Regulation of mitogen-activated protein kinases by a calcium/calmodulin-dependent protein kinase cascade

Hervé Enslen, Hiroshi Tokumitsu, Philip Stork, Roger J. Davis, Thomas Soderling

Research output: Contribution to journalArticle

253 Citations (Scopus)

Abstract

Membrane depolarization of NG108 cells gives rapid (2+/calmodulin-dependent protein kinase IV (CaM-KIV), as well as activation of c-Jun N-terminal kinase (JNK). To investigate whether the Ca2+-dependent activation of mitogen-activated protein kinases (ERK, JNK, and p38) might be mediated by the CaM kinase cascade, we have transfected PC12 cells, which lack CaM-KIV, with constitutively active mutants of CaM kinase kinase and/or CaM-KIV (CaM-KKc and CaM-KIVc, respectively). In the absence of depolarization, CaM-KKc transfection had no effect on Elk-dependent transcription of a luciferase reporter gene, whereas CaM-KIVc alone or in combination with CaM-KKc gave 7- to 10-fold and 60- to 80-fold stimulations, respectively, which were blocked by mitogen-activated protein (MAP) kinase phosphatase cotransfection. When epitope-tagged constructs of MAP kinases were co-transfected with CaM-KKc plus CaM-KIVc, the immunoprecipitated MAP kinases were activated 2-fold (ERK-2) and 7-to 10-fold (JNK-1 and p38). The JNK and p38 pathways were further investigated using specific c-Jun or ATF2-dependent transcriptional assays. We found that c-Jun/ATF2-dependent transcriptions were enhanced 7- to 10-fold by CaM-KIVc and 20- to 30-fold by CaM-KKc plus CaM-KIVc. In the case of the Jun-dependent transcription, this effect was not due to direct phosphorylation of c-Jun by activated CaM-KIV, since transcription was blocked by a dominant-negative JNK and by two MAP kinase phosphatases. Mutation of the phosphorylation site (Thr196) in CaM-KIV, which mediates its activation by CaM-KIV kinase, prevented activation of Elk-1, c-Jun, and ATF2 by the CaM kinase cascade. These results establish a new Ca2+-dependent mechanism for regulating MAP kinase pathways and resultant transcription.

Original languageEnglish (US)
Pages (from-to)10803-10808
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number20
StatePublished - Oct 1 1996

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Calcium-Calmodulin-Dependent Protein Kinase Kinase
Calcium-Calmodulin-Dependent Protein Kinase Type 4
Mitogen-Activated Protein Kinases
Phosphotransferases
Mitogen-Activated Protein Kinase Phosphatases
Phosphorylation
MAP Kinase Signaling System
JNK Mitogen-Activated Protein Kinases
PC12 Cells
Luciferases
Reporter Genes
Transfection
Epitopes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Regulation of mitogen-activated protein kinases by a calcium/calmodulin-dependent protein kinase cascade. / Enslen, Hervé; Tokumitsu, Hiroshi; Stork, Philip; Davis, Roger J.; Soderling, Thomas.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 93, No. 20, 01.10.1996, p. 10803-10808.

Research output: Contribution to journalArticle

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