Activation of phospholipase D in CHO cells transfected with the human epidermal growth factor (EGF) receptor: differential effects of protein kinase C activation and EGF.
Multiple intracellular signal transduction pathways, including phospholipases A2 and D, can be activated by epidermal growth factor (EGF) in both a protein kinase C (PKC)-dependent and -independent manner. We investigated the activation of phospholipase D (PLD) by a PKC activator, phorbol myristate acetate (PMA) and by EGF in CHO cells transfected with the full-length EGF receptor. In cells labelled with arachidonic acid or linoleic acid, PMA activated a PLD, determined by formation of the transphosphatidylation product phosphatidylethanol in the presence of ethanol. A basal PLD activity was seen in linoleic acid-labelled cells but not in cells labelled with arachidonic acid. This basal activity was augmented by the protein phosphotyrosine phosphatase inhibitor vanadate and reduced by tyrosine kinase inhibition and was contributed to by PKC, as activity could not be elicited following prolonged exposure to phorbol ester, known to down-regulate some PKC isoforms. By contrast, EGF failed to stimulate formation of phosphatidylethanol in cells labelled with either fatty acid species. It is proposed that in the basal condition PKC-dependent PLD activation and protein tyrosine kinase phosphorylation are linked (possibly by a phospholipase C (PLC)-mediated formation of diacylglycerol); EGF which activated a phospholipase A2 (PLA2) but which failed to elicit PLC activation in these cells is without further effect on PLD.
Dunlop
M
M
University of Melbourne Department of Medicine, Royal Melbourne Hospital, Parkville, Victoria, Australia.
Clark
S
S
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000378:metabolism*; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D010739:Phospholipase D / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D010743:Phospholipids; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D011493:Protein Kinase C; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate