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TGF-β-mediated phosphorylation of hnRNP E1 induces EMT via transcript-selective translational induction of Dab2 and ILEI

Nature Cell Biology volume 12, pages 286–293 (2010)Cite this article

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Abstract

Transforming growth factor-β (TGF-β) induces epithelial–mesenchymal transdifferentiation (EMT) accompanied by cellular differentiation and migration1,2,3,4,5. Despite extensive transcriptomic profiling, the identification of TGF-β-inducible, EMT-specific genes has met with limited success. Here we identify a post-transcriptional pathway by which TGF-β modulates the expression of EMT-specific proteins and of EMT itself. We show that heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) binds a structural, 33-nucleotide TGF-β-activated translation (BAT) element in the 3′ untranslated region of disabled-2 (Dab2) and interleukin-like EMT inducer (ILEI) transcripts, and represses their translation. TGF-β activation leads to phosphorylation at Ser 43 of hnRNP E1 by protein kinase Bβ/Akt2, inducing its release from the BAT element and translational activation of Dab2 and ILEI messenger RNAs. Modulation of hnRNP E1 expression or its post-translational modification alters the TGF-β-mediated reversal of translational silencing of the target transcripts and EMT. These results suggest the existence of a TGF-β-inducible post-transcriptional regulon that controls EMT during the development and metastatic progression of tumours.

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Figure 1: TGF-β translationally upregulates Dab2 expression.
Figure 2: The 3′ UTR of Dab2 mRNA contains a cis-regulatory (BAT) element, which is also present in ILEI mRNA.
Figure 3: hnRNP E1 is an integral functional component of the mRNP complex.
Figure 4: Phosphorylation of hnRNP E1 at Ser 43 by TGF-β-mediated activation of Akt2 disrupts its binding to the BAT element and activates the translation of Dab2 and ILEI.
Figure 5: Modulation of hnRNP E1 expression or its post-translational modification alters the translation of Dab2 and ILEI and the sensitivity of NMuMG cells to TGF-β-induced EMT.

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Acknowledgements

We thank Donna M. Driscoll, Barsanjit Mazumder and members of our laboratory for helpful discussions and critical insights. We value the assistance of Michael T. Kinter and Belinda Willard for liquid chromatography–mass spectrometry; of Judith A. Drazba and John Peterson for imaging; and of Michael Budiman for fast performance liquid chromatography. We thank Rakesh Kumar for the gift of the GST–hnRNP E1 construct, Takashi Kobayashi for the gifts of mouse pCMV14-hnRNP E1-Flag and psiRNA-hH1neo-mouse hnRNP E1, and Harold Moses for giving us the EpRas cell line. This work was supported by grants CA55536 and CA80095 from the National Cancer Institute to P.H.H. A.C. is supported by an American Heart Association (Ohio Valley Affiliate) Pre-doctoral Fellowship 075080B.

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Authors and Affiliations

  1. Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, 44195, Ohio, USA

    Arindam Chaudhury, George S. Hussey, Ge Jin & Philip H. Howe

  2. Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, Ohio, 44195, USA

    Partho S Ray & Paul L. Fox

  3. Department of Biological, Geological and Environmental Sciences, Cleveland State University, 2121 Euclid Avenue, Cleveland, 44115, Ohio, USA

    Arindam Chaudhury & George S. Hussey

  4. Department of Biological Sciences, Case Western Reserve University, School of Dental Medicine, 10900 Euclid Avenue, Cleveland, 44106, Ohio, USA

    Ge Jin

  5. Department of Biology, Indian Institute of Science Education and Research, Kolkata, 700106, India

    Partho S Ray

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Contributions

P.H.H. directed the project. G.J. made the initial observation of uncoupled Dab2 mRNA and protein expression levels. G.S.H. performed the experiments in the EpRas cell line. P.S.R. contributed to the polysome profiling and PatSearch analyses. P.L.F. provided critical insights and expertise throughout. G.J., G.S.H. and A.C. made all the reagents. A.C. performed most of the experiments. P.H.H. and A.C. analysed the data and wrote the paper. All authors reviewed the manuscript.

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Correspondence to Philip H. Howe.

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Chaudhury, A., Hussey, G., Ray, P. et al. TGF-β-mediated phosphorylation of hnRNP E1 induces EMT via transcript-selective translational induction of Dab2 and ILEI. Nat Cell Biol 12, 286–293 (2010). https://doi.org/10.1038/ncb2029

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