Differential Phospholipase D Activation by Bradykinin and Sphingosine 1-Phosphate in NIH 3T3 Fibroblasts Overexpressing Gelsolin

• Published in: The Journal of biological chemistry Vol. 274; no. 39; pp. 27385 - 27391
• Main Authors: Banno, Yoshiko, Fujita, Hisakazu, Ono, Yoshitaka, Nakashima, Shigeru, Ito, Yuzuru, Kuzumaki, Noboru, Nozawa, Yoshinori
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 24.09.1999; American Society for Biochemistry and Molecular Biology
• Subjects: 3T3 Cells; Animals; Bordetella pertussis; Bradykinin - pharmacology; Calcimycin - pharmacology; Cell Membrane - enzymology; DNA, Complementary; Egtazic Acid - pharmacology; Enzyme Activation; gelsolin; Gelsolin - genetics; Gelsolin - metabolism; Humans; Isoenzymes - metabolism; Kinetics; Lysophospholipids; MAP kinase; Mice; Mitogen-Activated Protein Kinases - metabolism; Pertussis Toxin; Phosphatidylinositol 4,5-Diphosphate - pharmacology; phospholipase D; Phospholipase D - metabolism; Protein Kinase C - metabolism; Protein Kinase C beta; Protein Kinase C-alpha; Protein Kinase C-epsilon; Recombinant Proteins - metabolism; Sphingosine - analogs & derivatives; Sphingosine - pharmacology; sphingosine 1-phosphate; Tetradecanoylphorbol Acetate - pharmacology; Transfection; Type C Phospholipases - metabolism; Virulence Factors, Bordetella - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.39.27385

Gelsolin, an actin-binding protein, shows a strong ability to bind to phosphatidylinositol 4,5-bisphosphate (PIP2). Here we showed in in vitroexperiments that gelsolin inhibited recombinant phospholipase D1 (PLD1) and PLD2 activities but not the oleate-dependent PLD and that this inhibition was not reversed by increasing PIP2concentration. To investigate the role of gelsolin in agonist-mediated PLD activation, we used NIH 3T3 fibroblasts stably transfected with the cDNA for human cytosolic gelsolin. Gelsolin overexpression suppressed bradykinin-induced activation of phospholipase C (PLC) and PLD. On the other hand, sphingosine 1-phosphate (S1P)-induced PLD activation could not be modified by gelsolin overexpression, whereas PLC activation was suppressed. PLD activation by phorbol myristate acetate or Ca2+ ionophore A23187 was not affected by gelsolin overexpression. Stimulation of control cells with either bradykinin or S1P caused translocation of protein kinase C (PKC) to the membranes. Translocation of PKC-α and PKC-β1 but not PKC-ε was reduced in gelsolin-overexpressed cells, whereas phosphorylation of mitogen-activated protein kinase was not changed. S1P-induced PLC activation and mitogen-activated protein kinase phosphorylation were sensitive to pertussis toxin, but PLD response was insensitive to such treatment, suggesting that S1P induced PLD activation via certain G protein distinct from Gi for PLC and mitogen-activated protein kinase pathway. Our results suggest that gelsolin modulates bradykinin-mediated PLD activation via suppression of PLC and PKC activities but did not affect S1P-mediated PLD activation.