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Nucleic Acids Research 2005 33(10):3401-3411; doi:10.1093/nar/gki652
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Published online 10 June 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

cAMP-responsive element in TATA-less core promoter is essential for haploid-specific gene expression in mouse testis

Pranee Somboonthum, Hiroshi Ohta, Shuichi Yamada1, Masayoshi Onishi, Akiko Ike, Yoshitake Nishimune and Masami Nozaki*

Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University Suita, Osaka 565-0871, Japan 1Department of Cell Biology, Institute for Virus Research, Kyoto University Kyoto 606-8507, Japan

*To whom correspondence should be addressed. Tel/Fax: +816 6879 8339; Email: mnozaki{at}biken.osaka-u.ac.jp

Received April 11, 2005. Revised May 23, 2005. Accepted May 23, 2005.

Promoters, including neither TATA box nor initiator, have been frequently found in testicular germ cell-specific genes in mice. These investigations imply that unique forms of the polymerase II transcription initiation machinery play a role in selective activation of germ cell-specific gene expression programs during spermatogenesis. However, there is little information about testis-specific core promoters, because useful germ cell culture system is not available. In this study, we characterize the regulatory region of the haploid-specific Oxct2b gene in detail by using in vivo transient transfection assay in combination with a transgenic approach, with electrophoretic mobility shift and chromatin immunoprecipitation assays. Expression studies using mutant constructs demonstrate that a 34 bp region, which extends from –49 to –16, acts as a core promoter in an orientation-dependent manner. This promoter region includes the cAMP-responsive element (CRE)-like sequence TGACGCAG, but contains no other motifs, such as a TATA box or initiator. The CRE-like element is indispensable for the core promoter activity, but not for activator in testicular germ cells, through the binding of a testis-specific CRE modulator transcription factor. These results indicate the presence of alternative transcriptional initiation machinery for cell-type-specific gene expression in testicular germ cells.

Present address: Hiroshi Ohta, Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN, Kobe, Hyogo 650-0047, Japan


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