RECRUITMENT AND PHOSPHORYLATION OF SH2-CONTAINING INOSITOL PHOSPHATASE AND SHE TO THE B-CELL FC-GAMMA IMMUNORECEPTOR TYROSINE-BASED INHIBITION MOTIF PEPTIDE MOTIF

Recently, we and others have demonstrated that negative signaling in B cells selectively induces the tyrosine phosphorylation of a novel ino sitol polyphosphate phosphatase, p145SHIP. In this study, we present d ata indicating that p145SHIP binds directly a phosphorylated motif, im munoreceptor tyrosine-based inhibition motif (ITIM), present in the cy toplasmic domain of Fc gamma RIIB1, Using recombinant SH2 domains, we show that binding is mediated via the Src homology region 2 (SH2)-cont aining inositol phosphatase (SHIP) SH2 domain, SHIP also bound to a ph osphopeptide derived from CD22, raising the possibility that SHIP cont ributes to negative signaling by this receptor as well as Fc gamma RII B1. The association of SHIP with the ITIM phosphopeptide was activatio n independent, while coassociation with She was activation dependent, Furthermore, experiments with Fc gamma RIIB1-deficient B cells demonst rated a genetic requirement for expression of Fc gamma RIIB1 in the in duction of SHIP phosphorylation and its interaction with Shc.. Based o n these results, we propose a model of negative signaling in which co- cross-linking of surface immunoglobulin and Fc gamma RIIB1 results in sequential tyrosine phosphorylation of the ITIM, recruitment and phosp horylation of p145SHIP, and subsequent binding of Shc.