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Crk-associated substrate lymphocyte type regulates transforming growth factor-β signaling by inhibiting Smad6 and Smad7

Oncogene volume 26, pages 893–904 (2007)Cite this article

Abstract

Crk-associated substrate lymphocyte type (Cas-L) is a 105 kDa docking protein with diverse functional properties, including regulation of cell division, proliferation, migration and adhesion. Cas-L is also involved in β1 integrin- or antigen receptor-mediated signaling in B and T cells. In the present study, we demonstrate that Cas-L potentiates transforming growth factor-β (TGF-β) signaling pathway by interacting with Smad6 and Smad7. Immunoprecipitation experiments reveal that single domain deletion of full-length Cas-L completely abolishes its docking function with Smad6 and Smad7, suggesting that the natural structure of Cas-L is necessary for its association with Smad6 and Smad7. On the other hand, both N-terminal and C-terminal deletion mutants of Smad6 and Smad7 still retain their docking ability to Cas-L, suggesting that Smad6 and Smad7 possess several binding motifs to Cas-L. Moreover, Cas-L interaction with Mad-homology (MH)2 domain, but not with MH1 domain of Smad6 or Smad7, ameliorates TGF-β-induced signaling pathway. Finally, depletion of Cas-L by small-interfering RNA oligo attenuates TGF-β-induced growth inhibition of Huh-7 cells, with a concomitant reduction in phosphorylation of Smad2 and Smad3. These results strongly suggest that Cas-L is a potential regulator of TGF-β signaling pathway.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

Cas-L:

Crk-associated substrate lymphocyte type

TGF-β:

transforming growth factor β

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Acknowledgements

This work was supported by Grant-in-Aid of Ministry of Education, Science, Sports and Culture (CM), and Ministry of Health, Labor, and Welfare, Japan (CM). T I is the recipients of a grant from Osaka Kidney Foundation (OKF05-0002), and The Yasuda Medical Foundation. We are thankful to Dr Takeshi Imamura (Department of Biochemistry, The JFCR Cancer Institute, Tokyo, Japan), and Dr Kohei Miyazono (Department of Molecular Pathology, Graduate School of Medicine, University of Tokyo, Japan) for providing us with constructs essential for the present work. We also thank Mr Hiroyuki Kayo for his technical assistance and advices.

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Author notes

  1. S Inamoto, S Iwata and T Inamoto: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Clinical Immunology, Advanced Clinical Research Center, Institute of Medical Science, The University of Tokyo, Tokyo, Japan

    S Inamoto, S Iwata, T Inamoto, S Nomura, T Sasaki, Y Urasaki, O Hosono, H Kawasaki, H Tanaka & C Morimoto

  2. Department of Medicine, Osaka Medical College, Osaka, Japan

    S Inamoto & T Inamoto

  3. Department of Hematologic Malignancies, Nevada Cancer Institute, Las Vegas, NV, USA

    N H Dang & C Morimoto

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  1. S Inamoto
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Correspondence to C Morimoto.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Inamoto, S., Iwata, S., Inamoto, T. et al. Crk-associated substrate lymphocyte type regulates transforming growth factor-β signaling by inhibiting Smad6 and Smad7. Oncogene 26, 893–904 (2007). https://doi.org/10.1038/sj.onc.1209848

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