonstrate that all isoforms can repress the inhibin subunit gene. Repression by ICER is dependent on its binding to DNA as demonstrated by mutations to ICER's DNA binding domain. These mutational studies also demonstrate that repression by ICER is not dependent on heterodimerization with CREB. Competitive EMSAs show that ICER effectively competes with CREB for binding to the inhibin CRE in vitro. Chromatin immunoprecipitation (ChIP) assays demonstrate a replacement of CREB dimers bound to the inhibin CRE by ICER dimers in ovarian granulosa cells in response to LH signaling. Thus, there is a temporal association of transcription factors bound to the inhibin -CRE controlling inhibin subunit gene expression.
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