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Nucleic Acids Research 2005 33(6):1924-1934; doi:10.1093/nar/gki339
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Published online 4 April 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Article

Regulation of Id2 expression by CCAAT/enhancer binding protein ß

Kazuhiro Karaya, Seiichi Mori, Hisashi Kimoto, Yoko Shima, Yoshihito Tsuji1, Hisanori Kurooka, Shizuo Akira2 and Yoshifumi Yokota*

Department of Molecular Genetics, School of Medicine, University of Fukui 23-3 Shimoaizuki, Matsuoka, Fukui 910-1193, Japan 1Department of Neurosurgery, Kyoto University Graduate School of Medicine 54 Shogoin Kawahara-cho, Sakyo, Kyoto 606-8507, Japan 2Department of Host Defense, Research Institute for Microbial Diseases, Osaka University 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan

*To whom correspondence should be addressed. Tel: +81 776 61 8312; Fax: +81 776 61 8164; Email: yyokota{at}fmsrsa.fukui-med.ac.jp

Received March 15, 2005. Accepted March 17, 2005.

Mice deficient for Id2, a negative regulator of basic helix–loop–helix (bHLH) transcription factors, exhibit a defect in lactation due to impaired lobuloalveolar development during pregnancy, similar to the mice lacking the CCAAT enhancer binding protein (C/EBP) ß. Here, we show that Id2 is a direct target of C/EBPß. Translocation of C/EBPß into the nucleus, which was achieved by using a system utilizing the fusion protein between C/EBPß and the ligand-binding domain of the human estrogen receptor (C/EBPß-ERT), demonstrated the rapid induction of endogenous Id2 expression. In reporter assays, transactivation of the Id2 promoter by C/EBPß was observed and, among three potential C/EBPß binding sites found in the 2.3 kb Id2 promoter region, the most proximal element was responsible for the transactivation. Electrophoretic mobility shift assay (EMSA) identified this element as a core sequence to which C/EBPß binds. Chromatin immunoprecipitation (ChIP) furthermore confirmed the presence of C/EBPß in the Id2 promoter region. Northern blotting showed that Id2 expression in C/EBPß-deficient mammary glands was reduced at 10 days post coitus (d.p.c.), compared with that in wild-type mammary glands. Thus, our data demonstrate that Id2 is a direct target of C/EBPß and provide insight into molecular mechanisms underlying mammary gland development during pregnancy.


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