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J Biol Chem, Vol. 273, Issue 34, 21542-21553, August 21, 1998
From the a Molecular Medicine and Renal Units, Beth Israel
Deaconess Medical Center Boston, Massachusetts 02215, the
e Division of Hematology-Oncology, Beth Israel Deaconess Medical
Center and Harvard Institutes of Medicine, Boston, Massachusetts 02215, the g Department of Obstetrics and Gynecology and
h Vollum Institute, Oregon Health Sciences University, Portland,
Oregon 97201, the i Departments of Internal Medicine and General
Pathology, University of Verona, 37100 Verona, Italy, the
j Department of Internal Medicine, Maggiore Hospital IRCCS,
University of Milan, 20122 Milan, Italy, the k Department of
Laboratory Medicine, Children's Hospital, Boston, Massachusetts 02115, and the Departments of b Medicine, l Pathology, and
m Cell Biology, Harvard Medical School,
Boston, Massachusetts 02115
We have cloned from murine
erythroleukemia (MEL) cells, thymus, and stomach the cDNA encoding
the Ca2+-gated K+ (KCa)
channel, mIK1, the mouse homolog of hIK1 (Ishii, T. M., Silvia,
C., Hirschberg, B., Bond, C. T., Adelman, J. P., and Maylie, J. (1997) Proc. Natl. Acad. Sci.(U. S. A. 94, 11651-11656). mIK1 mRNA was detected at varied levels in many
tissue types. mIK1 KCa channel activity expressed in
Xenopus oocytes closely resembled the Kca of
red cells (Gardos channel) and MEL cells in its single channel
conductance, lack of voltage-sensitivity of activation, inward
rectification, and Ca2+ concentration dependence. mIK1 also
resembled the erythroid channel in its pharmacological properties,
mediating whole cell and unitary currents sensitive to low
nM concentrations of both clotrimazole (CLT) and its
des-imidazolyl metabolite, 2-chlorophenyl-bisphenyl-methanol, and to
low nM concentrations of iodocharybdotoxin. Whereas control oocytes subjected to hypotonic swelling remained swollen, mIK1 expression conferred on oocytes a novel,
Ca2+-dependent, CLT-sensitive regulatory volume
decrease response. Hypotonic swelling of voltage-clamped
mIK1-expressing oocytes increased outward currents that were
Ca2+-dependent, CLT-sensitive, and reversed
near the K+ equilibrium potential. mIK1 mRNA levels in
ES cells increased steadily during erythroid differentiation in
culture, in contrast to other KCa mRNAs examined. Low
nanomolar concentrations of CLT inhibited proliferation and erythroid
differentiation of peripheral blood stem cells in liquid culture.
cDNA Cloning and Functional Characterization of the Mouse
Ca2+-gated K+ Channel, mIK1
ROLES IN REGULATORY VOLUME DECREASE AND ERYTHROID
DIFFERENTIATION
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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