Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5

Hiroyuki Kobayashi, Taro Saito, Ko Sato, Kotaro Furusawa, Tomohisa Hosokawa, Koji Tsutsumi, Akiko Asada, Shinji Kamada, Toshio Ohshima, Shin Ichi Hisanaga

    Research output: Contribution to journalArticle

    18 Citations (Scopus)

    Abstract

    Cdk5 is a member of the cyclin-dependent kinase (Cdk) family. In contrast to other Cdks that promote cell proliferation, Cdk5 plays a role in regulating various neuronal functions, including neuronal migration, synaptic activity, and neuron death. Cdks responsible for cell proliferation need phosphorylation in the activation loop for activation in addition to binding a regulatory subunit cyclin. Cdk5, however, is activated only by binding to its activator, p35 or p39. Furthermore, in contrast to Cdk1 and Cdk2, which are inhibited by phosphorylation at Tyr-15, the kinase activity of Cdk5 is reported to be stimulated when phosphorylated at Tyr-15 by Src family kinases or receptor-type tyrosine kinases. We investigated the activation mechanism of Cdk5 by phosphorylation at Tyr-15. Unexpectedly, however, it was found that Tyr-15 phosphorylation occurred only on monomeric Cdk5, and the coexpression of activators, p35/p25, p39, or Cyclin I, inhibited the phosphorylation. In neuron cultures, too, the activation of Fyn tyrosine kinase did not increase Tyr-15 phosphorylation of Cdk5. Further, phospho-Cdk5 at Tyr-15 was not detected in the p35-bound Cdk5. In contrast, expression of active Fyn increased p35 in neurons. These results indicate that phosphorylation at Tyr-15 is not an activation mechanism of Cdk5 but, rather, indicate that tyrosine kinases could activate Cdk5 by increasing the protein amount of p35. These results call for reinvestigation of how Cdk5 is regulated downstream of Src family kinases or receptor tyrosine kinases in neurons, which is an important signaling cascade in a variety of neuronal activities.

    Original languageEnglish
    Pages (from-to)19627-19636
    Number of pages10
    JournalJournal of Biological Chemistry
    Volume289
    Issue number28
    DOIs
    Publication statusPublished - 2014 Jul 11

    Fingerprint

    Cyclin-Dependent Kinase 5
    Phosphorylation
    Chemical activation
    Neurons
    Protein-Tyrosine Kinases
    src-Family Kinases
    Cell proliferation
    Cyclin I
    Proto-Oncogene Proteins c-fyn
    Cell Proliferation
    Cyclins
    Cyclin-Dependent Kinases
    Receptor Protein-Tyrosine Kinases

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Molecular Biology

    Cite this

    Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5. / Kobayashi, Hiroyuki; Saito, Taro; Sato, Ko; Furusawa, Kotaro; Hosokawa, Tomohisa; Tsutsumi, Koji; Asada, Akiko; Kamada, Shinji; Ohshima, Toshio; Hisanaga, Shin Ichi.

    In: Journal of Biological Chemistry, Vol. 289, No. 28, 11.07.2014, p. 19627-19636.

    Research output: Contribution to journalArticle

    Kobayashi, H, Saito, T, Sato, K, Furusawa, K, Hosokawa, T, Tsutsumi, K, Asada, A, Kamada, S, Ohshima, T & Hisanaga, SI 2014, 'Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5', Journal of Biological Chemistry, vol. 289, no. 28, pp. 19627-19636. https://doi.org/10.1074/jbc.M113.501148
    Kobayashi, Hiroyuki ; Saito, Taro ; Sato, Ko ; Furusawa, Kotaro ; Hosokawa, Tomohisa ; Tsutsumi, Koji ; Asada, Akiko ; Kamada, Shinji ; Ohshima, Toshio ; Hisanaga, Shin Ichi. / Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5. In: Journal of Biological Chemistry. 2014 ; Vol. 289, No. 28. pp. 19627-19636.
    @article{9cc9a6b38119405d8602874715e912c6,
    title = "Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5",
    abstract = "Cdk5 is a member of the cyclin-dependent kinase (Cdk) family. In contrast to other Cdks that promote cell proliferation, Cdk5 plays a role in regulating various neuronal functions, including neuronal migration, synaptic activity, and neuron death. Cdks responsible for cell proliferation need phosphorylation in the activation loop for activation in addition to binding a regulatory subunit cyclin. Cdk5, however, is activated only by binding to its activator, p35 or p39. Furthermore, in contrast to Cdk1 and Cdk2, which are inhibited by phosphorylation at Tyr-15, the kinase activity of Cdk5 is reported to be stimulated when phosphorylated at Tyr-15 by Src family kinases or receptor-type tyrosine kinases. We investigated the activation mechanism of Cdk5 by phosphorylation at Tyr-15. Unexpectedly, however, it was found that Tyr-15 phosphorylation occurred only on monomeric Cdk5, and the coexpression of activators, p35/p25, p39, or Cyclin I, inhibited the phosphorylation. In neuron cultures, too, the activation of Fyn tyrosine kinase did not increase Tyr-15 phosphorylation of Cdk5. Further, phospho-Cdk5 at Tyr-15 was not detected in the p35-bound Cdk5. In contrast, expression of active Fyn increased p35 in neurons. These results indicate that phosphorylation at Tyr-15 is not an activation mechanism of Cdk5 but, rather, indicate that tyrosine kinases could activate Cdk5 by increasing the protein amount of p35. These results call for reinvestigation of how Cdk5 is regulated downstream of Src family kinases or receptor tyrosine kinases in neurons, which is an important signaling cascade in a variety of neuronal activities.",
    author = "Hiroyuki Kobayashi and Taro Saito and Ko Sato and Kotaro Furusawa and Tomohisa Hosokawa and Koji Tsutsumi and Akiko Asada and Shinji Kamada and Toshio Ohshima and Hisanaga, {Shin Ichi}",
    year = "2014",
    month = "7",
    day = "11",
    doi = "10.1074/jbc.M113.501148",
    language = "English",
    volume = "289",
    pages = "19627--19636",
    journal = "Journal of Biological Chemistry",
    issn = "0021-9258",
    publisher = "American Society for Biochemistry and Molecular Biology Inc.",
    number = "28",

