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Role of phospholipase D in α 1 ‐adrenergic receptor signaling in CCL39 fibroblasts
Author(s) -
Nguyen Callie,
Taves Jennifer,
Mork David,
Wallert Mark A,
Provost Joseph J
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a976-d
Subject(s) - phospholipase d , phosphatidic acid , mapk/erk pathway , rhoa , phosphorylation , stress fiber , microbiology and biotechnology , motility , chemistry , stimulation , signal transduction , cell growth , focal adhesion , biology , biochemistry , endocrinology , phospholipid , membrane
Phospholipase D (PLD) has been implicated in the progression of tumorgenesis and cell migration. We have found that stimulation of the α 1 ‐adrenergic receptor ( AR) in CCL39 fibroblasts leads to both cell migration and PLD activity. Addition of 50 μM PE resulted to a two fold increase in ERK phosphorylation, a dramatic increase in Ras activation and a three fold increase in the number of cells displaying strong stress fiber formation. Each event is critical for cell motility in fibroblasts. To investigate the role of PLD, we used alcohol to inhibit the ability of PLD to form its product, PA. Addition of 1‐butanol inhibited ERK phosphorylation and stress fiber formation following stimulation by PE. Lower concentrations (<0.4%v/v) of 2‐butanol did not significantly inhibit PE mediated ERK and stress fiber activation. Chemical inhibition of PKC or RhoA Kinase both, reduced the amount of ERK phosphorylation by PE. Finally to show the role of PA in mediating this response, we use short chain PA to act as a cell permeable second messenger of PLD. 40μM short chain phosphatidic acid (scPA) dramatically increased the amounts of activated Ras near the levels of PE alone and 80μM scPA increased the amounts of activated Ras twice that of the 40μM scPA. These results indicate that the AR acts through PLD to activate the ERK growth factor pathway and points to a new role for PLD in mediating cell motility events. NIH 5‐R01‐HL07924 & NIH URS