TY - JOUR
T1 - Imaging of heme/hemeproteins in nucleus of the living cells expressing heme-binding nuclear receptors
AU - Itoh, Ryuhei
AU - Fujita, Ken Ichi
AU - Mu, Anfeng
AU - Kim, Dao Hoang Thien
AU - Tai, Tran Tien
AU - Sagami, Ikuko
AU - Taketani, Shigeru
N1 - Funding Information:
This study was supported in part by Grants from the Ministry of Health, Labor and Welfare of Japan , and from the Ministry of Education, Science, Sports, and Culture of Japan . The authors are also grateful to Asako Kawazoe, Koji Fukuda, and Ayaka Furukawa for their technical support.
PY - 2013/7/11
Y1 - 2013/7/11
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84879689629&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84879689629&partnerID=8YFLogxK
U2 - 10.1016/j.febslet.2013.05.036
DO - 10.1016/j.febslet.2013.05.036
M3 - Article
C2 - 23735699
AN - SCOPUS:84879689629
SN - 0014-5793
VL - 587
SP - 2131
EP - 2136
JO - FEBS Letters
JF - FEBS Letters
IS - 14
ER -