TY - JOUR
T1 - Role of Ras-dependent ERK activation in phorbol ester-induced endothelial cell barrier dysfunction
AU - Verin, Alexander Dmitriyevich
AU - Liu, Feng
AU - Bogatcheva, Natalia
AU - Borbiev, Talaibek
AU - Hershenson, Marc B.
AU - Wang, Peiyi
AU - Garcia, Joe G.N.
PY - 2000
Y1 - 2000
N2 - The treatment of endothelial cell monolayers with phorbol 12-myristate 13-acetate (PMA), a direct protein kinase C (PKC) activator, leads to disruption of endothelial cell monolayer integrity and intercellular gap formation. Selective inhibition of PKC (with bisindolylmaleimide) and extracellular signal-regulated kinases (ERKs; with PD-98059, olomoucine, or ERK antisense oligonucleotides) significantly attenuated PMA-induced reductions in transmonolayer electrical resistance consistent with PKC- and ERK-mediated endothelial cell barrier regulation. An inhibitor of the dual-specificity ERK kinase (MEK), PD-98059, completely abolished PMA-induced ERK activation. PMA also produced significant time-dependent increases in the activity of Raf-1, a Ser/Thr kinase known to activate MEK (~6-fold increase over basal level). Similarly, PMA increased the activity of Ras, which binds and activates Raf-1 (~80% increase over basal level). The Ras inhibitor farnesyltransferase inhibitor III (100 μM for 3 h) completely abolished PMA-induced Raf-1 activation. Taken together, these data suggest that the sequential activation of Ras, Raf-1, and MEK are involved in PKC-dependent endothelial cell barrier regulation.
AB - The treatment of endothelial cell monolayers with phorbol 12-myristate 13-acetate (PMA), a direct protein kinase C (PKC) activator, leads to disruption of endothelial cell monolayer integrity and intercellular gap formation. Selective inhibition of PKC (with bisindolylmaleimide) and extracellular signal-regulated kinases (ERKs; with PD-98059, olomoucine, or ERK antisense oligonucleotides) significantly attenuated PMA-induced reductions in transmonolayer electrical resistance consistent with PKC- and ERK-mediated endothelial cell barrier regulation. An inhibitor of the dual-specificity ERK kinase (MEK), PD-98059, completely abolished PMA-induced ERK activation. PMA also produced significant time-dependent increases in the activity of Raf-1, a Ser/Thr kinase known to activate MEK (~6-fold increase over basal level). Similarly, PMA increased the activity of Ras, which binds and activates Raf-1 (~80% increase over basal level). The Ras inhibitor farnesyltransferase inhibitor III (100 μM for 3 h) completely abolished PMA-induced Raf-1 activation. Taken together, these data suggest that the sequential activation of Ras, Raf-1, and MEK are involved in PKC-dependent endothelial cell barrier regulation.
KW - Cytoskeleton
KW - Extracellular signal-regulated kinase
KW - Mitogen-activated protein kinases
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U2 - 10.1152/ajplung.2000.279.2.l360
DO - 10.1152/ajplung.2000.279.2.l360
M3 - Article
C2 - 10926560
AN - SCOPUS:0033848682
SN - 1040-0605
VL - 279
SP - L360-L370
JO - American Journal of Physiology - Lung Cellular and Molecular Physiology
JF - American Journal of Physiology - Lung Cellular and Molecular Physiology
IS - 2 23-2
ER -