A requirement for Syk in the activation of the microtubule-associated protein kinase/phospholipase A₂ pathway by FcεR1 is not shared by a G protein-coupled receptor

Stimulation of the mast cell line, RBL-2H3, with antigen via the tetrameric (αβγ₂) immunoglobulin E receptor (FcεR1) leads to the activation of cytosolic phospholipase A₂ and the release of arachidonic acid. This pathway is dependent on the activation of the mitogen-activated protein (MAP) kinase. In this paper, we show that the MAP kinase/cytosolic phospholipase A₂ pathway is linked to FcεR1 via the cytosolic tyrosine kinase, Syk, and that the GDP/GTP exchange factor, Vav, might be one candidate for accomplishing this link. Cross-linking of transmembrane chimeras containing the FcεR1γ motif, which is known to activate Syk, results in the tyrosine phosphorylation of Vav, activation of MAP kinase, and release of arachidonic acid. Cross-linking of chimeras containing the FcεR1β motif does not cause these events. Furthermore, stimulation of these events by antigen is enhanced by transient overexpression of a wild-type form of Syk and blocked by overexpression of a dominant negative form of Syk. By contrast, stimulation via the transfected, G protein-coupled, muscarinic m1 receptor is not influenced by either form of Syk and does not result in tyrosine phosphorylation of Vav. These data establish unequivocally that the two types of receptor are independently linked to the MAP kinase/cytosolic phospholipase A₂ pathway and demonstrate the existence of the FcεR1-Syk- MAP kinase pathway.