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Originally published In Press as doi:10.1074/jbc.M002747200 on May 11, 2000

J. Biol. Chem., Vol. 275, Issue 31, 24032-24039, August 4, 2000
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MEKK4 Mediates Differentiation in Response to Retinoic Acid via Activation of c-Jun N-terminal Kinase in Rat Embryonal Carcinoma P19 Cells*

Jyotshnabala KanungoDagger , Irina PotapovaDagger , Craig C. MalbonDagger §, and Hsien-yu Wang

From the Dagger  Department of Molecular Pharmacology, University Medical Center, SUNY/Stony Brook, Stony Brook, New York 11794-8651 and the  Department of Physiology and Biophysics, Diabetes and Metabolic Diseases Research Program, University Medical Center, SUNY/Stony Brook, Stony Brook, New York 11794-8661

Differentiation of P19 embryonal carcinoma cells in response to the morphogen retinoic acid is regulated by Galpha 12/13 and is associated with activation of c-Jun N-terminal kinase. The role of MEKK1 and MEKK4 upstream of the c-Jun N-terminal kinase was investigated in P19 cells. P19 clones stably expressing constitutively active and dominant negative mutants of MEKK1 and MEKK4 were created and characterized. Expression of the constitutively active form of either MEKK1 or MEKK4 mimicked the action of retinoic acid, inducing these embryonal carcinoma cells to primitive endoderm. Expression of the dominant negative form of MEKK1 had no influence on the ability of retinoic acid to induce either JNK activation or primitive endoderm formation in P19 stem cells. Expression of the dominant negative form of MEKK4, in contrast, effectively blocks both morphogen-induced activation of JNK and cellular differentiation. These data identify MEKK4 as upstream of c-Jun N-terminal kinase in the pathway mediating differentiation of P19 stem cells to primitive endoderm.

* This work was supported by United States Public Health Service Grant DK30111 (NIDDK, National Institutes of Health).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: Pharmacology-HSC, SUNY/Stony Brook, Stony Brook, NY 11794-8651. Tel.: 631-444-7873; Fax: 631-444-7696; E-mail: craig@pharm.som.sunysb.edu.

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