Reconstitution of high affinity IgE receptor-mediated secretion by transfecting protein tyrosine kinase pp125(FAK)

To study the role of the focal adhesion tyrosine kinase (FAK) in receptor-mediated secretion, we transfected FAK cDNA into a variant (3B6) of the RBL-2H3 mast cell line. This 3B6 cell line expressed low levels of FAK and was defective in high affinity IgE receptor (Fc epsilon RI) but not Ca2+ ionophore-mediated secretion. Fc epsilon RI-mediated secretion was reconstituted after transfection of wild-type FAK. Histamine release was also enhanced by the stable expression of two mutants of FAK: a kinase-inactive form in which the ATP binding site Lys-454 was replaced by Arg or a mutant in which the autophosphorylation site Tyr-397 was replaced by Phe. Therefore, the catalytic activity and the autophosphorylation site of FAK are not essential for secretion. Fc epsilon RI aggregation increased. the tyrosine phosphorylation of both mutants of FAK to the same extent as wild-type FAK. Therefore, tyrosine kinases activated by Fc epsilon RI aggregation are phosphorylating FAK and some of these phosphorylation sites are other than Tyr-397. These results strongly suggest that FAK plays a role in Fc epsilon RI-induced secretion by functioning as an adapter or linker molecule.