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Originally published In Press as doi:10.1074/jbc.C000113200 on March 15, 2000
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J Biol Chem, Vol. 275, Issue 19, 14013-14016, May 12, 2000

ACCELERATED PUBLICATION
Induction of Melanocyte-specific Microphthalmia-associated Transcription Factor by Wnt-3a*

Kazuhisa Takeda, Ken-ichi Yasumoto, Ritsuko TakadaDagger , Shinji TakadaDagger §, Ken-ichi Watanabe, Tetsuo Udono, Hideo Saito, Kazuhiro Takahashi, and Shigeki Shibahara

From the Department of Molecular Biology and Applied Physiology, Tohoku University School of Medicine, Aoba-ku, Sendai, Miyagi 980-8575, Japan, Dagger  Kondoh Differentiation Signaling Project, ERATO, Japan Science and Technology Corporation (JST), 14 Yoshidakawaramachi, Sakyo-ku, Kyoto 606-8305, Japan, and § Center for Molecular and Developmental Biology, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan

Microphthalmia-associated transcription factor (Mitf) plays a critical role in the development of neural crest-derived melanocytes. Here, we show that exogenously added Wnt-3a protein, an intercellular signaling molecule, up-regulates the expression of endogenous melanocyte-specific Mitf (Mitf-M) mRNA in cultured melanocytes. The melanocyte-specific promoter of the human MITF gene (MITF-M promoter) contains a functional LEF-1-binding site, which is bound in vitro by LEF-1 and confers the preferential expression on a reporter gene in melanocytes and melanoma cells, as judged by the transient transfection assays. Moreover, the LEF-1-binding site is required for the transactivation of a reporter gene by LEF-1, beta -catenin, or their combination. Exogenously added Wnt-3a protein also transactivates the MITF-M promoter via the LEF-1-binding site; this activation was abolished when a dominant-negative form of LEF-1 was coexpressed. These results suggest that Wnt-3a signaling recruits beta -catenin and LEF-1 to the LEF-1-binding site of the MITF-M promoter. Therefore, the present study identifies Mitf-M/MITF-M as a direct target of Wnt signaling.

* This work was supported in part by grants-in-aid for scientific research (B), for exploratory research, and for encouragement of young scientist (to K. Y.) from the Ministry of Education, Science, Sports and Culture of Japan. This work was also supported in part by the Nakatomi Foundation and the Kao Foundation for Arts and Sciences.The costs of publication of this article were defrayed in part by the payment of page charges. The 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.: 81-22-717-8117; Fax: 81-22-717-8118; E-mail: shibahar@mail.cc.tohoku.ac.jp.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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