Localized diacylglycerol-dependent stimulation of Ras and Rap1 during phagocytosis.
We describe a role for diacylglycerol in the activation of Ras and Rap1 at the phagosomal membrane. During phagocytosis, Ras density was similar on the surface and invaginating areas of the membrane, but activation was detectable only in the latter and in sealed phagosomes. Ras activation was associated with the recruitment of RasGRP3, a diacylglycerol-dependent Ras/Rap1 exchange factor. Recruitment to phagosomes of RasGRP3, which contains a C1 domain, parallels and appears to be due to the formation of diacylglycerol. Accordingly, Ras and Rap1 activation was precluded by antagonists of phospholipase C and of diacylglycerol binding. Ras is dispensable for phagocytosis but controls activation of extracellular signal-regulated kinase, which is partially impeded by diacylglycerol inhibitors. By contrast, cross-activation of complement receptors by stimulation of Fcgamma receptors requires Rap1 and involves diacylglycerol. We suggest a role for diacylglycerol-dependent exchange factors in the activation of Ras and Rap1, which govern distinct processes induced by Fcgamma receptor-mediated phagocytosis to enhance the innate immune response.
Botelho
Roberto J
RJ
Department of Chemistry and Biology, Ryerson University, Toronto, Ontario M5B 2K3, Canada.
Harrison
Rene E
RE
Stone
James C
JC
Hancock
John F
JF
Philips
Mark R
MR
Jongstra-Bilen
Jenny
J
Mason
David
D
Plumb
Jonathan
J
Gold
Michael R
MR
Grinstein
Sergio
S
D000818:Animals; D004075:Diglycerides / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D007113:Immunity, Innate; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D010587:Phagocytosis; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep; D015398:Signal Transduction; D020827:rap1 GTP-Binding Proteins / Q000378:metabolism*; D018631:ras Proteins / Q000378:metabolism*
D004075:Diglycerides; D007074:Immunoglobulin G; D020033:Protein Isoforms; D020827:rap1 GTP-Binding Proteins; D018631:ras Proteins
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