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J. Biol. Chem., Vol. 278, Issue 19, 17320-17327, May 9, 2003
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From the PSD-95/Dlg-A/ZO-1 (PDZ) domains play an
essential role in determining cell polarity. The
Na+/H+ exchanger regulatory factor
(NHERF), also known as EBP50, contains two PDZ domains that mediate the
assembly of transmembrane and cytosolic proteins into functional signal
transduction complexes. Moreover, it has been shown that cystic
fibrosis transmembrane conductance regulator (CFTR) and
Stimulation of
2-Adrenergic Receptor Increases
Cystic Fibrosis Transmembrane Conductance Regulator Expression
in Human Airway Epithelial Cells through a cAMP/Protein Kinase
A-independent Pathway*
,,,
INSERM 514, IFR 53, Centre
Hospitalier Universitaire Maison Blanche, Reims, 51092 Cedex, France,
§ Hopital Tenon, 4 rue de la Chine, 75020 Paris, France, and
¶ Hopital Jean Bernard, BP 577, 86021 Poitiers, France
2-adrenergic receptor (2AR) bind equally
well to the PDZ1 domain of EBP50. We hypothesized that
2AR activation may regulate CFTR protein expression. To verify this, we evaluated the effects of a pharmacologically relevant concentration of salmeterol (2.10
7 M), a long
acting 2AR agonist, on CFTR expression in primary human
airway epithelial cells (HAEC). 2AR stimulation induced a time-dependent increase in apical CFTR protein
expression, with a maximal response reached after treatment for 24 h. This effect was post-transcriptional, dependent upon the
2AR agonist binding to 2AR and
independent of the known 2AR agonist-mediated cAMP/PKA pathway. We demonstrated by immunohistochemistry that CFTR,
2AR, and EBP50 localize to the apical membrane of HAEC.
Analyses of anti-EBP50 protein immunoprecipitate showed that salmeterol
induced an increase in the amount of CFTR that binds to EBP50. These
data suggest that 2AR activation regulates the
association of CFTR with EBP50 in polarized HAEC.
*
This work was supported by INSERM, GlaxoSmithKline,
and by the Association Vaincre la Mucoviscidose.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: INSERM
514, Centre Hospitalier Universitaire Maison Blanche 45, rue Cognacq
Jay, 51092 Reims Cedex, France. Tel.: 33-3-26-78-77-70; Fax:
33-3-26-06-58-01; E-mail: epuche@worldnet.fr.
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