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Originally published In Press as doi:10.1074/jbc.M212635200 on June 7, 2003

J. Biol. Chem., Vol. 278, Issue 35, 33067-33077, August 29, 2003
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Inhibitory Phosphorylation of Glycogen Synthase Kinase-3 (GSK-3) in Response to Lithium

EVIDENCE FOR AUTOREGULATION OF GSK-3*

Fang Zhang {ddagger}, Christopher J. Phiel §, Laura Spece §, Nadia Gurvich {ddagger} and Peter S. Klein § ¶

From the {ddagger}Department of Pharmacology and §Howard Hughes Medical Institute and Department of Medicine, Division of Hematology-Oncology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6148

Glycogen synthase kinase-3 (GSK-3) is a critical, negative regulator of diverse signaling pathways. Lithium is a direct inhibitor of GSK-3 and has been widely used to test the putative role of GSK-3 in multiple settings. However, lithium also inhibits other targets, including inositol monophosphatase and structurally related phosphomonoesterases, and thus additional approaches are needed to attribute a given biological effect of lithium to a specific target. For example, lithium is known to increase the inhibitory N-terminal phosphorylation of GSK-3, but the target of lithium responsible for this indirect regulation has not been identified. We have characterized a short peptide derived from the GSK-3 interaction domain of Axin that potently inhibits GSK-3 activity in vitro and in mammalian cells and robustly activates Wnt-dependent transcription, mimicking lithium action. We show here, using the GSK-3 interaction domain peptide, as well as small molecule inhibitors of GSK-3, that lithium induces GSK-3 N-terminal phosphorylation through direct inhibition of GSK-3 itself. Reduction of GSK-3 protein levels, either by RNA interference or by disruption of the mouse GSK-3{beta} gene, causes increased N-terminal phosphorylation of GSK-3, confirming that GSK-3 regulates its own phosphorylation status. Finally, evidence is presented that N-terminal phosphorylation of GSK-3 can be regulated by the GSK-3-dependent protein phosphatase-1·inhibitor-2 complex.

Received for publication, December 11, 2002 , and in revised form, May 6, 2003.

* This work was supported by a grant from the National Institute of Mental Health (to P. S. K.). 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.: 215-898-2179; Fax: 215-573-4320; E-mail: pklein{at}mail.med.upenn.edu.


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