Real-time imaging of NF-AT nucleocytoplasmic shuttling with a photoswitchable fluorescence protein in live cells

Oh Yeun Kwon, Ick Chan Kwon, Hyun Kyu Song, Hyesung Jeon

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: The transcription factor NF-AT plays a key role in the activation of many early immune response genes and is regulated by subcellular localization. NF-AT translocates from the cytoplasm to the nucleus then returns in response to the intracellular calcium level. Methods: We have investigated NF-AT nucleocytoplasmic shuttling in real-time in living cells using NF-ATc1 tagged with the reversibly photoswitchable fluorescence protein, Dronpa. We monitored both nuclear import and export rate of Dronpa-tagged NF-AT in live cells upon stimulation with ionomycin plus calcium (I + Ca2+) or cyclosporin A (CsA). Results: The results show that NF-AT moved into the nucleus within 3-9 min after stimulation and moved back out into the cytoplasm within 15-50 min after CsA addition. In the absence of stimulation, NF-AT stayed in the cytoplasm as in the cells overexpressing GSK-3β, a calcineurin-opposing regulator. General Significance: This semi-quantitative imaging with constant fluorescence provides the basis to detect the real-time effect by several regulators on NF-AT family proteins.

Original languageEnglish
Pages (from-to)1403-1407
Number of pages5
JournalBiochimica et Biophysica Acta - General Subjects
Volume1780
Issue number12
DOIs
Publication statusPublished - 2008 Dec 1

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Keywords

  • Calcineurin
  • Dronpa
  • GSK-3β
  • NF-AT
  • Nucleocytoplasmic shuttling
  • Real-time imaging

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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