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Molecular and Cellular Biology, January 2008, p. 825-835, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.02375-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Defining the Roles of β-Catenin and Plakoglobin in LEF/T-Cell Factor-Dependent Transcription Using β-Catenin/Plakoglobin-Null F9 Cells{triangledown}

Masayuki Shimizu,1 Yoshitaka Fukunaga,1 Junichi Ikenouchi,2 and Akira Nagafuchi1*

Division of Cellular Interactions, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto 860-0811, Japan,1 Department of Cell Biology, Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan2

Received 20 December 2006/ Returned for modification 4 May 2007/ Accepted 23 October 2007

β-Catenin functions as a transcriptional regulator in Wnt signaling. Its function is regulated by a specific destruction system. Plakoglobin is a close homologue of β-catenin in mammalian cells and is regulated in a similar fashion. When β-catenin or plakoglobin is exogenously expressed in cells, endogenous β-catenin is stabilized, which complicates estimation of the transcriptional activities of exogenously expressed proteins. To facilitate the design of experiments aimed at investigating the transcriptional activities of β-catenin and plakoglobin, we utilized F9 cells in which we knocked out endogenous β-catenin and/or plakoglobin by gene deletion and exogenously expressed wild-type and mutant β-catenin and/or plakoglobin. We show that C-terminally deleted β-catenin, but not plakoglobin, has a strong dominant-negative effect on transcription without altering the nuclear accumulation of β-catenin. Moreover, we show that Wnt-3a activation of LEF/T-cell factor (TCF)-dependent transcription depends on β-catenin but not on plakoglobin. Using chimeras of β-catenin and plakoglobin, we demonstrate that plakoglobin has the potential to function in transcriptional regulation but is not responsible for Wnt-3a signaling in F9 cells. Our data show that preferential nuclear accumulation of β-catenin is not necessarily linked to its transcriptional activity. We also clearly demonstrate that plakoglobin is insufficient for LEF/TCF-dependent transcriptional activation by Wnt-3a in F9 cells.

* Corresponding author. Mailing address: Division of Cellular Interactions, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811, Japan. Phone: 81-96-373-6606. Fax: 81-96-373-6609. E-mail: naga-san{at}kumamoto-u.ac.jp

{triangledown} Published ahead of print on 5 November 2007.

Molecular and Cellular Biology, January 2008, p. 825-835, Vol. 28, No. 2
0270-7306/08/$08.00+0     doi:10.1128/MCB.02375-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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