Roles of Calmodulin-dependent Protein Kinases and Phosphatase in Calcium-dependent Transcription of Immediate Early Genes

Hervé Enslen, Thomas Soderling

    Research output: Contribution to journalArticle

    146 Citations (Scopus)

    Abstract

    Recent studies indicate multiple mechanisms are involved in Ca2+ stimulation of gene expression. We have used cell-permeable, specific inhibitors of calmodulin-dependent protein kinases (CaM kinases) and phosphatase (calcineurin) to investigate the involvement of these enzymes in transcriptional regulation of three immediate early genes in PC12 cells stimulated with A23187 or KCl. Preincubation of PC12 cells with the CaM kinase inhibitor KN-62 blocked autophosphorylation of CaM kinase II in response to stimulation by the Ca2+ ionophore A23187. KN-62 treatment also resulted in a 60-70% inhibition of Ca2+-dependent transcription of c-fos, NGFI-A (zif 268), and NGFI-B (nur 77) as assessed by either Northern or nuclear run-on analyses. Preincubation with the calcineurin inhibitors FK-506 or cyclosporin A strongly enhanced expression of NGFI-A and blocked transcription of NGFI-B, but it had no significant effect on Ca2+-stimulated transcription of c-fos. Both FK-506 and KN-62 were specific for Ca2+-stimulated transcription as neither effected transcription in response to forskolin or phorbol ester (12-O-tetradecanoylphorbol-13-acetate) treatment. This is the first report of CaM kinase and calcineurin involvement in transcriptional regulation of NGFI-A and NGFI-B. Activation of CaM kinases and calcineurin, in response to elevated intracellular Ca2+, would exert antagonistic effects on transcription of NGFI-A- Since inhibition of either the kinase or phosphatase decreased transcription of NGFI-B by 60-90%, this suggests that each enzyme is necessary but not sufficient for Ca2+ stimulation. These results indicate that CaM kinases and calcineurin can mediate broad and complex regulation of Ca2+-stimulated gene expression.

    Original languageEnglish (US)
    Pages (from-to)20872-20877
    Number of pages6
    JournalJournal of Biological Chemistry
    Volume269
    Issue number33
    StatePublished - Aug 19 1994

    Fingerprint

    KN 62
    Calcium-Calmodulin-Dependent Protein Kinases
    Immediate-Early Genes
    Phosphoprotein Phosphatases
    Transcription
    Calcineurin
    Genes
    Calcium
    PC12 Cells
    Calcimycin
    Tacrolimus
    Gene expression
    Gene Expression
    Calcium-Calmodulin-Dependent Protein Kinase Type 2
    Ionophores
    Phorbol Esters
    Tetradecanoylphorbol Acetate
    Colforsin
    Enzymes
    Protein Kinase Inhibitors

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Roles of Calmodulin-dependent Protein Kinases and Phosphatase in Calcium-dependent Transcription of Immediate Early Genes. / Enslen, Hervé; Soderling, Thomas.

    In: Journal of Biological Chemistry, Vol. 269, No. 33, 19.08.1994, p. 20872-20877.

    Research output: Contribution to journalArticle

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