Redox-dependent Regulation of Nuclear Import of the Glucocorticoid Receptor

doi:10.1074/jbc.274.15.10363

Redox-dependent Regulation of Nuclear Import of the Glucocorticoid Receptor*

From the ^‡Second Department of Internal Medicine, Asahikawa Medical College, Nishikagura, Asahikawa 078-8510, the ^¶Department of Genetics and Molecular Biology, Institute for Virus Research, Kyoto University, Shogoin, Kawahara-cho, Kyoto 606-8507, the ^‖Department of Biochemistry, Saitama Cancer Center Research Institute, Ina, Saitama 362-0800, Japan, and the ^**Department of Cell and Molecular Biology, Medical Nobel Institute, Karolinska Institutet, Stockholm S-171 77, Sweden

Abstract

A number of transcription factors including the glucocorticoid receptor (GR) are regulated in a redox-dependent fashion. We have previously reported that the functional activity of the GR is suppressed under oxidative conditions and restored in the presence of reducing reagents. In the present study, we have used a chimeric human GR fused to theAequorea green fluorescent protein and demonstrated that both ligand-dependent and -independent nuclear translocation of the GR is impaired under oxidative conditions in living cells. Substitution of Cys-481 for Ser within NL1 of the human GR resulted in reduction of sensitivity to oxidative treatment, strongly indicating that Cys-481 is one of the target amino acids for redox regulation of the receptor. Taken together, we may conclude that redox-dependent regulation of nuclear translocation of the GR constitutes an important mechanism for modulation of glucocorticoid-dependent signal transduction.

Footnotes

↵* This work was supported in part by grants from the Ministry of Health and Welfare of Japan; the Ministry of Education, Science, Sports, and Culture of Japan; Japan Rheumatism Association; Uehara Memorial Foundation; Akiyama Foundation; Japan Research Foundation for Clinical Pharmacology; and Ito Foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵§ To whom correspondence should be addressed: Second Dept. of Internal Medicine, Asahikawa Medical College, Nishikagura, Asahikawa 078-8510, Japan. Tel.: 81-166-68-2451; Fax: 81-166-68-2459; E-mail:hirotnk{at}asahikawa-med.ac.jp.
Abbreviations:

GR

glucocorticoid receptor

hGR

human GR

hsp90

heat shock protein 90

NLS

nuclear localization signal

T-ag

large T-antigen

GFP

green fluorescent protein

CHO

Chinese hamster ovary

DMEM

Dulbecco’s modified essential medium

DCC

dextran-coated charcoal

FCS

fetal calf serum

PBS

phosphate-buffered saline

GST

glutathioneS-transferase

NAC

N-acetyl-l-cysteine

DBD

DNA binding domain

Mops

4-morpholinepropanesulfonic acid
- Received November 3, 1998.
- Revision received December 28, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.