Imaging of heme/hemeproteins in nucleus of the living cells expressing heme-binding nuclear receptors

Ryuhei Itoh, Ken Ichi Fujita, Anfeng Mu, Dao Hoang Thien Kim, Tran Tien Tai, Ikuko Sagami, Shigeru Taketani

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Several factors involved in the core circadian rhythm are PAS domain proteins, one of which, neuronal PAS2 (NPAS2), contains a heme-binding motif. It is thought that heme controls the transcriptional activity of core circadian factors BMAL1-NPAS2, and that the heme-binding nuclear receptor REV-erbα negatively regulates the expression of BMAL1. To examine the role of heme in the nucleus, we expressed nuclear hemeproteins including the nuclear localization signal-added cytoglobin, NPAS2 and REV-erbα. Then, the living cells expressing these proteins were treated with 2′,7′- dichlorodihydrofluorescin diacetate (DCFH-DA). The fluorescent signal derived from DCFH-DA was observed in the nucleus. When the cells were cultured with hemin, the signal of heme in the nucleus increased. Considering that DCFH-DA reacted with heme, we propose that the use of DCFH-DA could be useful in detection of the heme moiety of hemeprotein in vivo.

Original languageEnglish
Pages (from-to)2131-2136
Number of pages6
JournalFEBS Letters
Volume587
Issue number14
DOIs
Publication statusPublished - 11-07-2013
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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