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J Biol Chem, Vol. 274, Issue 6, 3414-3421, February 5, 1999
From the A central function of cystic fibrosis
transmembrane conductance regulator (CFTR)-expressing tissues is the
secretion of fluid containing 100-140 mM
HCO3
Regulation of
Cl
/ HCO3
Exchange by Cystic Fibrosis Transmembrane Conductance Regulator
Expressed in NIH 3T3 and HEK 293 Cells
,,,,, and
Department of Physiology, University of
Texas Southwestern Medical Center, Dallas, Texas 75235 and the
¶ Department of Medicine and Physiology, University of Tennessee,
Memphis, Tennessee 38163
. High levels of HCO3 maintain
secreted proteins such as mucins (all tissues) and digestive enzymes
(pancreas) in a soluble and/or inactive state. HCO3
secretion is impaired in CF in all CFTR-expressing,
HCO3-secreting tissues examined. The mechanism
responsible for this critical problem in CF is unknown. Since a major
component of HCO3 secretion in CFTR-expressing cells
is mediated by the action of a Cl/HCO3
exchanger (AE), in the present work we examined the regulation of AE
activity by CFTR. In NIH 3T3 cells stably transfected with wild type
CFTR and in HEK 293 cells expressing WT and several mutant CFTR,
activation of CFTR by cAMP stimulated AE activity. Pharmacological and
mutagenesis studies indicated that expression of CFTR in the plasma
membrane, but not the Cl conductive function of CFTR was
required for activation of AE. Furthermore, mutations in NBD2 altered
regulation of AE activity by CFTR independent of their effect on
Cl channel activity. At very high expression levels CFTR
modified the sensitivity of AE to
4,4'-diisothiocyanatostilbene-2,2'-disulfonate. The novel finding of
regulation of Cl/HCO3 exchange by CFTR
reported here may have important physiological implications and
explain, at least in part, the impaired HCO3
secretion in CF.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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