Molecular Cloning of LIM Homeodomain Transcription Factor Lhx2 as a Transcription Factor of Porcine Follicle-Stimulating Hormone Beta Subunit (FSHβ) Gene
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概要
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Abstract. We cloned the LIM-homeodomain protein LHX2 as a transcription factor for the porcine follicle-stimulating hormone β subunit gene (Fshβ) by the Yeast One-Hybrid Cloning System using the upstream region of -852/-746 bases (b) from the transcription start site, called Fd2, as a bait sequence. The reporter assay in LβT2 and CHO cells revealed the presence of an LHX2-responsive region other than Fd2. A potential LHX2 binding sequence was confirmed as AATTAAT containing a consensus homeodomain binding core sequence AATT by Systematic Evolution of Ligands by Exponential Enrichment analysis. DNase I footprinting demonstrated three AATTAAT sequences located at regions -835/-829, -818/-812 and -806/-800 b in the Fd2 region and 12 binding sites in the distal and proximal regions mostly containing an AATT-core sequence. RT-PCR analysis of Lhx2 expression during porcine fetal and postnatal pituitary development showed a gradual increase from fetal day (f) 40 to postnatal day (p) 8 followed by a slight decrease to p230, suggesting that LHX2 may play its role largely in the late fetal and postnatal periods. The analyses of Lhx2 expression in pituitary tumor-derived cell lines showed their expressions in cell lines including αT31, LβT2 and others. Since LHX2 was previously identified as a transcription factor for Cga and the in vitro experiments in the present study suggested that LHX2 regulated the expression of Fshβ, it is possible that LHX2 controls the synthesis of FSH at the transcription level.
著者
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SUSA Takao
Laboratory of Molecular Biology and Gene Regulation, Graduate School of Agriculture, Meiji Universit
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Susa Takao
Division Of Life Science Graduate School Of Agriculture Meiji University
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Sato T
Inst. Of Reproduction And Endocrinology Meiji Univ.
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Kato Takako
Institute Of Plasma Physics Nagoya University
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Kato Yukio
Institute Of Molecular And Cellular Regulation Gunma University
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Ishikawa Akio
Institute For Solid State Physics University Of Tokyo
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YOSHIDA Saishu
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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SANO Yoshiya
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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KITAHARA Kousuke
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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NAKAYAMA Michie
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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SUSA Takao
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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Kitahara Kousuke
Laboratory Of Molecular Biology And Gene Regulation Division Of Life Science Graduate School Of Agriculture Meiji University
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Nakayama Michie
Laboratory Of Molecular Biology And Gene Regulation Division Of Life Science Graduate School Of Agriculture Meiji University
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Ishikawa Akio
Laboratory Of Molecular Biology And Gene Regulation Division Of Life Science Graduate School Of Agriculture Meiji University
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YOSHIDA Saishu
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University
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SANO Yoshiya
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University
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SANO Yoshiya
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
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SUSA Takao
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
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KITAHARA Kousuke
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
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KATO Takako
Institute of Reproduction and Endocrinology, Meiji University, Kanagawa 214-8571, Japan
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NAKAYAMA Michie
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
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YOSHIDA Saishu
Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan
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