Cloning and Characterization of Porcine CArG Binding Factor A Expression in the Anterior Pituitary

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概要

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• CArG binding factor A (CBF-A) is a transcription factor first isolated from mouse C2 myogenic cells. Several lines of evidence indicate that CBF-A is also present in the anterior pituitary lobe and participates in the process of development and cell transformation. This study was performed to clone porcine CBF-A and to investigate its roles in the porcine anterior pituitary lobe. The predicted amino acid sequence of porcine CBF-A showed a unique insertion of five TG-repeats in the N-terminal region in comparison with those of other mammals, whereas the other regions appeared to be mostly conserved including two RNA recognition motifs in the middle region. Investigation of the expression of CBF-A gene during porcine pituitary development by RT-PCR showed an exclusive and temporary decrease in expression level shortly after birth in both sexes that was gradually but insufficiently restored. The expression of fluorescence protein-fused CBF-A in CHO cells demonstrated that CBF-A is located in the nuclei. We examined whether CBF-A regulates the expression of pituitary hormone genes in CHO cells and found that CBF-A significantly stimulated the promoter activity of growth hormone and prolactin by about 2-fold but did not stimulate the LHβ gene. The specific DNA binding ability of porcine CBF-A was examined using serial oligonucleotides, CArG box and CC(W)0-6GG (W=A or T). As a result, porcine CBF-A was shown to have a high binding affinity for double- and single-stranded CC(W)6GG but no affinity for the known sequences of the CBF-A-target genes. Accordingly, this study demonstrated that porcine CBF-A may play a role in regulating at least two pituitary hormone genes, GH and PRL.

• 日本繁殖生物学会の論文
• 2008-12-01

著者

• Ishikawa Akio

  Division Of Life Science Graduate School Of Agriculture Meiji University

• Kato T

  Institute Of Reproduction And Endocrinology Meiji University

• KATO Takako

  Institute of Reproduction and Endocrinology, Meiji University

• TAKAHASHI Jun

  Laboratory of Molecular Biology and Gene Regulation, Graduate School of Agriculture, Meiji Universit

• ISHIKAWA Akio

  Laboratory of Molecular Biology and Gene Regulation, Graduate School of Agriculture, Meiji Universit

• SUSA Takao

  Laboratory of Molecular Biology and Gene Regulation, Graduate School of Agriculture, Meiji Universit

• KATO Yukio

  Laboratory of Molecular Biology and Gene Regulation, Graduate School of Agriculture, Meiji Universit

• Sato Takanobu

  Lab. Of Molecular Biology And Gene Regulation Graduate School Of Agriculture

• Susa Takao

  明治大学 農学部生命科学科

• Kato Yukio

  Department Of Dental And Medical Biochemistry Hiroshima University Graduate School Of Biomedical Sci

• Kato Y

  Meiji Univ. Kanagawa Jpn

• Susa Takao

  Division Of Life Science Graduate School Of Agriculture Meiji University

• Kato T

  Waseda Univ. Tokyo Jpn

• Sato T

  Inst. Of Reproduction And Endocrinology Meiji Univ.

• Kato Takako

  Institute Of Reproduction And Endocrinology Meiji University

• Kato Yukio

  Department Of Dental And Medical Biochemistry Hiroshima University Graduate School Of Biomedical Sci

• Kato Takako

  Institute Of Plasma Physics Nagoya University

• Kato Y

  Department Of Dental And Medical Biochemistry Hiroshima University Graduate School Of Biomedical Sci

• Kato Yukio

  Laboratory Of Molecular Biology And Gene Regulation Department Of Life Science Faculty Of Agricultur

• Takahashi Jun

  Laboratory Of Molecular Biology And Gene Regulation Graduate School Of Agriculture Meiji University

• Ishikawa Akio

  Laboratory Of Molecular Biology And Gene Regulation Division Of Life Science Graduate School Of Agriculture Meiji University

• SUSA Takao

  Laboratory of Molecular Biology and Gene Regulation, Division of Life Science, Graduate School of Agriculture, Meiji University, Kanagawa 214-8571, Japan

• SUSA Takao

  Laboratory of Molecular Biology and Gene Regulation, Department of Life Science, School of Agriculture, Meiji University

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