CpG methylation at the USF-binding site is important for the liver-specific transcription of the chipmunk HP-27 gene

被引:46
作者
Fujii, G [1 ]
Nakamura, Y [1 ]
Tsukamoto, D [1 ]
Ito, M [1 ]
Shiba, T [1 ]
Takamatsu, N [1 ]
机构
[1] Kitasato Univ, Sch Sci, Dept Biosci, Sagamihara, Kanagawa 2288555, Japan
关键词
chipmunk; CpG methylation; hibernation; HP-27; gene; tissue-specific transcription; upstream stimulatory factor (USF);
D O I
10.1042/BJ20051802
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:203 / 209
页数:7
相关论文
共 40 条
[1]   Low-temperature carbon utilization is regulated by novel gene activity in the heart of a hibernating mammal [J].
Andrews, MT ;
Squire, TL ;
Bowen, CM ;
Rollins, MB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (14) :8392-8397
[2]  
BEARD C, 1995, CELL, V65, P2325
[3]   Methylation-induced repression - Belts, braces, and chromatin [J].
Bird, AP ;
Wolffe, AP .
CELL, 1999, 99 (05) :451-454
[4]   CPG ISLANDS AS GENE MARKERS IN THE VERTEBRATE NUCLEUS [J].
BIRD, AP .
TRENDS IN GENETICS, 1987, 3 (12) :342-347
[5]   GENE NUMBER, NOISE-REDUCTION AND BIOLOGICAL COMPLEXITY [J].
BIRD, AP .
TRENDS IN GENETICS, 1995, 11 (03) :94-100
[6]   Differential regulation of uncoupling protein gene homologues in multiple tissues of hibernating ground squirrels [J].
Boyer, BB ;
Barnes, BM ;
Lowell, BB ;
Grujic, D .
AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY, 1998, 275 (04) :R1232-R1238
[7]   CPG methylation as a mechanism for the regulation of E2F activity [J].
Campanero, MR ;
Armstrong, MI ;
Flemington, EK .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2000, 97 (12) :6481-6486
[8]   Cytosine methylation of an Sp1 site contributes to organ-specific and cell-specific regulation of expression of the lung epithelial gene T1α [J].
Cao, YX ;
Jean, JC ;
Williams, MC .
BIOCHEMICAL JOURNAL, 2000, 350 :883-890
[9]   DNA methylation regulates placental lactogen I gene expression [J].
Cho, JH ;
Kimura, H ;
Minami, T ;
Ohgane, J ;
Hattori, N ;
Tanaka, S ;
Shiota, K .
ENDOCRINOLOGY, 2001, 142 (08) :3389-3396
[10]   CPG METHYLATION INHIBITS PROENKEPHALIN GENE-EXPRESSION AND BINDING OF THE TRANSCRIPTION FACTOR AP-2 [J].
COMB, M ;
GOODMAN, HM .
NUCLEIC ACIDS RESEARCH, 1990, 18 (13) :3975-3982