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Identification of a co-activator that links growth factor signalling to c-Jun/AP-1 activation

Nature Cell Biology volume 12, pages 963–972 (2010)Cite this article

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Abstract

The AP-1 transcription factor c-Jun is essential for cellular proliferation in many cell types, but the molecular link between growth factors and c-Jun activation has been enigmatic. In this study we identify a previously uncharacterized RING-domain-containing protein, RACO-1 (RING domain AP-1 co-activator-1), as a c-Jun co-activator that is regulated by growth factor signalling. RACO-1 interacted with c-Jun independently of amino-terminal phosphorylation, and was both necessary and sufficient for c-Jun/AP-1 activation. Growth factor-mediated stimulation of AP-1 was attributable to MEK/ERK-dependent stabilization of RACO-1 protein. Stimulation of the MEK/ERK pathway strongly promoted Lys 63-linked ubiquitylation of RACO-1, which antagonized Lys 48-linked degradative auto-ubiquitylation of the same Lys residues. RACO-1 depletion reduced cellular proliferation and decreased expression of several growth-associated AP-1 target genes, such as cdc2, cyclinD1 and hb-egf. Moreover, transgenic overexpression of RACO-1 augmented intestinal tumour formation triggered by aberrant Wnt signalling and cooperated with oncogenic Ras in colonic hyperproliferation. Thus RACO-1 is a co-activator that links c-Jun to growth factor signalling and is essential for AP-1 function in proliferation.

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Figure 1: RACO-1 binds to c-Jun independently of N-terminal phosphorylation.
Figure 2: RACO-1 is a MEKK1-dependent c-Jun co-activator regulated by MAP kinase signalling pathways.
Figure 3: Growth factor signalling through the MEK pathway stabilizes RACO-1 protein.
Figure 4: MEK signalling promotes the formation of Lys 63-linked ubiquitin chains that antagonize degradative auto-ubiquitylation.
Figure 5: The residues targeted for MEK1-induced ubiquitylation chain elongation are also critical for degradative poly-ubiquitylation.
Figure 6: RACO-1 is an important regulator of cell-cycle progression and proliferation in NIH3T3 fibroblasts.
Figure 7: Transgenic expression of RACO-1 in vivo enhances APCmin+-induced intestinal tumorigenesis and cooperates with oncogenic Ras in colonic hyperproliferation.

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Acknowledgements

We are grateful to N. Ahn, M. Cobb and A. Hock for providing reagents. We thank B. Spencer-Dean, E. Nye and R. Mitter for technical support; the Molecular Biology Core Facility, Paterson Institute for Cancer Research, Manchester, UK for microarray analysis; and H. Walden for critical reading of the manuscript. The London Research Institute is funded by Cancer Research UK.

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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

    Clare C. Davies, Atanu Chakraborty, Filippo Cipriani & Axel Behrens

  2. Department for Molecular Biomedical Research, Vascular Cell Biology Unit, VIB,

    Katharina Haigh & Jody J. Haigh

  3. Ghent, Belgium

    Katharina Haigh & Jody J. Haigh

  4. Department of Biomedical Molecular Biology, Ghent University,

    Katharina Haigh & Jody J. Haigh

  5. Ghent, Belgium

    Katharina Haigh & Jody J. Haigh

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Contributions

C.D and A.C performed all experiments; C.D and A.B designed and analysed all experiments and wrote the manuscript; K.H and J.H generated R26RACO−1 transgenic mice; F.C generated RACO-1 antibodies.

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Correspondence to Axel Behrens.

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Davies, C., Chakraborty, A., Cipriani, F. et al. Identification of a co-activator that links growth factor signalling to c-Jun/AP-1 activation. Nat Cell Biol 12, 963–972 (2010). https://doi.org/10.1038/ncb2098

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