scholarly journals Stimulation of the hydrolysis of phosphatidylinositol 4,5-bisphosphate and phosphatidylcholine by endothelin, a complete mitogen for Rat-1 fibroblasts

1990 ◽  
Vol 272 (3) ◽  
pp. 761-766 ◽  
Author(s):  
E E MacNulty ◽  
R Plevin ◽  
M J O Wakelam
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Endothelin 1 ◽  
Kinase C ◽  
Choline Phosphate ◽  
Rapid Generation ◽  
Endothelin 3 ◽  
Hydrolysis Of ◽  
Phosphatidylcholine Synthesis ◽  
Stimulation Of

The mitogenic activity of endothelin and its ability to stimulate PtdIns(4,5)P2 and phosphatidylcholine turnover in Rat-1 fibroblasts was studied. Stimulated incorporation of [3H]thymidine occurred in the absence of any other added growth factors. The endothelins stimulated rapid generation of both Ins(1,4,5)P3 and choline. Endothelin-1 and endothelin-2 were equipotent in stimulating both responses, but endothelin-3 was less potent. Endothelin-1-stimulated Ins(1,4,5)P3 generation reached a maximum at 5 s and then declined; however, the response was long-lived, with a 4.5-fold elevation over basal still observed after 15 min. Endothelin-stimulated choline generation was observed with no increase in choline phosphate; indeed, the apparent level of this metabolite fell after 30 min of stimulation, presumably due to the observed stimulation of phosphatidylcholine synthesis. The endothelin-stimulated increase in choline generation was abolished in cells where protein kinase C was down-regulated. However, endothelin-stimulated choline generation was greater than that observed in response to a protein kinase C-activating phorbol ester, raising the possibility that the peptide activates phospholipase D by both protein kinase C-dependent and -independent mechanisms.

scholarly journals Analysis of the water-soluble products of phosphatidylcholine breakdown by ion-exchange chromatography. Bombesin and TPA (12-O-tetradecanoylphorbol 13-acetate) stimulate choline generation in Swiss 3T3 cells by a common mechanism

1989 ◽  
Vol 263 (2) ◽  
pp. 581-587 ◽  
Author(s):  
S J Cook ◽  
M J O Wakelam
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Ion Exchange ◽  
Water Soluble ◽  
Common Mechanism ◽  
3T3 Cells ◽  
Kinase C ◽  
Choline Phosphate ◽  
Swiss 3T3 ◽  
Stimulation Of

A method for the rapid and quantitative separation of glycerophosphocholine, choline phosphate and choline upon ion-exchange columns is described. The method has been utilized to examine the stimulation of phosphatidylcholine breakdown in quiescent Swiss 3T3 cells in response to bombesin and 12-O-tetradecanoylphorbol 13-acetate (TPA). The stimulated generation of choline is shown to precede that of choline phosphate, with no effect upon glycerophosphocholine levels; but was attenuated in cells in which protein kinase C activity was down-regulated. The results thus suggest that stimulation of the cells with either bombesin or TPA activates phospholipase D-catalysed phosphatidylcholine breakdown by a common mechanism involving the activation of protein kinase C.

Stimulation of catecholamine biosynthesis via the protein kinase C pathway by endothelin-1 in PC12 rat pheochromocytoma cells

2002 ◽  
Vol 63 (5) ◽  
pp. 977-984 ◽  
Author(s):  
Kazuhiro Takekoshi ◽  
Kiyoaki Ishii ◽  
Shunsuke Shibuya ◽  
Yasushi Kawakami ◽  
Kazumasa Isobe ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Catecholamine Biosynthesis ◽  
Pheochromocytoma Cells ◽  
Endothelin 1 ◽  
Kinase C ◽  
Stimulation Of

scholarly journals Endothelin-1 stimulates DNA synthesis and anchorage-independent growth of Rat-1 fibroblasts through a protein kinase C-dependent mechanism.

1990 ◽  
Vol 1 (4) ◽  
pp. 379-390 ◽  
Author(s):  
L L Muldoon ◽  
D Pribnow ◽  
K D Rodland ◽  
B E Magun
Keyword(s):  
Cell Proliferation ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Dna Synthesis ◽  
Second Messenger ◽  
Anchorage Independent Growth ◽  
Endothelin 1 ◽  
Essential Component ◽  
Kinase C ◽  
Stimulation Of

Stimulation of quiescent cultured fibroblasts with a variety of growth-promoting factors induces release of diacylglycerol (DG) and subsequent activation of protein kinase C (pkC), but the role of pkC in the induction of DNA synthesis and cell proliferation remains unclear. We have investigated the involvement of pkC in the response of Rat-1 fibroblasts to the newly described peptide endothelin-1 (Et-1), an agonist that is secreted by the vascular endothelium and that may play a role in the proliferative response of cells in the vessel wall. Addition of Et-1 to serum-deprived Rat-1 cells promoted DNA synthesis in the absence of additional factors and stimulated anchorage-independent growth in the presence of epidermal growth factor (EGF), indicating that Et-1 has many of the characteristics of a mitogen. The ability of Et-1 to stimulate both DNA synthesis and anchorage-independent growth was markedly reduced by the depletion of cellular pkC activity induced by prolonged exposure to 12-O-tetradecanoylphorbol-13-acetate (TPA). In contrast, the ability of Et-1 to induce both second messenger production and transcription of c-fos and c-jun was largely independent of cellular pkC activity. Production of DG in response to Et-1 persisted for greater than 12 h and may account for the ability of Et-1 to augment the G1-S phase transition. Although these observations indicate that functional pkC is not an essential component of the proximal pathway leading to rapid changes in gene transcription and second messenger production in response to Et-1 treatment, the data suggest that activation of pkC is an essential component of the downstream events responsible for the stimulation of cell proliferation and anchorage-independent growth in Rat-1 cells exposed to Et-1.

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