Participation of phospholipase D and α/β-protein kinase C in growth factor-induced signalling in C3H10T1/2 fibroblasts
| Autor: | M. Vorland, Ove Bruland, Holm Holmsen, Johan R. Lillehaug, Bodil Bjørndal, Vidar A.T. Thorsen |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
MAPK/ERK pathway
Platelet-derived growth factor Swine Phosphatidic Acids Glycerophospholipids Biology Cell Line Mice chemistry.chemical_compound 1-Butanol Epidermal growth factor Phospholipase D Animals RNA Messenger Enzyme Inhibitors Growth Substances Molecular Biology Protein Kinase C Protein kinase C Platelet-Derived Growth Factor Mice Inbred C3H Mitogen-Activated Protein Kinase 3 Epidermal Growth Factor Kinase PLD2 Receptor for activated C kinase 1 Cell Biology Fibroblasts Molecular biology Cell biology Enzyme Activation chemistry Tetradecanoylphorbol Acetate lipids (amino acids peptides and proteins) Mitogen-Activated Protein Kinases Peptides Proto-Oncogene Proteins c-fos Signal Transduction |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids. 1632:62-71 |
| ISSN: | 1388-1981 |
| DOI: | 10.1016/s1388-1981(03)00063-5 |
| Popis: | We have studied phospholipase D (PLD) activation in relation to protein kinase C (PKC) and the involvement of PLD in extracellularly regulated kinase 1 (MAPK) (ERK1) activation and c-fos mRNA expression in C3H/10T1/2 (Cl8) fibroblasts. In these cells, the PLD activity was significantly increased by porcine platelet-derived growth factor (PDGF-BB), phorbol 12-myristate 13-acetate (PMA), and epidermal growth factor (EGF). PLD activation by PDGF-BB and PMA, but not EGF, was inhibited in Cl8 cells expressing the HAbetaC2-1 peptide (Cl8 HAbetaC2-1 cells), with a sequence (betaC2-1) shown to bind receptor for activated C kinase 1 (RACK1) and inhibit c-PKC-mediated cell functions [Science 268 (1995) 247]. A role of alpha-PKC in PLD activation is further underscored by co-immunoprecipitation of alpha-PKC with PLD1 and PLD2 in non-stimulated as well as PMA- and PDGF-BB-stimulated Cl8 cells. However, only PKC in PLD1 precipitates was activated by these agonists, while the PKC in the PLD2 precipitates was constitutively activated. The c-fos mRNA levels in Cl8 cells increased more than 30-fold in response to either PDGF-BB, EGF, or PMA. Approximately 60% inhibition of this increase in c-fos mRNA levels was observed in Cl8 HAbetaC2-1 cells. Formation of phosphatidylbutanol (PtdBut) at the expense of phosphatidic acid (PtdH) in the presence of n-butanol inhibited ERK1 activation and c-fos mRNA expression in PDGF-BB-treated Cl8 cells. ERK activation by PMA was unaffected by n-butanol in Cl8 cells but almost abolished by n-butanol in Cl8 HAbetaC2-1 cells, showing that ERK activation by PMA is heavily dependent on PKC and PLD1. In contrast, ERK activation by EGF in both cell types was not sensitive to n-butanol. These results indicate (1) a role of a functional interaction between the RACK1 scaffolding protein and a alphaPKC-PLD complex for achieving full PLD activity in PDGF-BB- and PMA-stimulated Cl8 cells; (2) PLD-mediated PtdH formation is needed for optimal ERK1 activation by PDGF-BB and maximal increase in c-fos mRNA expression. These findings place PLD as an important component in PDGF-BB- and PMA-stimulated intracellular signalling leading to gene activation in Cl8 cells, while EGF does not require PLD. |
| Databáze: | OpenAIRE |
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