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J. Biol. Chem., Vol. 280, Issue 33, 29397-29402, August 19, 2005

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A Role for Glycogen Synthase Kinase-3 in the Mammalian Circadian Clock^*

Chisato Iitaka, Koyomi Miyazaki, Toshihiro Akaike, and Norio Ishida¶||

From the Clock Cell Biology Research Group, Institute for Biological Resource and Function, National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1, Higashi, Tsukuba, Ibaraki 305-8566, Japan, the Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-Cho, Midori-Ku, Yokohama, Kanagawa 226-8501, Japan, and the ^¶Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305-8502, Japan

The Drosophila shaggy gene product is a mammalian glycogen synthase kinase-3 (GSK-3) homologue that contributes to the circadian clock of the Drosophila through TIMELESS phosphorylation, and it regulates nuclear translocation of the PERIOD/TIMELESS heterodimer. We found that mammalian GSK-3 is expressed in the suprachiasmatic nucleus and liver of mice and that GSK-3 phosphorylation exhibits robust circadian oscillation. Rhythmic GSK-3 phosphorylation is also observed in serum-shocked NIH3T3 cells. Exposing serum-shocked NIH3T3 cells to lithium chloride, a specific inhibitor of GSK-3, increases GSK-3 phosphorylation and delays the phase of rhythmic clock gene expression. On the other hand, GSK-3 overexpression advances the phase of clock gene expression. We also found that GSK-3 interacts with PERIOD2 (PER2) in vitro and in vivo. Recombinant GSK-3 can phosphorylate PER2 in vitro. GSK-3 promotes the nuclear translocation of PER2 in COS1 cells. The present data suggest that GSK-3 plays important roles in mammalian circadian clock.

Received for publication, March 31, 2005 , and in revised form, June 17, 2005.

^* This work was supported by a grant for young researchers with term (to K. M.) from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), the Industrial Technology Research Grant Program from the New Energy and Industrial Technology Development Organization (NEDO) of Japan, an operational subsidy from the National Institute of Advanced Industrial Science and Technology (AIST), Ministry of Economy, Trade, and Industry (METI), and a grant for Clock Cell Biology region from MEXT. 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–3.

^|| To whom correspondence should be addressed. Tel.: 81-29-861-6053; Fax: 81-29-861-9499; E-mail: n.ishida{at}aist.go.jp.

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