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J. Biol. Chem., Vol. 282, Issue 14, 10553-10560, April 6, 2007

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SOX9 Regulates Prostaglandin D Synthase Gene Transcription in Vivo to Ensure Testis Development^*

Dagmar Wilhelm¹, Ryuji Hiramatsu, Hirofumi Mizusaki, Laura Widjaja, Alexander N. Combes, Yoshiakira Kanai, and Peter Koopman, An Australian Professorial Research Fellow of the Australian Research Council. Supported by grants from the Australian Research Council and the National Health and Medical Research Council (Australia)^¶2

From the Division of Molecular Genetics and Development, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Qld 4072, Australia, the Department of Veterinary Anatomy, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan, and the ^¶ARC Centre of Excellence in Biotechnology and Development, Institute for Molecular Bioscience, The University of Queensland, Brisbane, Qld 4072, Australia

In mammals, male sex is determined by the Y-chromosomal gene Sry (sex-determining region of Y chromosome). The expression of Sry and subsequently Sox9 (SRY box containing gene 9) in precursors of the supporting cell lineage results in the differentiation of these cells into Sertoli cells. Sertoli cells in turn orchestrate the development of all other male-specific cell types. To ensure that Sertoli cells differentiate in sufficient numbers to induce normal testis development, the early testis produces prostaglandin D₂ (PGD₂), which recruits cells of the supporting cell lineage to a Sertoli cell fate. Here we show that the gene encoding prostaglandin D synthase (Pgds), the enzyme that produces PGD₂, is expressed in Sertoli cells immediately after the onset of Sox9 expression. Promoter analysis in silico and in vitro identified a paired SOX/SRY binding site. Interestingly, only SOX9, and not SRY, was able to bind as a dimer to this site and transactivate the Pgds promoter. In line with this, a transgenic mouse model showed that Pgds expression is not affected by ectopic Sry expression. Finally, chromatin immunoprecipitation proved that SOX9 but not SRY binds to the Pgds promoter in vivo.

Received for publication, October 11, 2006 , and in revised form, January 22, 2007.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by a grant from the National Institutes of Health.

² To whom correspondence should be addressed. Tel.: 61-7-3346-2059; Fax: 61-7-3346-2101; E-mail: p.koopman{at}imb.uq.edu.Au.

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