Role of Ras-dependent ERK activation in phorbol ester-induced endothelial cell barrier dysfunction

Alexander D. Verin, Feng Liu, Natalia Bogatcheva, Talaibek Borbiev, Marc B. Hershenson, Peiyi Wang, Joe GN Garcia

Research output: Contribution to journalArticle

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Abstract

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.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume279
Issue number2 23-2
StatePublished - 2000
Externally publishedYes

Fingerprint

Phorbol Esters
Acetates
Endothelial Cells
Protein Kinase C
Mitogen-Activated Protein Kinase Kinases
Farnesyltranstransferase
MAP Kinase Kinase Kinases
Antisense Oligonucleotides
Extracellular Signal-Regulated MAP Kinases
Electric Impedance
phorbol-12-myristate
Phosphotransferases

Keywords

  • Cytoskeleton
  • Extracellular signal-regulated kinase
  • Mitogen-activated protein kinases

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology
  • Physiology (medical)

Cite this

Role of Ras-dependent ERK activation in phorbol ester-induced endothelial cell barrier dysfunction. / Verin, Alexander D.; Liu, Feng; Bogatcheva, Natalia; Borbiev, Talaibek; Hershenson, Marc B.; Wang, Peiyi; Garcia, Joe GN.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 279, No. 2 23-2, 2000.

Research output: Contribution to journalArticle

Verin, Alexander D. ; Liu, Feng ; Bogatcheva, Natalia ; Borbiev, Talaibek ; Hershenson, Marc B. ; Wang, Peiyi ; Garcia, Joe GN. / Role of Ras-dependent ERK activation in phorbol ester-induced endothelial cell barrier dysfunction. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2000 ; Vol. 279, No. 2 23-2.
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