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TGF-β signalling is regulated by Schnurri-2-dependent nuclear translocation of CLIC4 and consequent stabilization of phospho-Smad2 and 3

Nature Cell Biology volume 11, pages 777–784 (2009)Cite this article

Abstract

CLIC4 (chloride intracellular channel 4), a multifunctional protein that traffics between the cytoplasm and nucleus, interacts with Schnurri-2, a transcription factor in the bone morphogenetic protein (BMP) signalling pathway. Here we show that transforming growth factor β (TGF-β) promotes the expression of CLIC4 and Schnurri-2 as well as their association in the cytoplasm and their translocation to the nucleus. In the absence of CLIC4 or Schnurri-2, TGF-β signalling is abrogated. Direct nuclear targeting of CLIC4 enhances TGF-β signalling and removes the requirement for Schnurri-2. Nuclear CLIC4 associates with phospho (p)-Smad2 and p-Smad3, protecting them from dephosphorylation by nuclear phosphatases. An intact TGF-β signalling pathway is essential for CLIC4-mediated growth-arrest. These results newly identify Schnurri-2 and CLIC4 as modifiers of TGF-β signalling through their stabilization of p-Smad2 and 3 in the nucleus.

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Figure 1: TGF-β enhances the expression and association of CLIC4 and Schnurri-2.
Figure 2: CLIC4 and Schnurri-2 enhance TGF-β signalling.
Figure 3: TGF-β induces nuclear translocation of CLIC4, enhancing TGF-β signalling.
Figure 4: Schnurri-2 is essential for TGF-β-dependent nuclear translocation of CLIC4.
Figure 5: Nuclear CLIC4 associates with and stabilizes p-Smad2 and 3.

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References

  1. Suh, K. S. & Yuspa, S. H. Intracellular chloride channels: critical mediators of cell viability and potential targets for cancer therapy. Curr. Pharm. Des. 11, 2753–2764 (2005).

    Article  CAS  Google Scholar 

  2. Littler, D. R. et al. The intracellular chloride ion channel protein CLIC1 undergoes a redox-controlled structural transition. J. Biol. Chem. 279, 9298–9305 (2004).

    Article  CAS  Google Scholar 

  3. Littler, D. R. et al. Crystal structure of the soluble form of the redox-regulated chloride ion channel protein CLIC4. FEBS J. 272, 4996–5007 (2005).

    Article  CAS  Google Scholar 

  4. Shorning, B. Y., Wilson, D. B., Meehan, R. R. & Ashley, R. H. Molecular cloning and developmental expression of two chloride intracellular channel (CLIC) genes in Xenopus laevis. Dev. Genes Evol. 213, 514–518 (2003).

    Article  CAS  Google Scholar 

  5. Fernandez-Salas, E. et al. mtCLIC/CLIC4, an organellular chloride channel protein, is increased by DNA damage and participates in the apoptotic response to p53. Mol. Cell. Biol. 22, 3610–3620 (2002).

    Article  CAS  Google Scholar 

  6. Shiio, Y. et al. Quantitative proteomic analysis of Myc-induced apoptosis: a direct role for Myc induction of the mitochondrial chloride ion channel, mtCLIC/CLIC4. J. Biol. Chem. 281, 2750–2756 (2006).

    Article  CAS  Google Scholar 

  7. Suh, K. S. et al. The organellular chloride channel protein CLIC4/mtCLIC translocates to the nucleus in response to cellular stress and accelerates apoptosis. J. Biol. Chem. 279, 4632–4641 (2004).

    Article  CAS  Google Scholar 

  8. Suh, K. S. et al. CLIC4 mediates and is required for Ca2+-induced keratinocyte differentiation. J. Cell Sci. 120, 2631–2640 (2007).

    Article  CAS  Google Scholar 

  9. Suh, K. S. et al. Reciprocal modifications of CLIC4 in tumor epithelium and stroma mark malignant progression of multiple human cancers. Clin. Cancer Res. 13, 121–131 (2007).

    Article  CAS  Google Scholar 

  10. Ronnov-Jessen, L., Villadsen, R., Edwards, J. C. & Petersen, O. W. Differential expression of a chloride intracellular channel gene, CLIC4, in transforming growth factor-β1-mediated conversion of fibroblasts to myofibroblasts. Am. J. Pathol. 161, 471–480 (2002).

    Article  CAS  Google Scholar 

  11. Dai, H. et al. The zinc finger protein schnurri acts as a Smad partner in mediating the transcriptional response to decapentaplegic. Dev. Biol. 227, 373–387 (2000).

    Article  CAS  Google Scholar 

  12. Jin, W. et al. Schnurri-2 controls BMP-dependent adipogenesis via interaction with Smad proteins. Dev. Cell 10, 461–471 (2006).

