In the present work, we explore the role of Syk in antigen-induced FcεRI endocytosis, investigating, in particular, whether Syk kinase activity controls the covalent modifications of Hrs, the main Ub-binding adapter implicated in sorting of engaged FcεRI complexes to lysosome for degradation [11, 18]. By siRNA knock down of Syk, we initially

support our previous evidence that in RBL-2H3 cells Syk is required for efficient internalization of engaged FcεRI [10]. Our results are in agreement with previous studies reporting that in macrophages Syk plays a major role in FcγR-mediated phagocytosis [33, 34] and in B cells EGFR inhibition is involved in both steady state and ligand-mediated BCR internalization [35]. However, our findings appear in contradiction with those of Bonnerot et al. [4] obtained using B lymphoma cells stably transfected with a chimeric receptor containing only FcεRI γ chain and of Kitaura et al. [36] showing that, in BMMCs, Syk has almost no effect on FcεRI endocytosis. A possible explanation for these contradictory findings is hat the contribution of Syk in regulating the internalization of γ chain containing receptors varies depending on receptor context (chimeric versus

endogenous multimeric receptor complex) and/or the source of cells used. Furthermore, Kitaura and coauthors [36] evaluated FcεRI internalization selleck chemicals only at 48 h after stimulation, leaving open the possibility that receptor internalization is affected at earlier time points. Our results, indeed, support a role for Syk in regulating mainly the early steps of antigen-induced FcεRI internalization: upon 30 min of stimulation almost 80% of the Syk knocked-down RBL-2H3

cells showed an impairment of FcεRI internalization, whereas upon 1 h of stimulation impaired FcεRI internalization was observed only in 50% of silenced cells analyzed. Thus, we would like to conclude that in mast cells Syk is required for a rapid and efficient antigen-induced FcεRI internalization, although 6-phosphogluconolactonase we cannot rule out that redundant mechanisms of receptor entry may also exist. Notably, in agreement with previous findings [4, 8], our results demonstrate a critical role for Syk in controlling the fate of internalized receptor complexes: Syk knockdown prevents the sorting of internalized receptors into lysosomes for degradation. This result was somewhat expected in light of our previous finding that c-Cbl-mediated ubiquitination of engaged FcεRI complexes is dependent on Syk kinase activity [17]. Indeed, by controlling receptor ubiquitination, Syk might indirectly affect receptor trafficking. In this respect, we have recently demonstrated a key role for the Ub pathway to ensure proper endocytic trafficking of engaged FcεRI complexes to the lysosomal compartment where degradation of the complexes can take place [11]. In addition, Syk kinase activity might control the action of molecular adapters directly implicated in the endocytic pathway.