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  • Original Paper
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Notch4 intracellular domain binding to Smad3 and inhibition of the TGF-β signaling

Oncogene volume 24, pages 5365–5374 (2005)Cite this article

Abstract

We present evidence that Notch4ICD attenuates TGF-β signaling. Cells expressing the activated form of the Notch4 receptor (ICD4) were resistant to the growth-inhibitory effects of TGF-β. Notch4ICD was found to bind to Smad2, Smad3 and Smad4 but with higher affinity to Smad3. Deletion analysis showed that binding of Smad3 to ICD4 was mediated by its MH2 domain and was not dependent on the presence of the RAM23 region in ICD4. Using two TGF-β/Activin reporter luciferase assays, RT–PCR and Western blot analysis, we demonstrate that ICD4 and ICD4 δRAM23 inhibit Smad-binding element and 3TP luciferase reporter activity and PAI-1 gene expression. MCF-7 human breast cancer cells express Notch4ICD (ICD4) and are resistant to the growth-inhibitory effects of TGF-β. Blockage of Notch4 processing to ICD4 by γ-secretase inhibitor renders MCF-7 cells sensitive to growth inhibition by TGF-β. The interplay between these two signaling pathways may be a significant determinant during mammary tumorigenesis.

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Acknowledgements

We thank Dr Anita B Roberts and Dr Adam B Glick for critical comments about this manuscript. We thank Dr Kohei Miyazono, Dr Tongwen Wang and Dr Ying Zhang for generous gifts of Smad constructs; Dr Jan Kitajewswi for his generous gift of HaHis-Int3; Dr Anthony J Capobianco, University of Pennsylvania for Human Notch1ICD construct and Dr Michael Reiss for providing the phospho-Smad3 antibody. We thank Ahmed Raafat, Tatsuro Ohta, David McCurdy, Xin liu, Vijayachandra Kinnimulki and Jun Zhou for their helpful discussion and technical assistance.

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Authors and Affiliations

  1. The Tumor Growth Factor Section, Mammary Biology and Tumorigenesis Laboratory, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, 20892, MD, USA

    Youping Sun, Caterina Bianco, Nicholas Kenney, Luigi Strizzi, Dina Raafat, Morihisa Hirota, Nadia I Khan, Brenda Jones & David Salomon

  2. Oncogenetics Section, Mammary Biology and Tumorigenesis Laboratory, Center for Cancer Research, National Cancer Institute, National Institute of Health, Bethesda, 20892, MD, USA

    Youping Sun, William Lowther, Katsuaki Kato, Sharon Bargo & Robert Callahan

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  1. Youping Sun
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Correspondence to David Salomon or Robert Callahan.

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Sun, Y., Lowther, W., Kato, K. et al. Notch4 intracellular domain binding to Smad3 and inhibition of the TGF-β signaling. Oncogene 24, 5365–5374 (2005). https://doi.org/10.1038/sj.onc.1208528

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