| Project/Area Number |
13680768
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Molecular biology
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| Research Institution | 宮崎医科大学 |
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Principal Investigator |
TAKECHI Shinji University of Miyazaki, Miyazaki Medical College, Research Associate, 医学部, 助手 (10222100)
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| Co-Investigator(Kenkyū-buntansha) |
KIKUCHI Hidehiko University of Miyazaki, Frontier Science Research Center, Research Associate, フロンティア科学実験総合センター, 助手 (10301384)
NAKAYAMA Tatsuo University of Miyazaki, Frontier Science Research Center, Professor, フロンティア科学実験総合センター, 教授 (60031712)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | histone / acetylation / transcriptional control / gene expression / 脱アセチル化酵素 |
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| Research Abstract |
We have cloned the chicken homolog of OBF-1, chOBF-1, which comprises 256 amino acids, and exhibits only 65% overall identity to the human and mouse OBF-1 proteins. Amino acid sequence alignment revealed the putative Oct-binding sequence, RPYQGVRVKEPVKELL(K/R)RKRG, which is conserved among chicken, mouse and human. chOBF-1 protein was demonstrated to bind chicken Oct-1 protein by the in vitro immunoprecipitation experiment, and chOBF-1 was shown to functionally activate the chicken immunoglobulin (Ig) light chain promoter in the NIH 3T3 cell. Taken together, these data indicate that the Ig gene transcription machinery, including Oct-1 and OBF-1, has been highly conserved in vertebrate evolution.
To investigate the transcriptional function of chHDAC2, we employed the chIgM-L promoter reporter plasmid. We found that chHDAC2 represses activated chIgM-L promoter activity. In transient expression experiments in NIH 3T3 cell, the specific HDAC inhibitor tricostatin A increased transactivation of chIgM-L promoter activity mediated by chicken Oct-1 and OBF-1 proteins. In transient coexpression of the three class I chicken histone deacetylases (chHDACl-3) tested, only chHDAC2 repressed the activated chIgM-L promoter activity. These findings suggest that chHDAC2 might be recruited to the chIgM-L promoter and specifically repress chIgM-L transcription.
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