Gonadotropin-releasing hormone (GnRH) I and II are hypothalamic decapeptides with pivotal roles in the development of reproductive competence and regulation of reproductive events. In this study, transcriptional regulation of the human GnRH II gene was investigated. By scanning mutation analysis coupled with transient promoter assays, the motif at -641/-636 (CATGCC, designated GII-Sil) was identified as a repressor element. Mutation of this motif led to full restoration of promoter activity in TE671 medulloblastoma and JEG-3 placenta choriocarcinoma cells. Supershift and chromatin immunoprecipitation assays showed in vitro and in vivo binding of NF-kappa B subunit p65 and the retinoic acid receptors, RAR alpha and RXR alpha, to the promoter sequences. Over-expression of these protein factors indicated that p65 is a potent repressor, and the RAR alpha/RXR alpha heterodimer is involved in the differential regulation of the GnRH II gene in neuronal and placental cells. This was confirmed by quantitative real-time PCR. Treatment of cells with the RAR alpha/RXR alpha ligands, all-trans retinoic acid and 9-cis-retinoic acid, reduced and increased GnRH II gene expression in TE671 and JEG-3 cells, respectively. Taken together, these data demonstrate the differential roles of NF-kappa B p65 and RAR alpha/RXR alpha, interacting with the same sequence in the promoter of the human GnRH II gene to influence gene expression in a cell-specific manner.
机构:
Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Kong, Hee Jeong
Nam, Bo-Hye
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Nam, Bo-Hye
Kim, Young-Ok
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Kim, Young-Ok
Kim, Woo-Jin
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Kim, Woo-Jin
Cho, Hyun Kook
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Cho, Hyun Kook
Lee, Chang Hoon
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Pusan Natl Univ, Res Grp Marine & Silver Biotechnol, Pusan 609735, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Lee, Chang Hoon
Lee, Sang-Jun
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
Lee, Sang-Jun
Kim, Kyung-Kil
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Natl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South KoreaNatl Fisheries Res & Dev Inst, Biotechnol Res Div, Pusan 619705, South Korea
机构:
Univ Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USAUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Sun, Shan
Guo, Mian
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Univ Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Harbin Med Univ, Affiliated Hosp 2, Dept Neurosurg, Harbin, Peoples R ChinaUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Guo, Mian
Zhang, James Beiji
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Univ Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USAUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Zhang, James Beiji
Ha, Albert
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Univ Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USAUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Ha, Albert
Yokoyama, Kazunari K.
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Kaohsiung Med Univ, Grad Inst Med, Kaohsiung, TaiwanUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Yokoyama, Kazunari K.
Chiu, Robert H.
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Univ Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA
Univ Calif Los Angeles, Sch Dent, Los Angeles, CA 90024 USA
Univ Calif Los Angeles, Surg Oncol & Jonsson Comprehens Canc Ctr, Los Angeles, CA USAUniv Calif Los Angeles, Dent & Craniofacial Res Inst, Los Angeles, CA 90095 USA