    }

    TY - JOUR

    T1 - Phosphorylation of Cyclin-dependent kinase 5 (Cdk5) at Tyr-15 is inhibited by Cdk5 activators and does not contribute to the activation of Cdk5

    AU - Kobayashi, Hiroyuki

    AU - Saito, Taro

    AU - Sato, Ko

    AU - Furusawa, Kotaro

    AU - Hosokawa, Tomohisa

    AU - Tsutsumi, Koji

    AU - Asada, Akiko

    AU - Kamada, Shinji

    AU - Ohshima, Toshio

    AU - Hisanaga, Shin Ichi

    PY - 2014/7/11

    Y1 - 2014/7/11

    N2 - Cdk5 is a member of the cyclin-dependent kinase (Cdk) family. In contrast to other Cdks that promote cell proliferation, Cdk5 plays a role in regulating various neuronal functions, including neuronal migration, synaptic activity, and neuron death. Cdks responsible for cell proliferation need phosphorylation in the activation loop for activation in addition to binding a regulatory subunit cyclin. Cdk5, however, is activated only by binding to its activator, p35 or p39. Furthermore, in contrast to Cdk1 and Cdk2, which are inhibited by phosphorylation at Tyr-15, the kinase activity of Cdk5 is reported to be stimulated when phosphorylated at Tyr-15 by Src family kinases or receptor-type tyrosine kinases. We investigated the activation mechanism of Cdk5 by phosphorylation at Tyr-15. Unexpectedly, however, it was found that Tyr-15 phosphorylation occurred only on monomeric Cdk5, and the coexpression of activators, p35/p25, p39, or Cyclin I, inhibited the phosphorylation. In neuron cultures, too, the activation of Fyn tyrosine kinase did not increase Tyr-15 phosphorylation of Cdk5. Further, phospho-Cdk5 at Tyr-15 was not detected in the p35-bound Cdk5. In contrast, expression of active Fyn increased p35 in neurons. These results indicate that phosphorylation at Tyr-15 is not an activation mechanism of Cdk5 but, rather, indicate that tyrosine kinases could activate Cdk5 by increasing the protein amount of p35. These results call for reinvestigation of how Cdk5 is regulated downstream of Src family kinases or receptor tyrosine kinases in neurons, which is an important signaling cascade in a variety of neuronal activities.

    AB - Cdk5 is a member of the cyclin-dependent kinase (Cdk) family. In contrast to other Cdks that promote cell proliferation, Cdk5 plays a role in regulating various neuronal functions, including neuronal migration, synaptic activity, and neuron death. Cdks responsible for cell proliferation need phosphorylation in the activation loop for activation in addition to binding a regulatory subunit cyclin. Cdk5, however, is activated only by binding to its activator, p35 or p39. Furthermore, in contrast to Cdk1 and Cdk2, which are inhibited by phosphorylation at Tyr-15, the kinase activity of Cdk5 is reported to be stimulated when phosphorylated at Tyr-15 by Src family kinases or receptor-type tyrosine kinases. We investigated the activation mechanism of Cdk5 by phosphorylation at Tyr-15. Unexpectedly, however, it was found that Tyr-15 phosphorylation occurred only on monomeric Cdk5, and the coexpression of activators, p35/p25, p39, or Cyclin I, inhibited the phosphorylation. In neuron cultures, too, the activation of Fyn tyrosine kinase did not increase Tyr-15 phosphorylation of Cdk5. Further, phospho-Cdk5 at Tyr-15 was not detected in the p35-bound Cdk5. In contrast, expression of active Fyn increased p35 in neurons. These results indicate that phosphorylation at Tyr-15 is not an activation mechanism of Cdk5 but, rather, indicate that tyrosine kinases could activate Cdk5 by increasing the protein amount of p35. These results call for reinvestigation of how Cdk5 is regulated downstream of Src family kinases or receptor tyrosine kinases in neurons, which is an important signaling cascade in a variety of neuronal activities.

    UR - http://www.scopus.com/inward/record.url?scp=84904182164&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84904182164&partnerID=8YFLogxK

    U2 - 10.1074/jbc.M113.501148

    DO - 10.1074/jbc.M113.501148

    M3 - Article

    C2 - 24872417

    AN - SCOPUS:84904182164

    VL - 289

    SP - 19627

    EP - 19636

    JO - Journal of Biological Chemistry

    JF - Journal of Biological Chemistry

    SN - 0021-9258

    IS - 28

    ER -