Characterization of a Ca2+/calmodulin-dependent protein kinase cascade: Molecular cloning and expression of calcium/calmodulin-dependent protein kinase kinase

Hiroshi Tokumitsu, Hervé Enslen, Thomas Soderling

    Research output: Contribution to journalArticle

    192 Citations (Scopus)

    Abstract

    Recent studies have demonstrated that Ca2+/calmodulin-dependent protein kinase IV (CaM-kinase IV) can mediate Ca2+-dependent regulation of gene expression through the phosphorylation of transcriptional activating proteins. We have previously identified and purified a 68-kDa rat brain CaM-kinase kinase that phosphorylates and increases total and Ca2+-independent activities of CaM-kinase IV (Tokumitsu, H., Brickey, D. A., Gold, J., Hidaka, H., Sikela, J., and Soderling, T. R. (1994) J. Biol. Chem. 269, 28640-28647). Using a partial amino acid sequence of the purified brain kinase, a CaM-kinase kinase cDNA was cloned from a rat brain cDNA library. Northern blot analysis showed that CaM-kinase kinase mRNA (3.4 kilobases) was expressed in rat brain, thymus, and spleen. Sequence analyses revealed that the cDNA encoded a 505-amino acid protein, which contained consensus protein kinase motifs and was 30-40% homologous with members of the CaM-kinase family. Expression of the cDNA in COS-7 cells yielded an apparent 68-kDa CaM-binding protein, which catalyzed in vitro activation in the presence of Mg2+/ATP and Ca2+/ CaM of CaM-kinases I and IV but not of CaM-kinase II. Co-expression of CaM-kinase kinase with CaM-kinase IV gave a 14-fold enhancement of cAMP-response elementbinding protein-dependent gene expression compared with CaM-kinase IV alone. These results are consistent with the hypothesis that CaM-kinases I and IV are regulated through a unique signal transduction cascade involving CaM-kinase kinase.

    Original languageEnglish (US)
    Pages (from-to)19320-19324
    Number of pages5
    JournalJournal of Biological Chemistry
    Volume270
    Issue number33
    StatePublished - Aug 18 1995

    Fingerprint

    Calcium-Calmodulin-Dependent Protein Kinase Kinase
    Calcium-Calmodulin-Dependent Protein Kinases
    Cloning
    Molecular Cloning
    Calcium-Calmodulin-Dependent Protein Kinase Type 4
    Phosphotransferases
    Calcium-Calmodulin-Dependent Protein Kinase Type 1
    Brain
    Rats
    Complementary DNA
    Gene expression
    Amino Acids
    Thymus
    Calcium-Calmodulin-Dependent Protein Kinase Type 2
    Signal transduction
    Amino Acid Motifs
    Phosphorylation
    Proteins
    COS Cells
    Gene Expression Regulation

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Characterization of a Ca2+/calmodulin-dependent protein kinase cascade : Molecular cloning and expression of calcium/calmodulin-dependent protein kinase kinase. / Tokumitsu, Hiroshi; Enslen, Hervé; Soderling, Thomas.

    In: Journal of Biological Chemistry, Vol. 270, No. 33, 18.08.1995, p. 19320-19324.

    Research output: Contribution to journalArticle

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