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Department of Molecular and Integrative Physiology University of Kansas Medical Center Kansas City, Kansas 66160-7401
The hypothalamic neuropeptide, GnRH, regulates the
synthesis and secretion of LH from pituitary gonadotropes. Furthermore,
it has been shown that the LH ß-subunit gene is regulated by the
transcription factors steroidogenic factor-1 (SF-1) and early growth
response protein 1 (Egr1) in vitro and in vivo.
The present study investigated the roles played by Egr1 and SF-1 in
regulating activity of the equine LHß-subunit promoter in the
gonadotrope cell line, 
T3–1, and the importance of these factors
and cis-acting elements in regulation of the promoter by
GnRH. All four members of the Egr family were found to induce activity
of the equine promoter. The region responsible for induction by Egr was
localized to the proximal 185 bp of the promoter, which contained two
Egr response elements. Coexpression of Egr1 and SF-1 led to a
synergistic activation of the equine (e)LHß promoter. Mutation of any
of the Egr or SF-1 response elements attenuated this synergism.
Endogenous expression of Egr1 in T3–1 cells was not detectable
under basal conditions, but was rapidly induced after GnRH stimulation.
Reexamination of the promoter constructs harboring mutant Egr or SF-1
sites indicated that these sites were required for GnRH induction. In
fact, mutation of both Egr sites within the eLHß promoter completely
attenuated its induction by GnRH. Thus, GnRH induces expression of
Egr1, which subsequently activates the eLHß promoter. Finally, GnRH
not only induced expression of Egr1, but also its corepressor, NGFI-A
(Egr1) binding protein (Nab1), which can repress Egr1- induced
transcription of the eLHß promoter.
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