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Originally published In Press as doi:10.1074/jbc.M110248200 on November 15, 2001

J. Biol. Chem., Vol. 277, Issue 3, 1884-1891, January 18, 2002
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The Transcriptional Factor Tcf-4 Contains Different Binding Sites for beta -Catenin and Plakoglobin*

Susana MiravetDagger §, José PiedraDagger §, Francesc Miró||, Emilio Itarte||, Antonio García de Herreros**, and Mireia DuñachDagger **

From the Dagger  Unitat de Biofísica, Departament de Bioquímica i Biologia Molecular, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain, || Unitat de Bioquímica de Ciències, Departament de Bioquímica i Biologia Molecular, Facultat de Ciències, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain, and  Unitat de Biologia Cellular i Molecular, Institut Municipal d'Investigació Mèdica, Universitat Pompeu Fabra, 08003 Barcelona, Spain

beta -Catenin and plakoglobin are two related armadillo proteins necessary for the establishment of adhesion junctions and desmosomes. Moreover, beta -catenin can also act as a transcriptional co-activator through its interaction with the members of Tcf/LEF-1 transcriptional factor family. We show here that Tcf-4 can be phosphorylated in vitro by protein kinase CK2 stoichiometrically in amino acids Ser-58-Ser-59-Ser-60. Phosphorylation of these residues does not modify the interaction of Tcf-4 with beta -catenin but reduces its association to plakoglobin. The binding sites of Tcf-4 for these two proteins were compared; whereas beta -catenin requires the N-terminal first 50 amino acids, plakoglobin interacts mainly with residues 51-80. Tcf-4-(51-80) binds plakoglobin in the region of armadillo repeats 1-6. Ternary complexes composed by beta -catenin/Tcf-4/plakoglobin could be detected in vitro, demonstrating that simultaneous binding of the two armadillo proteins to Tcf-4 is possible. Experiments performed using a Tcf-4 mutant with decreased interaction to plakoglobin demonstrated that binding to this protein negatively affected the transcriptional activity of Tcf-4. These results indicate that Tcf-4 contains two different sites for binding of beta -catenin and plakoglobin, and the interaction of the latter hinders the transcriptional activity of the complex.

* This work was supported by Ministerio de Ciencia y Tecnología Grants PM99-0064 and PM99-0132 (to M. D. and A. G. H., respectively), La Marató de TV3 Grant 983110 (to A. G. H.), FEDER-Fondo Nacional I+D Funds Grants 2FD97-1491-C02-02 and 2FD97-1491-C02-01 (from) (to M. D. and A. G. H., respectively), and Dirección General de Investigación Científica y Técnica Grant PB98-0856 (to E. I.).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.

§ Recipients of predoctoral fellowships from the Universitat Autònoma de Barcelona and the Ministerio de Educación, respectively.

** To whom correspondence may be addressed. Tel.: 34-93-581-1870; Fax: 34-93-581-1907; E-mail: mireia.dunach@uab.es (to M. Duñach) or Institut Municipal d'Investigació Mèdica, c/Dr. Aiguader 80, 08003 Barcelona, Spain. Tel.: 34-93-221-1009; Fax: 34-93-221-3237; E-mail: agarcia@imim.es (to A. G. de Herreros).

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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