A role of p44/42 mitogen-activated protein kinases in formyl-peptide receptor-mediated phospholipase D activity and oxidant production.
Phosphatidylcholine-specific phospholipase D (PLD) is a major cellular source of phosphatidic acid and choline, which regulate various physiopathological processes. PLD activation mediated by chemoattractants involves protein phosphorylation. This study provides pharmacological and biochemical evidence of a major role of p44/42 MAP kinases (ERK1/2) in PLD activation induced by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP). ERK1/2 inhibition by the MEK1/2 antagonist U0126 in neutrophilic HL-60 cells or HEK 293T cells stably expressing fMLP receptors abolished fMLP-mediated PLD activity. Conversely, a constitutively activated MEK1 mutant expressed in HEK 293T cells potentiated fMLP-induced PLD activity. Expression of inactive PLD mutants showed that PLD2, but not PLD1, contributed to fMLP-mediated PLD activity. PLD2 co-immunoprecipitated with ERK1/2 and became phosphorylated on MAP kinase consensus sites in fMLP-stimulated cells. In cell-free systems, ERK2 gave rise to strong ATP-dependent PLD activity and directly phosphorylated PLD2 that generated two phosphopeptides only after tryptic digestion. Finally, pharmacological inhibition of ERK activation and the inhibition of PLD expression by antisense oligonucleotides in HL-60 cells suggest that the ERK/PLD2 pathway contributes to fMLP-mediated oxidant production. In conclusion, the fMLP-mediated PLD activity is regulated by ERK1/2, involving a predominant contribution of PLD2. The ERK/PLD2 coupling may provide potential pharmacological targets to control PLD-associated cellular dysfunctions.
Paruch
Sylvain
S
Département de Biologie Cellulaire de l'Institut Cochin, The Institut National de la Recherche Médicale (INSERM U567), The Centre National de la Recherche Scientifique (UMR 8104), and the Université René Descartes, Paris France.
El-Benna
Jamel
J
Djerdjouri
Bahia
B
Marullo
Stéfano
S
Périanin
Axel
A
D002070:Butadienes; D002460:Cell Line; D002633:Chemotaxis; D006801:Humans; D007527:Isoenzymes; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009154:Mutation; D009570:Nitriles; D016877:Oxidants / Q000096:biosynthesis*; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D044042:Receptors, Formyl Peptide / Q000378:metabolism*; D016897:Respiratory Burst
D002070:Butadienes; D007527:Isoenzymes; D009570:Nitriles; D016877:Oxidants; D044042:Receptors, Formyl Peptide; C113580:U 0126; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D010739:Phospholipase D