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  • Original Article
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Sprouty4 negatively regulates protein kinase C activation by inhibiting phosphatidylinositol 4,5-biphosphate hydrolysis

Oncogene volume 28, pages 1076–1088 (2009)Cite this article

Abstract

Sproutys have been shown to negatively regulate growth factor-induced extracellular signal-regulated kinase (ERK) activation, and suggested to be an anti-oncogene. However, molecular mechanism of the suppression has not yet been clarified completely. Sprouty4 inhibits vascular endothelial growth factor (VEGF)-A-induced ERK activation, but not VEGF-C-induced ERK activation. It has been shown that VEGF-A-mediated ERK activation is strongly dependent on protein kinase C (PKC), whereas that by VEGF-C is dependent on Ras. This suggests that Sprouty4 inhibits the PKC pathway more specifically than the Ras pathway. In this study, we confirmed that Sprouty4 suppressed various signals downstream of PKC, such as phosphorylation of MARCKS and protein kinase D (PKD), as well as PKC-dependent nuclear factor (NF)-κB activation. Furthermore, Sprouty4 suppressed upstream signals of PKC, such as Ca2+ mobilization, phosphatidylinositol 4,5-biphosphate (PIP2) breakdown and inositol 1,4,5-triphosphate (IP3) production in response to VEGF-A. Those effects were dependent on the C-terminal cysteine-rich region, but not on the N-terminal region of Sprouty4, which is critical for the suppression of fibroblast growth factor (FGF)-mediated ERK activation. Sprouty4 overexpression or deletion of the Sprouty4 gene did not affect phospholipase C (PLC) γ-1 activation, which is an enzyme that catalyzes PIP2 hydrolysis. Moreover, Sprouty4 inhibited not only VEGF-A-mediated PIP2 hydrolysis but also inhibited the lysophosphatidic acid (LPA)-induced PIP2 breakdown that is catalyzed by PLCβ/ɛ activated by G-protein coupled receptor (GPCR). Taken together, Sprouty4 has broader suppression activity for various stimuli than previously thought; it may function as an inhibitor for various types of PLC-dependent signaling as well as for ERK activation.

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Abbreviations

GPCR:

G-protein-coupled receptor

IP3:

inositol 1,4,5-triphosphate

LPA:

lysophosphatidic acid

PIP2:

phosphatidyl inositol 4,5-biphosphate

PKC:

protein kinase C

PLC:

phospholipase C

VEGF:

vascular endothelial growth factor

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Acknowledgements

We thank T Yoshioka and M Ohtsu for technical assistance, Dr Sasaki (Akita University), Dr Kawahara, Dr Okuno and Dr Yokomizo (Kyushu University) for reagents and discussions and Y Nishi for manuscript preparation. This study was supported by special Grants-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the Program for Promotion of Fundamental Studies in Health Sciences of the National Institute of Biomedical Innovation (NIBIO), the Takeda Science Foundation, the Kato Memorial Trust for Nambyo Research, the Mitsubishi Pharma Research Foundation, the Nakatomi Foundation, the Naito Foundation, Astellas Foundation for Research on Metabolic Disorders, the Yakult Bioscience Research Foundation and the Princess Takamatsu Cancer Research Fund.

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Authors and Affiliations

  1. Division of Molecular and Cellular Immunology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan

    T Ayada, K Taniguchi, F Okamoto, R Kato, G Takaesu & A Yoshimura

  2. Department of Otorhinolaryngology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    T Ayada & S Komune

  3. Department of Microbiology and Immunology, Keio University School of Medicine, Tokyo, Japan

    A Yoshimura

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Correspondence to A Yoshimura.

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Ayada, T., Taniguchi, K., Okamoto, F. et al. Sprouty4 negatively regulates protein kinase C activation by inhibiting phosphatidylinositol 4,5-biphosphate hydrolysis. Oncogene 28, 1076–1088 (2009). https://doi.org/10.1038/onc.2008.464

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