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Submitted on April 9, 2002
Accepted on May 16, 2002
1 Laboratory of Molecular Dynamics, Inc., Department of Cell Biology and Anatomy, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA. Department of Medicine, University of Colorado Health Sciences Center and Research Service, Veterans Affairs Medical Center, Denver, CO 80220, USA. Department of Biochemistry, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA. Current address: Department of Molecular Embryology, Pasteur Institute, 25 rue du Dr Roux, Paris 75724, France.
* To whom correspondence should be addressed. E-mail: boockfor{at}musc.edu.
Recent reports demonstrate that the rat GnRH (GnRH) promoter is activated in an episodic fashion in immortalized GnRH neurons, but little information is available on molecular processes that contribute to this phenomenon. In this study, we dissected the regions of the rGnRH promoter that mediate these effects by testing a series of 5' deletion luciferase reporter constructs on the pattern of photonic emissions from single, living GT1--7 GnRH neuronal cells. Deletion analysis revealed that the region -2012/-1597 that contains the neuron-specific enhancer (NSE) was required for the elaboration of pulses of GnRH promoter activity. The importance of this region was supported by observations that episodic reporter activity could be transferred to a neutral non-pulsatile promoter (Rous sarcoma virus, RSV180). Immunoneutralization of Oct-1 as well as mutation of an octamer binding site located at -1787/-1783 (AT-a site) blocked the pulsatile GnRH promoter activity in GT1--7 neuronal cells. Taken together, our findings indicate that episodic GnRH gene expression is a promoter-dependent phenomenon, which is mediated by Oct-1 interaction with regulatory elements in the NSE region.
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