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PMID: 9891038 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Discrimination between NL1- and NL2-mediated nuclear localization of the glucocorticoid receptor.

Molecular and cellular biology ·Vol. 19 ·No. 2 ·1999-02-00 ·Pages 1025-37

Savory JG, Hsu B, Laquian IR, Giffin W, Reich T, Haché RJ, Lefebvre YA

Abstract

Glucocorticoid receptor (GR) cycles between a free liganded form that is localized to the nucleus and a heat shock protein (hsp)-immunophilin-complexed, unliganded form that is usually localized to the cytoplasm but that can also be nuclear. In addition, rapid nucleocytoplasmic exchange or shuttling of the receptor underlies its localization. Nuclear import of liganded GR is mediated through a well-characterized sequence, NL1, adjacent to the receptor DNA binding domain and a second, uncharacterized motif, NL2, that overlaps with the ligand binding domain. In this study we report that rapid nuclear import (half-life [t1/2] of 4 to 6 min) of agonist- and antagonist-treated GR and the localization of unliganded, hsp-associated GRs to the nucleus in G0 are mediated through NL1 and correlate with the binding of GR to pendulin/importin alpha. By contrast, NL2-mediated nuclear transfer of GR occurred more slowly (t1/2 = 45 min to 1 h), was agonist specific, and appeared to be independent of binding to importin alpha. Together, these results suggest that NL2 mediates the nuclear import of GR through an alternative nuclear import pathway. Nuclear export of GR was inhibited by leptomycin B, suggesting that the transfer of GR to the cytoplasm is mediated through the CRM1-dependent pathway. Inhibition of GR nuclear export by leptomycin B enhanced the nuclear localization of both unliganded, wild-type GR and hormone-treated NL1(-) GR. These results highlight that the subcellular localization of both liganded and unliganded GRs is determined, at least in part, by a flexible equilibrium between the rates of nuclear import and export.

MeSH Terms
Amino Acid Sequence Amino Acid Substitution Animals Binding Sites/genetics Biological Transport, Active COS Cells Cell Nucleus Chloramphenicol O-Acetyltransferase/genetics Cytoplasm/metabolism Genes, Reporter Ligands Molecular Sequence Data Mutagenesis, Site-Directed Nuclear Proteins/metabolism Receptors, Glucocorticoid/chemistry,genetics,metabolism Recombinant Fusion Proteins/chemistry,genetics,metabolism Saccharomyces cerevisiae/genetics Transcriptional Activation Transfection
Chemicals
Ligands Nuclear Proteins Receptors, Glucocorticoid Recombinant Fusion Proteins pendulin Chloramphenicol O-Acetyltransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Savory J G
Department of Graduate Program in Biochemistry, The Loeb Health Research Institute at the Ottawa Hospital, University of Ottawa, Ottawa, Ontario, Canada.
Hsu B
Laquian I R
Giffin W
Reich T
Haché R J
Lefebvre Y A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.19.2.1025
Published
1999-02-00
Pages
1025-37
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC116033
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.19.2.1025 PMC Full Text View journal details
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