    Article  CAS  Google Scholar 

  13. Udagawa, Y. et al. Schnurri interacts with Mad in a Dpp-dependent manner. Genes Cells 5, 359–369 (2000).

    Article  CAS  Google Scholar 

  14. Liu, X. et al. Conditional epidermal expression of TGFβ 1 blocks neonatal lethality but causes a reversible hyperplasia and alopecia. Proc. Natl Acad. Sci. USA 98, 9139–9144 (2001).

    Article  CAS  Google Scholar 

  15. Pietenpol, J. A. et al. TGF-β1 inhibition of c-myc transcription and growth in keratinocytes is abrogated by viral transforming proteins with pRB binding domains. Cell 61, 777–785 (1990).

    Article  CAS  Google Scholar 

  16. Datto, M. B. et al. Transforming growth factor β induces the cyclin-dependent kinase inhibitor p21 through a p53-independent mechanism. Proc. Natl Acad. Sci. USA 92, 5545–5549 (1995).

    Article  CAS  Google Scholar 

  17. Mavrakis, K. J. et al. Arkadia enhances Nodal/TGF-β signaling by coupling phospho-Smad2/3 activity and turnover. PLoS Biol. 5, e67 (2007).

    Article  Google Scholar 

  18. Lin, X. et al. PPM1A functions as a Smad phosphatase to terminate TGFβ signaling. Cell 125, 915–928 (2006).

    Article  CAS  Google Scholar 

  19. Hill, C. S. Nucleocytoplasmic shuttling of Smad proteins. Cell Res. 19, 36–46 (2009).

    Article  CAS  Google Scholar 

  20. Levy, L. et al. Arkadia activates Smad3/Smad4-dependent transcription by triggering signal-induced SnoN degradation. Mol. Cell. Biol. 27, 6068–6083 (2007).

    Article  CAS  Google Scholar 

  21. Varelas, X. et al. TAZ controls Smad nucleocytoplasmic shuttling and regulates human embryonic stem-cell self-renewal. Nature Cell Biol. 10, 837–848 (2008).

    Article  CAS  Google Scholar 

  22. Lichti, U., Anders, J. & Yuspa, S. H. Isolation and short-term culture of primary keratinocytes, hair follicle populations and dermal cells from newborn mice and keratinocytes from adult mice for in vitro analysis and for grafting to immunodeficient mice. Nature Protoc. 3, 799–810 (2008).

    Article  CAS  Google Scholar 

  23. Takagi, T., Harada, J. & Ishii, S. Murine Schnurri-2 is required for positive selection of thymocytes. Nature Immunol. 2, 1048–1053 (2001).

    Article  CAS  Google Scholar 

  24. Fields, S. & Song, O. A novel genetic system to detect protein–protein interactions. Nature 340, 245–246 (1989).

    Article  CAS  Google Scholar 

  25. Fernandez-Salas, E., Sagar, M., Cheng, C., Yuspa, S. H. & Weinberg, W. C. p53 and tumor necrosis factor α regulate the expression of a mitochondrial chloride channel protein. J. Biol. Chem. 274, 36488–36497 (1999).

    Article  CAS  Google Scholar 

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Acknowledgements

The authors thank L. Wakefield for critically reading the manuscript, M. Anzano for help with genotyping, C. Cheng for help with immunoprecipitation assays and colleagues for providing constructive criticism throughout the study. This work was supported by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, NIH.

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  1. Kwang S. Suh

    Present address: Current address: Tumor Bank and Genomics Program, Hackensack University Medical Center, Hackensack, NJ 07601.,

Authors and Affiliations

  1. Laboratory of Cancer Biology and Genetics, 37 Convent Drive, National Cancer Institute, National Institutes of Health, Bethesda, 20892, MD

    Anjali Shukla, Mariam Malik, Christophe Cataisson, Yan Ho, Travis Friesen, Kwang S. Suh & Stuart H. Yuspa

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Contributions

A.S. designed and carried out studies, analysed data and wrote the manuscript; C.C. performed the analysis of in vivo samples and contributed to writing the manuscript; Y.H. and T.F. performed immunoblotting, immunofluorescence and luciferase assays; M.M. designed the yeast two-hybrid study;, K.S.H. provided essential reagents; S.H.Y. designed experiments, analysed and organized the data and wrote the manuscript. All authors proofread the manuscript.

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Correspondence to Stuart H. Yuspa.

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Shukla, A., Malik, M., Cataisson, C. et al. TGF-β signalling is regulated by Schnurri-2-dependent nuclear translocation of CLIC4 and consequent stabilization of phospho-Smad2 and 3. Nat Cell Biol 11, 777–784 (2009). https://doi.org/10.1038/ncb1885

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