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cIAP1 regulates TNF-mediated cdc42 activation and filopodia formation

Oncogene volume 33, pages 5534–5545 (2014)Cite this article

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Abstract

Tumour necrosis factor-α (TNF) is a cytokine endowed with multiple functions, depending on the cellular and environmental context. TNF receptor engagement induces the formation of a multimolecular complex including the TNFR-associated factor TRAF2, the receptor-interaction protein kinase RIP1 and the cellular inhibitor of apoptosis cIAP1, the latter being essential for NF-κB activation. Here, we show that cIAP1 also regulates TNF-induced actin cytoskeleton reorganization through a cdc42-dependent, NF-κB-independent pathway. Deletion of cIAP1 prevents TNF-induced filopodia and cdc42 activation. The expression of cIAP1 or its E3-ubiquitin ligase-defective mutant restores the ability of cIAP1−/− MEFs to produce filopodia, whereas a cIAP1 mutant unable to bind TRAF2 does not. Accordingly, the silencing of TRAF2 inhibits TNF-mediated filopodia formation, whereas silencing of RIP1 does not. cIAP1 directly binds cdc42 and promotes its RhoGDIα-mediated stabilization. TNF decreases cIAP1-cdc42 interaction, suggesting that TNF-induced recruitment of cIAP1/TRAF2 to the receptor releases cdc42, which in turn triggers actin remodeling. cIAP1 also regulates cdc42 activation in response to EGF and HRas-V12 expression. A downregulation of cIAP1 altered the cell polarization, the cell adhesion to endothelial cells and cell intercalation, which are cdc42-dependent processes. Finally, we demonstrated that the deletion of cIAP1 regulated the HRas-V12-mediated transformation process, including anchorage-dependent cell growth, tumour growth in a xenograft model and the development of experimental metastasis in the lung.

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Acknowledgements

We thank Dr J Silke, Dr E Lemichez, Dr S Gasman, Dr CL Day, Dr S Ansieu, Dr R Weil and S Monier for kindly providing plasmids and cell lines. We are grateful to Lydie Desoche, Aziza Aznague, Cedric Seignez and Benoit Simon (FEMTO-ST, CLIPP platform) for their technical assistance. We thank A Bouchot and B Gasquet (CellImaP Imagery Facility), A Hammann (Cytometry platform), V Saint-Giorgio (Animal Facility), A Oudot and B Collin (Precilinal imagery platform, Georges-François Leclerc Center) for the use of the imagery, cytometry and animal facilities. We thank P Meier, K Rajalingam, J Bréard, M David and S Ansieu for helpful discussions. This work was supported by grants from the ‘Comité de Côte d’Or of the Ligue Contre le Cancer’ (LD), the ’Association pour la Recherche sur le Cancer’ (ARC to LD), the Association ‘Cent pour sang la Vie’ (LD), the European Union and the ‘Conseil Régional de Bourgogne’, a French Government grant managed by the French National Research Agency under the program ‘Investissements d’Avenir’ with reference ANR-11-LABX-0021’, and fellowships from the ‘Ministère de l’Enseignement Supérieur et de la Recherche’ of France (to AM, JB, JC), ARC (JC) and the ‘Société Française d’Hématologie’ (AM).

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Author notes

  1. A Marivin and J Berthelet: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut National dela Santé et de la Recherche Médicale (Inserm) UMR866, Faculty of Medicine, Dijon, France

    A Marivin, J Berthelet, J Cartier, S Gemble & L Dubrez

  2. Université de Bourgogne; Institut Fédératif de Recherche (IFR) 100,

    A Marivin, J Berthelet, J Cartier, C Paul, S Gemble & L Dubrez

  3. Institut Fédératif de Recherche (IFR) 100, Dijon, France

    A Marivin, J Berthelet, J Cartier, C Paul, S Gemble & L Dubrez

  4. Ecole Pratique des hautes études (EPHE), Dijon, France

    C Paul

  5. Department of Medicine, McGill University, Montreal, Quebec, Canada

    A Morizot & M Saleh

  6. Institut FEMTO-ST, Université de Franche-Comté, CLIPP, Besançon, France

    W Boireau

  7. Inserm UMR749, Institut Fédératif de Recherche (IFR) 54, Villejuif, France

    J Bertoglio

  8. Institut Gustave Roussy, Institut Fédératif de Recherche (IFR) 54, Villejuif, France

    J Bertoglio & E Solary

  9. UMR1009, Institut Fédératif de Recherche (IFR) 54, Villejuif, France

    E Solary

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  1. A Marivin
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Correspondence to L Dubrez.

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Author contributions

AM and JB performed most of the experiments and analysed the data. JB performed the in vivo experiment and analysis. JC performed additional experiments and data analysis. CP and AM contributed to the in vivo analysis. SG and WB performed the biacore experiments and analysis. MS and JB provided valuable materials and expert evaluation. ES provided expert evaluation and corrected the paper and LD conceived and supervised the project, analysed the data and wrote the paper with input from all authors.

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Marivin, A., Berthelet, J., Cartier, J. et al. cIAP1 regulates TNF-mediated cdc42 activation and filopodia formation. Oncogene 33, 5534–5545 (2014). https://doi.org/10.1038/onc.2013.499

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