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α 1 ‐adrenergic activation of phospholipase D in CCL39 fibroblasts stimulates PLD1 via PKC and RhoA kinase
Author(s) -
Rastedt Danielle E,
Provost Joseph,
Wallert Mark A
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.a977
Subject(s) - phospholipase d , pld2 , rhoa , protein kinase c , phosphatidic acid , stimulation , chemistry , phosphorylation , agonist , junb , microbiology and biotechnology , receptor , signal transduction , endocrinology , biology , biochemistry , gene expression , phospholipid , membrane , gene
Phospholipase D (PLD) catalyzes the formation of the bioactive lipid, phosphatidic acid (PA). While the existence of two PLD isoforms has been identified, the role for each in cell signaling is much less well defined. Earlier studies in our laboratory have shown that PLD is upstream of ERK, NHE and MMP9 in α 1 ‐adrenergic receptor signaling regulating cell migration in CCL39 fibroblasts. In this study we show that cells treated with 50 μM phenylephrine (PE) a specific α 1 ‐ adrenergic receptor agonist, resulted in a 2.5 fold increase in phosphatidylbutanol (PtdBut) formation over unstimulated control cells. Expression of either PLD1 or PLD2 wild‐type both resulted in a minor increase in PLD activity, whereas expression of dominant negative PLD1 but not PLD2 abrogated PE mediated increases in the formation of PtdBut. Activation of PKC by the addition of 100 nm phorbal myristate ester resulted in a four‐fold increase in PLD activity indicating that PLD was stimulated by PKC. The PKC inhibitor Ro‐31‐8220 blocked PE stimulation of PLD. Further evidence that PLD1 isoform is activated by PE addition was shown when10 μM Y‐27632 blocked PtdBut formation after PE stimulation. Additional evidence indicating PLD1 is activated by the α 1 ‐ adrenergic receptor is shown by expression of dominant negative RhoA and Rock in PE stimulated cells. This study was supported with funds by NIH 1 R15 HL074924 ‐01A1