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c-Jun N-terminal phosphorylation antagonises recruitment of the Mbd3/NuRD repressor complex

Nature volume 469, pages 231–235 (2011)Cite this article

Abstract

AP-1 (activator protein 1) activity is strongly induced in response to numerous signals, including growth factors, cytokines and extracellular stresses1. The proto-oncoprotein c-Jun belongs to the AP-1 group of transcription factors and it is a crucial regulator of intestinal progenitor proliferation and tumorigenesis2,3,4. An important mechanism of AP-1 stimulation is phosphorylation of c-Jun by the Jun amino-terminal kinases (JNKs)1. N-terminal phosphorylation of the c-Jun transactivation domain increases target gene transcription5,6, but a molecular explanation was elusive. Here we show that unphosphorylated, but not N-terminally phosphorylated c-Jun, interacts with Mbd3 and thereby recruits the nucleosome remodelling and histone deacetylation (NuRD) repressor complex. Mbd3 depletion in colon cancer cells increased histone acetylation at AP-1-dependent promoters, which resulted in increased target gene expression. The intestinal stem cell marker lgr5 was identified as a novel target gene controlled by c-Jun/Mbd3. Gut-specific conditional deletion of mbd3 (mbd3ΔG/ΔG mice) stimulated c-Jun activity and increased progenitor cell proliferation. In response to inflammation, mdb3 deficiency resulted in colonic hyperproliferation and mbd3ΔG/ΔG mice showed markedly increased susceptibility to colitis-induced tumorigenesis. Notably, concomitant inactivation of a single allele of c-jun reverted physiological and pathological hyperproliferation, as well as the increased tumorigenesis in mbd3ΔG/ΔG mice. Thus the transactivation domain of c-Jun recruits Mbd3/NuRD to AP-1 target genes to mediate gene repression, and this repression is relieved by JNK-mediated c-Jun N-terminal phosphorylation.

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Figure 1: Mbd3 interacts with unphosphorylated c-Jun through its MBD.
Figure 2: Mbd3 represses c-jun transcription.
Figure 3: lgr5 is regulated by c-Jun-Mbd3/NuRD.
Figure 4: Mbd3 antagonises c-Jun/AP-1 function in vivo.

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Acknowledgements

We are grateful to the LRI Animal Unit, Equipment Park, FACS, Peptide synthesis and the Experimental Histopathology unit for technical help and O. S. Gabrielsen for reagents. We thank C. Hill and H. Van Dam for critical reading of the manuscript. C.A. and R.S. were funded by Marie Curie Intraeuropean Fellowships (PIEF-GA-2008-220566 and MEIF-CT-2006-041119). The London Research Institute is funded by Cancer Research UK.

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Author notes

  1. Cristina Aguilera and Kentaro Nakagawa: These authors contributed equally to this work.

Authors and Affiliations

  1. Mammalian Genetics Laboratory, Cancer Research UK London Research Institute, Lincoln’s Inn Fields Laboratories, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK ,

    Cristina Aguilera, Kentaro Nakagawa, Rocio Sancho, Atanu Chakraborty & Axel Behrens

  2. Department of Medical Biochemistry, Graduate School of Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8519, Japan,

    Kentaro Nakagawa

  3. Department of Biochemistry, Wellcome Trust Centre for Stem Cell Research, Medical Research Council Centre for Stem Cell Biology and Regenerative Medicine, University of Cambridge, Tennis Court Road, Cambridge CB2 1QR, UK,

    Brian Hendrich

Authors
  1. Cristina Aguilera
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  6. Axel Behrens
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Contributions

C.A. designed and performed most of the experiments, analysed data, and co-wrote the paper. K.N. identified Mbd3 as binding specifically to unphosphorylated c-Jun using the yeast three-hybrid screen, generated reagents and provided Fig. 1a, c and Supplementary Fig. 3. R.S. helped with all in vivo experiments and provided Fig. 3a and Supplementary Fig. 7a. A.C. provided Fig. 1f–i. B.H. generated the mbd3 floxed mouse line. A.B. supervised all aspects of this work and wrote the paper.

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Correspondence to Axel Behrens.

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The authors declare no competing financial interests.

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Aguilera, C., Nakagawa, K., Sancho, R. et al. c-Jun N-terminal phosphorylation antagonises recruitment of the Mbd3/NuRD repressor complex. Nature 469, 231–235 (2011). https://doi.org/10.1038/nature09607

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