title: Involvement of Rho family proteins in prostaglandin F2 alpha-induced phospholipase D activation in the osteoblast-like cell line MC3T3-E1.
abstract: To examine the role of Rho family proteins in prostaglandin F2 alpha (PGF2 alpha)-mediated phospholipase D (PLD) activation of osteoblast-like cell line MC3T3-E1 cells, we used Toxin-B from Clostridium difficile, which inhibits Rho family proteins by monoglucosylation. Pretreatment of [3H]myristic acid-labeled MC3T3-E1 cells with Toxin B induced rounding-up of the cells and inhibited the PGF2 alpha-induced PLD activation by 60%, but not the phospholipase C (PLC) activation. Cytochalasin D also induced rounding the cells, but showed a small inhibition in the PLD activation. Brefeldin A (BFA) had marginal inhibitory effect on the PGF2 alpha-induced PLD activation. In digitonin-permeabilized MC3T3-E1 cells, [3H]P But formation was stimulated by guanosine 5'-O-(3-thiotriphosphate) (GTP gamma S) or 4 beta-phorbol 12-myristate 13-acetate (PMA) in the presence of Ca2+ (1 microM) and ATP (1 mM), and phosphatidylinositol 4,5-bisphosphate (PIP2) was also required for its full PLD activation. Pretreatment of the digitonin-permeabilized MC3T3-E1 cells with Toxin B reduced the GTP gamma S- and PMA-stimulated PLD activities by 80% and 60%, respectively. On the other hand, C3 toxin which inhibits Rho by ADP-ribosylation, exerted a partial inhibitory effect on the GTP gamma S-stimulated PLD activity. These results suggest that Cdc42 as well as RhoA appear to be involved in the PLD activation mediated by PGF2 alpha and also that the PLD activation may be independent of actin cytoskeleton in MC3T3-E1 cells.
authors.affiliation: Department of Oral, and Maxillo-Facial Surgery, Gifu University School of Medicine, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kato
authors.forename: Y
authors.initials: Y
authors.lastname: Banno
authors.forename: T
authors.initials: T
authors.lastname: Dohjima
authors.forename: N
authors.initials: N
authors.lastname: Kato
authors.forename: K
authors.initials: K
authors.lastname: Watanabe
authors.forename: N
authors.initials: N
authors.lastname: Tatematsu
authors.forename: Y
authors.initials: Y
authors.lastname: Nozawa
mesh_terms: D036002:ADP Ribose Transferases / Q000494:pharmacology; D000199:Actins / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000494:pharmacology; D001905:Botulinum Toxins; D018797:Cell Cycle Proteins / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects; D004072:Digitonin / Q000494:pharmacology; D015237:Dinoprost / Q000378:metabolism / Q000494:pharmacology*; D004768:Enterotoxins / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000378:metabolism*; D020690:GTPase-Activating Proteins; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects / Q000378:metabolism*; D010739:Phospholipase D / Q000187:drug effects / Q000378:metabolism*; D011506:Proteins / Q000187:drug effects / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein
chemical_list: D000199:Actins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D018797:Cell Cycle Proteins; D004768:Enterotoxins; D020690:GTPase-Activating Proteins; D011506:Proteins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D015237:Dinoprost; D036002:ADP Ribose Transferases; C063007:exoenzyme C3, Clostridium botulinum; D010739:Phospholipase D; D001905:Botulinum Toxins; D019204:GTP-Binding Proteins; D020764:cdc42 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein; D004072:Digitonin; D013755:Tetradecanoylphorbol Acetate