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Nuclear accumulation of NFAT4 opposed by the JNK signal transduction pathway
C W Chow1, M Rincón, J Cavanagh
1Howard Hughes Medical Institute, Program in Molecular Medicine, Department of Biochemistry and Molecular Biology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
Abstract:
The nuclear factor of activated T cells (NFAT) group of transcription factors is retained in the cytoplasm of quiescent cells. NFAT activation is mediated in part by induced nuclear import. This process requires calcium-dependent dephosphorylation of NFAT caused by the phosphatase calcineurin. The c-Jun amino-terminal kinase (JNK) phosphorylates NFAT4 on two sites. Mutational removal of the JNK phosphorylation sites caused constitutive nuclear localization of NFAT4. In contrast, JNK activation in calcineurin-stimulated cells caused nuclear exclusion of NFAT4. These findings show that the nuclear accumulation of NFAT4 promoted by calcineurin is opposed by the JNK signal transduction pathway.
Insights
Nuclear import of nuclear factor of activated T cells (NFAT) is key for activation. The JNK pathway opposes calcineurin-mediated nuclear accumulation of NFAT4, revealing a critical regulatory mechanism.
Area of Science:
- Immunology
- Molecular Biology
- Cell Signaling
Background:
- Nuclear factor of activated T cells (NFAT) proteins reside in the cytoplasm of inactive cells.
- NFAT activation involves translocation to the nucleus, a process regulated by calcium signaling and calcineurin phosphatase activity.
Purpose of the Study:
- To investigate the role of c-Jun amino-terminal kinase (JNK) in regulating NFAT4 nuclear localization.
- To elucidate the interplay between calcineurin and JNK signaling pathways in controlling NFAT4 cellular localization.
Main Methods:
- Site-directed mutagenesis to alter JNK phosphorylation sites on NFAT4.
- Analysis of NFAT4 localization in response to calcineurin stimulation and JNK activation.
Main Results:
- Mutating JNK phosphorylation sites on NFAT4 resulted in its constitutive nuclear localization.
- JNK activation in calcineurin-stimulated cells led to NFAT4 nuclear exclusion.
- Demonstrated opposing effects of calcineurin and JNK on NFAT4 nuclear import.
Conclusions:
- The JNK signaling pathway acts antagonistically to the calcineurin pathway in regulating NFAT4 nuclear accumulation.
- JNK phosphorylation is a critical determinant of NFAT4 localization and, consequently, its transcriptional activity.