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J Biol Chem, Vol. 275, Issue 11, 8133-8142, March 17, 2000
From the Na+/H+ exchanger
NHE3 is a plasma membrane (PM) protein, which contributes to
Na+ absorption in the intestine. Growth factors stimulate
NHE3 via phosphatidylinositol 3-kinase (PI3-K), but mechanism of this
process is not clear. To examine the hypothesis that growth factors
stimulate NHE3 by modulating NHE3 recycling, and that PI3-K
participates in this mechanism, we used PS120 fibroblasts expressing a
fusion protein of NHE3 and green fluorescent protein. At steady state, ~25% of cellular NHE3 content was expressed at PM. Inhibition of
PI3-K decreased PM expression of NHE3, which correlated with retention
of the exchanger in recycling endosomal compartment. In contrast, basic
fibroblast growth factor (bFGF) increased PM expression of NHE3, which
was associated with a 2-fold increase in rate constant for exit of the
exchanger from the recycling compartment. Qualitatively similar effects
of bFGF were observed in cells pretreated with PI3-K inhibitors, but
their magnitude was only ~50% of that in intact cells. These data
suggest that: (i) bFGF stimulates NHE3 by increasing PM expression of
the exchanger; (ii) PI3-K mediates PM expression of NHE3 in both basal
and bFGF-stimulated conditions, and (iii) not all of the effects of
bFGF on NHE3 expression are mediated by PI3-K, suggesting additional
regulatory mechanisms.
Basic Fibroblast Growth Factor Stimulates Surface Expression and
Activity of Na+/H+ Exchanger NHE3 via Mechanism
Involving Phosphatidylinositol 3-Kinase*
§¶, Department of Medicine, Division of
Gastroenterology, Hepatology, and Nutrition, University of Texas
Medical School, Houston, Texas 77030 and the § Departments
of Medicine and Physiology, Division of Gastroenterology, The Johns
Hopkins University School of Medicine, Baltimore, Maryland 21205
*
This work was supported by National Institutes of Health
NIDDK Grants K08DK02557, RO1DK26523, PO1DK44484, R29DK43778, and T32DK0763205, and by the Meyerhoff Digestive Diseases Center for Epithelial Disorders. Part of this work was presented at the 100th Annual Meeting of the American Gastroenterological Association, Orlando, FL, May 16-19, 1999 (Abstract 3877), and was published in
abstract form (Janecki, A. J., Janecki, M., Akhter, S., and Donowitz,
M. (1999) Gastroenterology 116, G3877).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.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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