Selective control of membrane ruffling and actin plaque assembly by the Rho GTPases Rac1 and CDC42 in FcepsilonRI-activated rat basophilic leukemia (RBL-2H3) cells.
Engagement of the high affinity IgE receptor (FcepsilonRI) in mast cells elicits a series of intracellular signalling events including cytoskeletal reorganization and granule exocytosis. To analyze the coupling of receptor activation to specific cytoskeletal responses, we expressed dominant negative mutant forms of the Rho GTPases CDC42 and Rac1 in rat RBL-2H3 tumor mast cells. We show here that dominant inhibition of CDC42 function decreases cell adhesion, interferes with Fc(epsilon)RI-induced actin plaque assembly and reduced the recruitment of vinculin at the cell-substratum interface, while the inhibitory Rac1 mutant abolishes Fc(epsilon)RI-mediated membrane ruffling. The expression of trans-dominant inhibitory forms of either CDC42 or Rac1 significantly inhibited antigen-induced degranulation. Altogether, our results demonstrate that CDC42 and Rac1 control distinct pathways downstream of FcepsilonRI engagement leading either to the induction of actin plaques, or to the production of membrane ruffles. These two pathways are critically involved during the degranulation response induced by Fc(epsilon)RI aggregation.
Guillemot
J C
JC
Centre d'Immunologie INSERM-CNRS de Marseille-Luminy, Marsaille, France.
Montcourrier
P
P
Vivier
E
E
Davoust
J
J
Chavrier
P
P
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D048429:Cell Size / Q000502:physiology; D003594:Cytoplasmic Granules / Q000502:physiology; D003599:Cytoskeleton / Q000737:chemistry / Q000201:enzymology; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D020690:GTPase-Activating Proteins; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015471:Leukemia, Basophilic, Acute; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017455:Receptors, IgE / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure; D016596:Vinculin / Q000032:analysis; D020764:cdc42 GTP-Binding Protein
D000199:Actins; D018797:Cell Cycle Proteins; D020690:GTPase-Activating Proteins; D011506:Proteins; D017455:Receptors, IgE; D016596:Vinculin; D012701:Serotonin; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D020764:cdc42 GTP-Binding Protein