title: Activation of membrane-bound phospholipase D by protein kinase C in HL60 cells: synergistic action of a small GTP-binding protein RhoA.
abstract: Regulation of phospholipase D (PLD) activation by protein kinase C (PKC) was studied in membranes isolated from human promyelocytic leukemia HL60 cells. The activation of membrane-bound PLD by PKC partially purified from rat brain was most effectively induced with phorbol 12-myristate 13-acetate (PMA) and Ca2+ (1 microM) which caused translocation of PKC to membranes. Ro31-8425, a potent inhibitor of PKC, suppressed the catalytic activity of PKC in a concentration-dependent manner, with complete inhibition at 5 microM. However, the PKC-mediated PLD activation was not affected by Ro31-8425. It was thus suggested that membrane-bound PLD of HL60 cells was activated by PKC translocation but probably via a phosphorylation-independent mechanism. Furthermore, addition by guanosine 5'-3-O-(thio)trisphosphate (GTP gamma S) potentiated the PKC-mediated PLD activation and this potentiating effect was abolished by Rho GTPase dissociation inhibitor (RhoGDI). The suppressed PLD activation by RhoGDI was completely restored by addition of recombinant RhoA. These results indicate that the PKC-mediated PLD activation can be synergistically potentiated by RhoA in HL60 membranes.
authors.affiliation: Department of Biochemistry, Gifu University School of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ohguchi
authors.forename: Y
authors.initials: Y
authors.lastname: Banno
authors.forename: S
authors.initials: S
authors.lastname: Nakashima
authors.forename: Y
authors.initials: Y
authors.lastname: Nozawa
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D020730:Guanine Nucleotide Dissociation Inhibitors; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D006706:Homeostasis; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D015473:Leukemia, Promyelocytic, Acute; D008301:Maleimides / Q000494:pharmacology; D010739:Phospholipase D / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured; D062948:rho Guanine Nucleotide Dissociation Inhibitor alpha; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; D020742:rhoA GTP-Binding Protein
chemical_list: C569638:ARHGDIA protein, human; D020730:Guanine Nucleotide Dissociation Inhibitors; D007211:Indoles; D007527:Isoenzymes; D008301:Maleimides; D062948:rho Guanine Nucleotide Dissociation Inhibitor alpha; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; C071924:Ro 31-8425; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D011493:Protein Kinase C; D010739:Phospholipase D; D019204:GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein