In RBL-2H3 rat tumor mast cells, cross-linking the high affinity IgE receptor, FcR1, activates the protein-tyrosine kinases Lyn and Syk and initiates a series of responses including protein-tyrosine phosphorylation, inositol 1,4,5-trisphosphate synthesis, Ca mobilization, secretion, membrane ruffling, and actin plaque assembly. The development of chimeric receptors containing cytoplasmic domains of individual subunits of the heterotrimeric () FcR1 has simplified analyses of early signaling events in RBL-2H3 cells. Here, RBL-2H3 cells were transfected with cDNAs encoding the extracellular and transmembrane domains of the interleukin-2 receptor subunit (the Tac antigen) joined to the C-terminal cytoplasmic domains of the FcR1 and subunits (TT and TT). Both sequences contain tyrosine activation motifs implicated in antigen receptor signal transduction. TT and TT are expressed independently of the native FcR1, as demonstrated by the ability of Tac cross-linking agents to trigger the clustering and internalization through coated pits of both chimeric receptors without co-clustering the FcR1. A full range of signaling activities is induced by TT cross-linking; the TT-induced responses are slower and, except for Lyn activation, smaller than the FcR1-induced responses. In striking contrast, TT cross-linking elicits no tyrosine phosphorylation or signaling responses, it impairs basal activities measured in secretion and anti-PY (anti-phosphotyrosine antibody) immune complex kinase assays, and it antagonizes FcR1-induced Lyn and Syk activation, protein-tyrosine phosphorylation, and signaling responses. We hypothesize that the isolated subunit binds a specific kinase or coupling protein(s) required for signaling activity, sequestering it from the signal-transducing subunit. Binding the same kinase or coupling protein to the subunit of the intact FcR1 may serve instead to present it to the adjacent subunit, resulting in enhanced kinase activation and signaling responses.

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