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NFAT indicates nucleocytoplasmic damped oscillation via its feedback modulator.

Masashi Muramatsu1, Takeshi Ito2, Hokuto Shimoji1

  • 1Divison of Molecular and Vascular Biology, Institute of Resource Development and Analysis, Kumamoto University, Kumamoto, Japan.

Biochemical and Biophysical Research Communications
|July 31, 2021
PubMed
Summary

Down syndrome critical region (DSCR)-1 protein regulates Nuclear Factor of Activated T-cells 1 (NFAT1) oscillations in endothelial cells. DSCR-1 reduction causes NFAT1 nuclear retention, impacting cell signaling.

Keywords:
CalcineurinDSCR-1EndotheliumMathematical analysisNFATOscillation

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Systems Biology

Background:

  • Cell signaling and gene regulation maintain homeostasis.
  • Nuclear Factor-kappa B (NF-κB) and Inhibitor of kappa B (IκB) form a feedback loop.
  • Nuclear Factor of Activated T-cells (NFAT) signaling is crucial in cellular responses.

Purpose of the Study:

  • To investigate the regulatory role of Down syndrome critical region (DSCR)-1 in NFAT1 nucleocytoplasmic transport.
  • To elucidate the molecular mechanism behind NFAT1 oscillation in endothelial cells under stimulation.
  • To develop and validate a mathematical model for NFAT1 spatiotemporal dynamics.

Main Methods:

  • Primary endothelial cell culture.
  • Stimulation with shear stress and Vascular Endothelial Growth Factor (VEGF).
  • NFAT1 and DSCR-1 expression analysis.
  • Mathematical modeling of NFAT1, calcineurin, and DSCR-1 interactions.
  • DSCR-1 knockdown experiments.

Main Results:

  • Shear stress and VEGF treatment induced rapid NFAT1 nuclear localization and DSCR-1 transactivation.
  • DSCR-1 transactivation led to NFAT1 cytoplasmic sequestration, causing damped NFAT1 oscillations.
  • Mathematical model predicted that DSCR-1 reduction would result in nuclear NFAT1 retention and loss of oscillation.
  • DSCR-1 knockdown confirmed this prediction, increasing dephosphorylated NFAT1 in the nucleus and cytoplasm, leading to nuclear retention.

Conclusions:

  • DSCR-1 is a critical regulator of NFAT1 nucleocytoplasmic oscillation in endothelial cells.
  • Sustained shear stress or VEGF stimulation leads to NFAT1 oscillation modulated by DSCR-1.
  • The developed mathematical model accurately reproduces experimental NFAT1 dynamics.
  • Combined experimental and mathematical approaches offer quantitative insights into NFAT1 feedback systems.