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Ras–Erk signaling induces phosphorylation of human TLE1 and downregulates its repressor function

Oncogene volume 36, pages 3729–3739 (2017)Cite this article

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Abstract

Signaling mediated by the Ras-extracellular signal-regulated kinase (Erk) pathway often leads to the phosphorylation of transcriptional regulators, thereby modulating their activity and causing concerted changes in gene expression. In Drosophila, the induction of multiple Ras–Erk pathway target genes depends on prior phosphorylation of the general co-repressor Groucho, a modification that downregulates its repressive function. Here, we show that TLE1, one of the four human Groucho orthologs, is similarly phosphorylated in response to Ras–Erk pathway activation, and that this modification attenuates its capacity to repress transcription. Specifically, unphosphorylated TLE1 dominantly suppresses the induction of Ras–Erk pathway target genes in cultured human cells, and the expression of an unphosphorylatable TLE1 derivative causes severe phenotypes in a transgenic Drosophila model system, whereas a phosphomimetic variant of TLE1 exerts only negligible effects. We present data indicating that TLE1 is rapidly excluded from the nucleus following epidermal growth factor receptor pathway activation, an effect that likely accounts for its inability to mediate effective repression under such conditions. Significantly, we find that unphosphorylated TLE1 blocks oncogenic phenotypes induced by mutated H-Ras in human mammary cells, both in vitro and following their implantation in mice. Collectively, our data strongly indicate that phosphorylation of TLE family members and the consequent downregulation of their repressor function is a key conserved step in the transcriptional responses to Ras–Erk signaling, and possibly a critical event in the tumorigenic effects caused by excessive Ras–Erk pathway activity.

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Acknowledgements

We thank members of our laboratory for continued help and encouragement during this project; David Engelberg, Shulamit Katzav, Oded Meyuhas, Joel Yisraeli and, particularly, Gerardo Jiménez for their insightful comments on the manuscript; Yosef Yarden and Stefano Stifani for providing us with DNA constructs and reagents; Konstantin Kogan, Vered Levin-Salomon and David Engelberg for their assistance with the Erk2 in vitro kinase assays and for contributing recombinant activated Erk2; Ahmad Mreisat for technical help; and Amir Orian for hosting TZ in his laboratory for some of her experiments. Work was supported by grants from the Israel Cancer Research Fund (Project Grant), Israel Science Foundation (Center of Excellence 1772/13 and 1552/16), and the Jan M and Eugenia Król Charitable Foundation to ZP; and by the Israel Cancer Association to HN. TK received a Doctoral Fellowship from the Tsipora and Moshe Levin Foundation. DK is a recipient of a Carole Annette Stevens-Walvish Postdoctoral Fellowship in Cancer Research, a Lady Davis Postdoctoral Fellowship and an Israel Cancer Research Fund Booster Fellowship. ZP is an incumbent of the Lady Davis Professorship in Experimental Medicine and Cancer Research.

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Authors and Affiliations

  1. Department of Developmental Biology and Cancer Research, IMRIC, Faculty of Medicine, The Hebrew University, Jerusalem, Israel

    T Zahavi, T Kushnir, R Lange, E Berger, D Kornspan, R Grossman & Z Paroush

  2. Department of Biochemistry and Molecular Biology, IMRIC, Faculty of Medicine, The Hebrew University, Jerusalem, Israel

    A Maimon, S Anzi, E Shaulian & R Karni

  3. Department of Oncology, Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    D Kornspan & H Nechushtan

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  1. T Zahavi
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Correspondence to Z Paroush.

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Zahavi, T., Maimon, A., Kushnir, T. et al. Ras–Erk signaling induces phosphorylation of human TLE1 and downregulates its repressor function. Oncogene 36, 3729–3739 (2017). https://doi.org/10.1038/onc.2016.517

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