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J. Biol. Chem., Vol. 279, Issue 21, 22306-22313, May 21, 2004
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DNA Methylation-mediated Control of Sry Gene Expression in Mouse Gonadal Development*
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From the
Laboratory of Cellular Biochemistry, Department of Animal Resource Sciences/Veterinary Medical Sciences, The University of Tokyo, Yayoi 1-1-1, Bukyo-ku, Tokyo 113-8657, Bio-oriented Technology Research Advancement Institution, 1-40-2, Saitama-shi, Saitama 331-8537, and the ¶Division of Applied Genetics, National Institute of Genetics, Yata 1111, Mishima, Shizuoka 411-8540, Japan
DNA methylation at CpG sequences is involved in tissue-specific and developmentally regulated gene expression. The Sry (sex-determining region on the Y chromosome) gene encodes a master protein for initiating testis differentiation in mammals, and its expression is restricted to gonadal somatic cells at 10.5-12.5 days post-coitum (dpc) in the mouse. We found that in vitro methylation of the 5'-flanking region of the Sry gene caused suppression of reporter activity, implying that Sry gene expression could be regulated by DNA methylation-mediated gene silencing. Bisulfite restriction mapping and sodium bisulfite sequencing revealed that the 5'-flanking region of the Sry gene was hypermethylated in the 8.5-dpc embryos in which the Sry gene was not expressed. Importantly, this region was specifically hypomethylated in the gonad at 11.5 dpc, while the hypermethylated status was maintained in tissues that do not express the Sry gene. We concluded that expression of the Sry gene is under the control of an epigenetic mechanism mediated by DNA methylation.
Received for publication, August 27, 2003 , and in revised form, February 20, 2004.
* This work was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN). 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.
|| To whom correspondence should be addressed. Tel.: 81-3-5841-5472; Fax: 81-3-5841-8189; E-mail: ashiota{at}mail.ecc.u-tokyo.ac.jp.
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