Binding of 14-3-3β Regulates the Kinase Activity and Subcellular Localization of Testicular Protein Kinase 1

Junko Y. Toshima, Jiro Toshima, Takehiko Watanabe, Kensaku Mizuno

    Research output: Contribution to journalArticle

    35 Citations (Scopus)

    Abstract

    Testicular protein kinase 1 (TESK1) is a serine/threonine kinase that phosphorylates cofilin and induces actin cytoskeletal reorganization. The kinase activity of TESK1 is stimulated by integrin-mediated signaling pathways, but the mechanism of regulation has remained unknown. By using the yeast two-hybrid system, we identified 14-3-3β to be the binding protein of TESK1. Specific interaction between TESK1 and 14-3-3β became evident in in vitro and in vivo co-precipitation assays. 14-3.3β interacts with TESK1 through the C-terminal region of TESK1 and in a manner dependent on the phosphorylation of Ser-439 within an RXXSXP motif. Binding of 14-3-3β inhibited the kinase activity of TESK1. During cell spreading on fibronectin, the TESK1/14-3-3β interaction significantly decreased, in a time course that inversely correlated with increase in TESK1 kinase activity. Thus, the dissociation of 14-3-3β from a TESK1/14-3-3β complex is likely to be involved in the integrin-mediated TESK1 activation. In HeLa cells, TESK1, together with 14-3-3β, accumulated at the cell periphery when cells were plated on fibronectin, whereas they were diffusely distributed in the cytoplasm in the case of non-stimulated cells. We propose that 14-3-3β plays important roles in regulating the kinase activity of TESK1 and localizing TESK1 to cell adhesion sites following integrin stimulation.

    Original languageEnglish
    Pages (from-to)43471-43481
    Number of pages11
    JournalJournal of Biological Chemistry
    Volume276
    Issue number46
    DOIs
    Publication statusPublished - 2001 Nov 16

    Fingerprint

    Phosphotransferases
    Integrins
    testis-specific protein kinase 1
    Fibronectins
    Actin Depolymerizing Factors
    Two-Hybrid System Techniques
    Phosphorylation
    Protein-Serine-Threonine Kinases
    Cell adhesion
    Coprecipitation
    Hybrid systems
    HeLa Cells
    Cell Adhesion
    Yeast
    Actins
    Assays
    Carrier Proteins
    Cytoplasm
    Chemical activation

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Binding of 14-3-3β Regulates the Kinase Activity and Subcellular Localization of Testicular Protein Kinase 1. / Toshima, Junko Y.; Toshima, Jiro; Watanabe, Takehiko; Mizuno, Kensaku.

    In: Journal of Biological Chemistry, Vol. 276, No. 46, 16.11.2001, p. 43471-43481.

    Research output: Contribution to journalArticle

    Toshima, Junko Y. ; Toshima, Jiro ; Watanabe, Takehiko ; Mizuno, Kensaku. / Binding of 14-3-3β Regulates the Kinase Activity and Subcellular Localization of Testicular Protein Kinase 1. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 46. pp. 43471-43481.
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