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Splicing switch of an epigenetic regulator by RNA helicases promotes tumor-cell invasiveness

Nature Structural & Molecular Biology volume 19, pages 1139–1146 (2012)Cite this article

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Abstract

Both epigenetic and splicing regulation contribute to tumor progression, but the potential links between these two levels of gene-expression regulation in pathogenesis are not well understood. Here, we report that the mouse and human RNA helicases Ddx17 and Ddx5 contribute to tumor-cell invasiveness by regulating alternative splicing of several DNA- and chromatin-binding factors, including the macroH2A1 histone. We show that macroH2A1 splicing isoforms differentially regulate the transcription of a set of genes involved in redox metabolism. In particular, the SOD3 gene that encodes the extracellular superoxide dismutase and plays a part in cell migration is regulated in an opposite manner by macroH2A1 splicing isoforms. These findings reveal a new regulatory pathway in which splicing factors control the expression of histone variant isoforms that in turn drive a transcription program to switch tumor cells to an invasive phenotype.

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Figure 1: The Ddx5 and Ddx17 RNA helicases contribute to cell migration and invasion.
Figure 2: The Ddx5 and Ddx17 RNA helicases control alternative splicing of DNA- and chromatin-binding factors.
Figure 3: Alternative splicing of the macroH2A1 histone variant is involved in tumor progression.
Figure 4: Alternative splicing of the macroH2A1 histone variant contributes to the Ddx5-Ddx17 effect on cell migration and invasion.
Figure 5: Regulation of mH2A1 splicing isoforms by Ddx5 and Ddx17 affects regulation of genes involved in redox metabolism.
Figure 6: Ddx5 and Ddx17–regulated SOD3 expression is involved in cell migration and invasion.
Figure 7: mH2A1 associates with the SOD3 gene, and its splicing variants regulate SOD3 transcription activity in an opposite manner.
Figure 8: mH2A1 alternative splicing controlled by Ddx5 and Ddx17 in MDA-MB-231 cells.

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Acknowledgements

We thank M. Buschbeck and V.A. Raker for critically reading the manuscript and L. Mhamdi for technical assistance. We thank C. Vanbelle and C. Boucharon from CeCILE - SFR Sante Lyon-Est for technical assistance and helpful discussion. This work was supported by the Institut National du Cancer (2008-1-PL BIO 01 to D.A. and S.V.), Agence Nationale de la Recherche (BLAN07-3_186592 to D.A. and S.V.), Association d'Aide à la Recherche Cancérologique de Saint-Cloud (to R.L.) and Fondation Recherche Médicale (Equipe FRM DEQ20110421278). S.G. was supported by Association pour la Recherche sur le Cancer; L.G. and S.P. by Agence Nationale de la Recherche; E.Z., H.M. and J.-P.V. by FRM; E.D. by Ligue Nationale Contre le Cancer; and M.P.E. by Association Française contre les Myopathies.

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  1. Etienne Dardenne and Sandra Pierredon: These authors contributed equally to this work.

Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale, U1052, Centre de Recherche en Cancérologie de Lyon, Lyon, France.,

    Etienne Dardenne, Lise Gratadou, Micaela Polay Espinoza, Eleonora Zonta, Sophie Germann, Hussein Mortada, Jean-Philippe Villemin, Martin Dutertre & Didier Auboeuf

  2. Institut National de la Santé et de la Recherche Médicale, U563, Institut Claudius Regaud, Toulouse, France

    Sandra Pierredon, Magali Lacroix-Triki & Stéphan Vagner

  3. Institut National de la Santé et de la Recherche Médicale, U981, Institut de Cancérologie Gustave Roussy, Villejuif, France

    Sandra Pierredon, Magali Lacroix-Triki & Stéphan Vagner

  4. Laboratory of Oncogenetics, Institut Curie–Hôpital René Huguenin, St-Cloud, France

    Keltouma Driouch & Rosette Lidereau

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Contributions

E.D., S.P., M.D., R.L., S.V. and D.A. designed the experiments; S.V. and D.A. supervised the project; E.D., S.P., K.D., L.G., M.L.-T., E.Z. and S.G. conducted the experiments; M.P.E., H.M. and J.-P.V. performed all bioinformatics analysis. All authors performed the data analysis and wrote the manuscript.

Corresponding authors

Correspondence to Stéphan Vagner or Didier Auboeuf.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 and Supplementary Table 5 (PDF 1113 kb)

Supplementary Table 1

Genes affected at the transcriptional level by ddx5 and ddx17 depletion. (XLS 253 kb)

Supplementary Table 2

Genes affected at the exon level by ddx5 and ddx17 depletion. (XLS 1373 kb)

Supplementary Table 3

Genes affected at the transcriptional level by mH2A1.1 and mH2A1.2 depletion. (XLS 156 kb)

Supplementary Table 4

Sequences of siRNAs and primers (XLS 38 kb)

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Dardenne, E., Pierredon, S., Driouch, K. et al. Splicing switch of an epigenetic regulator by RNA helicases promotes tumor-cell invasiveness. Nat Struct Mol Biol 19, 1139–1146 (2012). https://doi.org/10.1038/nsmb.2390

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