Interaction of phosphorylated c-Jun with TCF4 regulates intestinal cancer development

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Abstract

The proto-oncoprotein c-Jun is a component of the AP-1 transcription factor, the activity of which is augmented in many tumour types1. An important mechanism in the stimulation of AP-1 function is amino-terminal phosphorylation of c-Jun by the c-Jun N-terminal kinases (JNKs)2. Phosphorylated c-Jun is biologically more active, partially because it acquires the ability to interact with binding partners. Here we show that phosphorylated c-Jun interacts with the HMG-box transcription factor TCF4 to form a ternary complex containing c-Jun, TCF4 and β-catenin. Chromatin immunoprecipitation assays revealed JNK-dependent c-Jun–TCF4 interaction on the c-jun promoter, and c-Jun and TCF4 cooperatively activated the c-jun promoter in reporter assays in a β-catenin-dependent manner. In the ApcMin mouse model of intestinal cancer6, genetic abrogation of c-Jun N-terminal phosphorylation3 or gut-specific conditional c-jun inactivation4,5 reduced tumour number and size and prolonged lifespan. Therefore, the phosphorylation-dependent interaction between c-Jun and TCF4 regulates intestinal tumorigenesis by integrating JNK and APC/β-catenin, two distinct pathways activated by WNT signalling.

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Figure 1: TCF4 interacts with phosphorylated c-Jun.
Figure 2: JNK-dependent interaction between c-Jun and TCF4 regulates the c- jun promoter.
Figure 3: Absence of c-Jun N-terminal phosphorylation attenuates intestinal cancer development in Apc Min mice.
Figure 4: c-Jun is required for intestinal cancer development in Apc Min/+ mice.

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Acknowledgements

We would like to thank T. Akiyama, K. Niemann, G. Peters and F. Watt for providing essential reagents. We are grateful to A. Gilkar, W. Jochum, E. Nye and G. Stamp for technical help and advice on histology. We thank K. Sabapathy, I. Tomlinson and R. Treisman for critical reading of the manuscript. The London Research Institute is funded by CR UK.

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Correspondence to Axel Behrens.

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Competing interests

A.S.N. and A.B. are named as inventors on a patent application proposing inhibition of the c-Jun–TCF4 interaction as a therapeutic strategy for intestinal neoplastic disease.

Supplementary information

Supplementary Figures S1-S7

This file contains seven Supplementary Figures. Supplementary Figure S1: Determination of the c-Jun phosphorylation sites required for TCF4 binding. Supplementary Figure S2: Comparable expression of c-Jun and c-Jun4A in HCT 116 cells. Supplementary Figure S3: Nuclear β-catenin protein levels in several cell lines. Supplementary Figure S4: Luciferase assays showing cooperative activation of the c-jun promoter by c-Jun/TCF4 in HCT 116 colon cancer cells, but not NIH3T3 fibroblasts. Supplementary Figure S5: Luciferase assays showing cooperative activation of the c-jun promoter by c-Jun/TCF4 in SW480 colon cancer cells. Supplementary Figure S6: Quantification of tumor number in ApcMin/+;c-jun+/+, ApcMin/+; junAA/+ and ApcMin/+; junAA/AA mice. Supplementary Figure S7: Decreased BrdU incorporation in ApcMin/+; junAA/AA adenomas. The file also includes legends to accompany the above Supplementary Figures. (PDF 636 kb)

Supplementary Methods

Additional information on methods used in this study. (PDF 61 kb)

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Nateri, A., Spencer-Dene, B. & Behrens, A. Interaction of phosphorylated c-Jun with TCF4 regulates intestinal cancer development. Nature 437, 281–285 (2005) doi:10.1038/nature03914

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