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PKCδ-mediated phosphorylation of BAG3 at Ser187 site induces epithelial−mesenchymal transition and enhances invasiveness in thyroid cancer FRO cells

Oncogene volume 32, pages 4539–4548 (2013)Cite this article

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Abstract

Protein kinase C delta (PKCδ) is a serine (Ser)/threonine kinase, which regulates numerous cellular processes, including proliferation, differentiation, migration and apoptosis. In the current study, Chinese hamster ovary cells were transfected with either a constitutively activated PKCδ or a dominant negative PKCδ, phosphoprotein enrichment, two-dimensional difference gel electrophoresis and mass spectrometry was combined to globally identified candidates of PKCδ cascade. We found that Bcl-2 associated athanogene 3 (BAG3) was one of the targets of PKCδ cascade, and BAG3 interacted with PKCδ in vivo. In addition, we clarified that BAG3 was phosphorylate at Ser187 site in a PKCδ-dependent manner in vivo. BAG3 has been implicated in multiple cellular functions, including proliferation, differentiation, apoptosis, migration, invasion, macroautophagy and so on. We generated wild-type (WT)-, Ser187Ala (S187A)- or Ser187Asp (S187D)-BAG3 stably expressing FRO cells, and noticed that phosphorylation state of BAG3 influenced FRO morphology. Finally, for the first time, we showed that BAG3 was implicated in epithelial−mesenchymal transition (EMT) procedure, and phosphorylation state at Ser187 site had a critical role in EMT regulation by BAG3. Collectively, the current study indicates that BAG3 is a novel substrate of PKCδ, and PKCδ-mediated phosphorylation of BAG3 is implicated in EMT and invasiveness of thyroid cancer cells.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (31070697 and 31170727), Program for LNET (LJQ2011083) and Foundation of Liaoning Educational Committee (L2010561) to H-Q Wang.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, China Medical University, Shenyang, China

    N Li, Z-H Zong, B-Q Liu, C Li, Q Zhang & H-Q Wang

  2. Key Laboratory of Cell Biology, Ministry of Public Health and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang, China

    N Li & H-Q Wang

  3. Department of Endocrinology & Metabolism, the First Affiliated Hospital, China Medical University, Shenyang, China

    Z-X Du

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Correspondence to H-Q Wang.

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Li, N., Du, ZX., Zong, ZH. et al. PKCδ-mediated phosphorylation of BAG3 at Ser187 site induces epithelial−mesenchymal transition and enhances invasiveness in thyroid cancer FRO cells. Oncogene 32, 4539–4548 (2013). https://doi.org/10.1038/onc.2012.466

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