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c-Jun N-terminal kinase (JNK) positively regulates NFATc2 transactivation through phosphorylation within the

Inmaculada Ortega-Pérez1, Eva Cano, Felipe Were

  • 1Centro Nacional de Investigaciones Cardiovasculares (CNIC), Ronda de Poniente 5, Tres Cantos, Madrid 28760, Spain.

The Journal of Biological Chemistry
|March 4, 2005
PubMed

Insights

The c-Jun N-terminal kinase (JNK) pathway up-regulates the activity of nuclear factor of activated T cells c2 (NFATc2), a key immune system regulator. This JNK-mediated phosphorylation of NFATc2 differs from other NFAT members, suggesting distinct roles in immune responses.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Signaling

Background:

  • Nuclear factor of activated T cells (NFAT) transcription factors are crucial regulators of immune system function, controlling cytokine gene expression.
  • NFAT activity is primarily regulated by the phosphatase calcineurin, which mediates dephosphorylation for nuclear translocation and transcriptional activity.

Purpose of the Study:

  • To investigate the role of mitogen-activated protein kinase (MAPK) JNK in regulating NFATc2 activity.
  • To identify specific phosphorylation sites and motifs on NFATc2 targeted by JNK.

Main Methods:

  • Mass spectrometry to identify JNK phosphorylation sites on NFATc2.
  • Transfection studies using chimeric constructs and dominant-negative JNK to assess JNK's effect on NFATc2 transactivation.
  • Site-directed mutagenesis to evaluate the importance of identified phosphorylation sites and SP motifs.

Main Results:

  • JNK was found to phosphorylate at least six residues on NFATc2 in vitro, including critical sites Thr(116) and Ser(170).
  • JNK activation significantly stimulates NFATc2-dependent transcription in Jurkat T lymphocytes.
  • In vivo analysis confirmed JNK pathway activation leads to Thr(116) phosphorylation on NFATc2.

Conclusions:

  • The transcriptional activity of NFATc2 is uniquely up-regulated by JNK, differentiating it from other NFAT family members.
  • JNK-mediated phosphorylation plays a specific and differential role in regulating the function of NFAT family members within the immune system.

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