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J. Biol. Chem., Vol. 277, Issue 21, 18598-18604, May 24, 2002
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From the Laboratoire de Biologie Intégrée des Cellules
Rénales, Service de Biologie Cellulaire, Commissariat á
l'Energie Atomique, Saclay, Unité de Recherche Associée
1859, CNRS, 91191 Gif-sur-Yvette Cedex, France
This study aimed at determining the signaling
pathways underlying calcitonin- and isoproterenol-induced stimulation
of H,K-ATPase in rat renal collecting duct. H,K-ATPase activity was
determined in microdissected collecting ducts preincubated with or
without either specific inhibitors or antibodies directed against
intracellular signaling proteins. Transient cell membrane
permeabilization with streptolysin-O allowed intracellular access of
antibodies. The stimulation of H,K-ATPase by calcitonin and
isoproterenol was mimicked by cAMP analogues and was abolished by
adenylyl cyclase inhibition. Protein kinase A inhibition abolished
isoproterenol but not calcitonin effect on H,K-ATPase. Calcitonin
increased the phosphorylation of extracellular signal-regulated kinase
(ERK) in a protein kinase A-independent manner, and the inhibition of the ERK phosphorylation prevented the stimulation of H,K-ATPase by
calcitonin. Antibodies directed against either the cAMP-activated guanine-nucleotide exchange factor Epac I, the monomeric G protein Rap-1 or the kinase Raf-B, curtailed the stimulation of H,K-ATPase by
calcitonin, whereas antibodies against the related monomeric G protein
Ras or kinase Raf-1 had no effect. In conclusion, calcitonin stimulates
H,K-ATPase through a cAMP/Epac I/Rap-1/Raf-B/ERK cascade.
Protein Kinase A-independent Activation of ERK and H,K-ATPase by
cAMP in Native Kidney Cells
ROLE OF Epac I*
*
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: URA 1859, Bâtiment 520, Centre d'Etudes de Saclay, 91191 Gif-sur-Yvette
cedex, France. Tel.: 33-169089761; Fax: 33-169083570; E-mail:
doucet@dsvidf.cea.fr.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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