The chipmunk hibernation-specific HP-27 gene is expressed specifically in the liver and has a CpG-poor promoter. To reveal how the liver-specific transcription of the HP-27 gene is regulated, we performed yeast one-hybrid screening of a chipmunk liver cDNA library. A 5′-flanking sequence of the HP-27 gene, extending from −170 to −140 and containing an E-box (5′-CACGTG-3′), is essential for the liver-specific transcription of HP-27. We used this sequence as bait and found that a ubiquitously expressed transcription factor, USF (upstream stimulatory factor), bound to the E-box. In COS-7 cells, USF activated transcription from the HP-27 gene promoter. We then used bisulphite genomic sequencing to analyse the methylation status of the four CpG dinucleotides that lie in the 5′-flanking sequence of the HP-27 gene up to −450, to investigate how the ubiquitously expressed USF activates transcription of the HP-27 gene only in the liver, while its transcription is repressed elsewhere. The only difference in methylation in the tissues tested was in the CpG dinucleotide in the USF-binding site, which was hypomethylated in the liver, but highly methylated in the kidney and heart. The specific methylation of the CpG dinucleotide at the USF-binding site impeded both the binding of USF and its transcriptional activation of the HP-27 gene. Chromatin immunoprecipitation using anti-USF antibodies revealed that USF bound to the HP-27 gene promoter in the liver, but not in the kidney or heart. Thus CpG methylation at the USF-binding site functions in establishing and maintaining tissue-specific transcription from the CpG-poor HP-27 gene promoter.
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Research Article|
March 15 2006
CpG methylation at the USF-binding site is important for the liver-specific transcription of the chipmunk HP-27 gene
Gen Fujii;
Gen Fujii
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
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Yuki Nakamura;
Yuki Nakamura
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
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Daisuke Tsukamoto;
Daisuke Tsukamoto
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
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Michihiko Ito;
Michihiko Ito
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
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Tadayoshi Shiba;
Tadayoshi Shiba
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
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Nobuhiko Takamatsu
Nobuhiko Takamatsu
1
1Department of Biosciences, School of Science, Kitasato University, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan
1To whom correspondence should be addressed (email takamatu@jet.sci.kitasato-u.ac.jp).
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Publisher: Portland Press Ltd
Received:
November 09 2005
Revision Received:
January 03 2006
Accepted:
January 06 2006
Accepted Manuscript online:
January 06 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 395 (1): 203–209.
Article history
Received:
November 09 2005
Revision Received:
January 03 2006
Accepted:
January 06 2006
Accepted Manuscript online:
January 06 2006
Citation
Gen Fujii, Yuki Nakamura, Daisuke Tsukamoto, Michihiko Ito, Tadayoshi Shiba, Nobuhiko Takamatsu; CpG methylation at the USF-binding site is important for the liver-specific transcription of the chipmunk HP-27 gene. Biochem J 1 April 2006; 395 (1): 203–209. doi: https://doi.org/10.1042/BJ20051802
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