Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade

Gary A. Wayman, Hiroshi Tokumitsu, Thomas Soderling

    Research output: Contribution to journalArticle

    69 Citations (Scopus)

    Abstract

    The calmodulin-dependent kinase (CaM-K) cascade, a Ca2+-triggered system involving phosphorylation and activation of CaM-KI and CaM-KIV by CaM kinase kinase (CaM-KK), regulates transcription through direct phosphorylation of transcription factors such as cAMP response element- binding protein. We have shown previously that activated CaM-KIV can activate the mitogen-activated protein kinases (Enslen, H., Tokumitsu, H., Stork, P. J. S., Davis, R. J., and Soderling, T. R. (1996) Proc. Natl. Acad. Sci. U.S.A. 93, 10803-10808), and the present paper describes a novel regulatory cross-talk between cAMP kinase (PKA) and CaM-KK. PKA gave rapid phosphorylation in vitro and in cells of recombinant CaM-KK, resulting in 50- 75% inhibition of CaM-KK activity, part of which was due to suppression of CaM-binding by phosphorylation of Ser458 in the CaM-binding domain. However, the Ser458 Ala mutant, or a truncation mutant in which the CaM- binding and autoinhibitory domains were deleted, was still partially suppressed by PKA-mediated phosphorylation. The second inhibitory site was identified as Thr108 by site-specific mutagenesis. Treatments of COS-7, PC12, hippocampal, or Jurkat cells with the PKA activators forskolin or isoproterenol gave 30-90% inhibition of either endogenous or transfected CaM- KK and/or CaM-KIV activities. These results demonstrate that the CaM kinase cascade is negatively regulated in cells by the cAMP/PKA pathway.

    Original languageEnglish (US)
    Pages (from-to)16073-16076
    Number of pages4
    JournalJournal of Biological Chemistry
    Volume272
    Issue number26
    DOIs
    StatePublished - Jun 27 1997

    Fingerprint

    Calcium-Calmodulin-Dependent Protein Kinases
    Phosphotransferases
    Phosphorylation
    Calcium-Calmodulin-Dependent Protein Kinase Kinase
    Cyclic AMP Response Element-Binding Protein
    Jurkat Cells
    PC12 Cells
    Mutagenesis
    Colforsin
    Site-Directed Mutagenesis
    Mitogen-Activated Protein Kinases
    Isoproterenol
    Transcription
    Calmodulin
    Transcription Factors
    Chemical activation

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade. / Wayman, Gary A.; Tokumitsu, Hiroshi; Soderling, Thomas.

    In: Journal of Biological Chemistry, Vol. 272, No. 26, 27.06.1997, p. 16073-16076.

    Research output: Contribution to journalArticle

    Wayman, Gary A. ; Tokumitsu, Hiroshi ; Soderling, Thomas. / Inhibitory cross-talk by cAMP kinase on the calmodulin-dependent protein kinase cascade. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 26. pp. 16073-16076.
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