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Interdependent action of RalGEF and Erk in Ras-induced primitive endoderm differentiation of F9 embryonal carcinoma cells

Oncogene volume 18, pages 4435–4439 (1999)Cite this article

Abstract

Previous work by us and others has implicated a role for Ral guanine exchange factors (RalGEFs) in Ras-induced cell growth and oncogenic transformation. Here we show for the first time that RalGEFs are involved in Ras-induced differentiation as well. Expression of oncogenic Ras in F9 embryonal carcinoma (EC) cells is known to induce differentiation to a primitive endoderm (PrE)-like phenotype, but the downstream signal transduction mechanisms involved are unclear. We found that PrE differentiation is induced by the Ras effector domain mutants, RasV12G37 and RasV12E38, but not by RasV12C40. Accordingly, expression of constitutively active forms of RalGEF (Rlf-CAAX) or Raf1 (Raf-CAAX) is sufficient to induce differentiation. Inhibition of RalGEF activity by expression of dominant negative Ral completely abolishes Rlf-CAAX- and RasV12G37-induced differentiation, while it reduces differentiation by RasV12 and Raf-CAAX. Finally, while Rlf-CAAX does not increase Erk activity, inhibition of MEK blocks both Ras- as well as Rlf-CAAX-induced differentiation, suggesting that RalGEFs induce PrE differentiation in a manner depending on basal MEK or Erk activity. Based on these results we conclude that Ras induces PrE differentiation of F9 EC cells via an interplay of Erk- and RalGEF-mediated pathways.

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Acknowledgements

We would like to thank Drs J Downward, C Marshall and J Pouysségur for various constructs. This work was supported by Netherlands Organisation for Scientific Research Grant 805-33-202.

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Authors and Affiliations

  1. Laboratory for Physiological Chemistry and Centre for Biomedical Genetics, Utrecht University, Universiteitsweg 100, The Netherlands

    Mark HG Verheijen, Rob MF Wolthuis & Johannes L Bos

  2. Hubrecht Laboratory, the Netherlands Institute for Developmental Biology, Upssalalaan 8, Utrecht, 3584 CT, The Netherlands

    Mark HG Verheijen, Libert HK Defize & Jeroen den Hertog

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  1. Mark HG Verheijen
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Verheijen, M., Wolthuis, R., Defize, L. et al. Interdependent action of RalGEF and Erk in Ras-induced primitive endoderm differentiation of F9 embryonal carcinoma cells. Oncogene 18, 4435–4439 (1999). https://doi.org/10.1038/sj.onc.1202834

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