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Desuppression of TGF-β signaling via nuclear c-Abl-mediated phosphorylation of TIF1γ/TRIM33 at Tyr-524, -610, and -1048

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Abstract

Protein-tyrosine kinases regulate a broad range of intracellular processes occurring primarily just beneath the plasma membrane. With the greatest care to prevent dephosphorylation, we have shown that nuclear tyrosine phosphorylation regulates global chromatin structural states. However, the roles for tyrosine phosphorylation in the nucleus are poorly understood. Here we identify transcriptional intermediary factor 1-γ (TIF1γ/TRIM33/Ectodermin), which suppresses transforming growth factor-β (TGF-β) signaling through the association with Smad2/3 transcription factor, as a new nuclear substrate of c-Abl tyrosine kinase. Replacement of the three tyrosine residues Tyr-524, -610, and -1048 with phenylalanine (3YF) inhibits c-Abl-mediated phosphorylation of TIF1γ and enhances TIF1γ’s association with Smad3. Importantly, knockdown-rescue experiments show that 3YF strengthens TIF1γ’s ability to suppress TGF-β signaling. Intriguingly, activation of c-Abl by epidermal growth factor (EGF) induces desuppression of TGF-β signaling via enhancing the tyrosine phosphorylation level of TIF1γ. TGF-β together with EGF synergistically provokes desuppressive responses of epithelial-to-mesenchymal transition through tyrosine phosphorylation of TIF1γ. These results suggest that nuclear c-Abl-mediated tyrosine phosphorylation of TIF1γ has a desuppressive role in TGF-β–Smad2/3 signaling.

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Acknowledgements

We are grateful to Dr. Eli Canaani (Weizmann Institute of Science), Dr. Hiroyuki Miyoshi (RIKEN BRC, Tsukuba), Dr. Toshiki Tamura (National Institute of Infectious Diseases, Tokyo), and Dr. Masatoshi Tagawa (Chiba Cancer Center Research Institute, Chiba) for their materials. This work was supported in part by grants-in-aid for Scientific Research 15K07922 (to Naoto Y) and 16K08227 (to Noritaka Y), Global COE Program, and Program for Leading Graduate School (LGS) from the MEXT. KA and SK were G-COE Research Assistants. TH and M.M. were LGS Research Assistants. SK, TH, and MM are JSPS Research Fellows.

Author contributions

RY and Naoto Y conceived the study, designed the experiments, and wrote the manuscript. RY, T Tatewaki, Noritaka Y, and TH performed experiments. KA, SK, TK, and T Tomonaga performed phosphoproteomic analysis. RY, T Tatewaki, KA, TH, SK, MM, IM, and Naoto Y analyzed and discussed the data. All authors approved the manuscript.

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Authors and Affiliations

  1. Laboratory of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, 260-8675, Japan

    Ryuzaburo Yuki, Takashi Tatewaki, Noritaka Yamaguchi, Kazumasa Aoyama, Takuya Honda, Sho Kubota, Mariko Morii & Naoto Yamaguchi

  2. Laboratory of Disease Biology and Molecular Medicine, Graduate School of Medicine, Chiba University, Chiba, 260-8670, Japan

    Ryuzaburo Yuki & Ichiro Manabe

  3. Laboratory of Proteome Research, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka, 567-0085, Japan

    Takahisa Kuga & Takeshi Tomonaga

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  1. Ryuzaburo Yuki
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Yuki, R., Tatewaki, T., Yamaguchi, N. et al. Desuppression of TGF-β signaling via nuclear c-Abl-mediated phosphorylation of TIF1γ/TRIM33 at Tyr-524, -610, and -1048. Oncogene 38, 637–655 (2019). https://doi.org/10.1038/s41388-018-0481-z

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