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Proteolytic regulation of nuclear factor of activated T (NFAT) c2 cells and NFAT activity by caspase-3

Wenfang Wu1, Ravi S Misra, Jennifer Q Russell

  • 1Immunobiology Program, Department of Medicine, The University of Vermont College of Medicine, Burlington, Vermont 05405, USA.

The Journal of Biological Chemistry
|February 4, 2006
PubMed

Insights

Nuclear factor of activated T cells (NFATc1 and NFATc2) are rapidly degraded by caspases in T cells. Inhibiting caspase activity enhances NFAT transcriptional activity, revealing a new post-transcriptional regulation mechanism.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • Nuclear factor of activated T cells (NFAT) transcription factors regulate diverse genes in T cells.
  • NFATc1 and NFATc2 are key NFAT members in peripheral T cells.
  • NFAT regulation involves transcriptional control, but post-transcriptional mechanisms are less understood.

Purpose of the Study:

  • To investigate the post-transcriptional regulation of NFATc1 and NFATc2 in T cells.
  • To identify the role of caspases in NFAT degradation.
  • To explore the impact of caspase activity on NFAT transcriptional function.

Main Methods:

  • Analysis of NFATc1 and NFATc2 degradation in apoptotic T cells.
  • In vitro caspase cleavage assays to identify cleavage sites.
  • Site-directed mutagenesis to abolish caspase cleavage sites in NFATc2.
  • Assessment of NFAT transcriptional activity in wild-type and transgenic T cells.

Main Results:

  • NFATc1 and NFATc2 undergo rapid degradation in apoptotic T cells.
  • NFATc2 is highly sensitive to caspase-3 cleavage at specific N-terminal sites.
  • Mutating caspase cleavage sites in NFATc2 enhances its transcriptional activity.
  • Inhibition of caspase activity or expression of c-FLIP(L) reduces NFATc2 protein levels and activity.

Conclusions:

  • Caspase-mediated cleavage represents a novel post-transcriptional regulatory mechanism for NFATc2.
  • This regulation occurs in both apoptotic and non-apoptotic effector T cells.
  • Modulating caspase activity can significantly impact NFAT-driven gene expression.

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