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J Biol Chem, Vol. 273, Issue 48, 32304-32311, November 27, 1998
From the Departments of Shear stress, the dragging force generated
by fluid flow, differentially activates extracellular signal-regulated
kinase (ERK) and c-Jun NH2-terminal kinase (JNK) in
bovine aortic endothelial cells (BAEC) (Jo, H., Sipos, K., Go, Y. M., Law, R., Rong, J., and McDonald, J. M. (1997) J. Biol. Chem. 272, 1395-1401). Here, we examine whether
cholesterol-enriched compartments in the plasma membrane are
responsible for such differential regulation. Pretreatment of BAEC with
a cholesterol-binding antibiotic, filipin, did not inhibit
shear-dependent activation of JNK. In contrast, filipin and
other membrane-permeable cholesterol-binding agents (digitonin and
nystatin), but not the lipid-binding agent xylazine, inhibited shear-dependent activation of ERK. The effect of
cholesterol-binding drugs did not appear to be due to membrane
permeabilization, since treatment of BAEC with a detergent, Triton
X-100 which also permeabilizes membranes, did not inhibit
shear-dependent activation of ERK. Furthermore,
shear-dependent activation of ERK, but not JNK, was inhibited by cyclodextrin, a membrane-impermeable cholesterol-binding agent, which removes cell-surface cholesterol. Moreover, the effects of
cyclodextrin were prevented by adding cholesterol during the incubation. These results indicate that cholesterol or
cholesterol-sensitive compartments in the plasma membrane play a
selective and essential role in activation of ERK, but not JNK, by
shear stress. Although exposure to shear stress (1 h) increased the
number of caveolae by 3-fold, treatment with filipin had no effect in
either control or shear-exposed cells suggesting that caveolae density
per se is not a crucial determinant in
shear-dependent ERK activation. In summary, the current
study suggests that cholesterol-sensitive microdomains in the plasma
membrane, such as caveolae-like domains, play a critical role in
differential activation of ERK and JNK by shear stress.
Plasma Membrane Cholesterol Is a Key Molecule in Shear
Stress-dependent Activation of Extracellular
Signal-regulated Kinase
,,,
Pathology and
§ Cell Biology, University of Alabama at Birmingham,
Birmingham, Alabama 35294 and the ¶ Department of Molecular
Pharmacology, Albert Einstein College of Medicine, Bronx, New
York 10461
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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