TY - JOUR
T1 - Activation of Syk tyrosine kinase is required for c-Cbl-mediated ubiquitination of FcεRI and Syk in RBL cells
AU - Paolini, Rossella
AU - Molfetta, Rosa
AU - Beitz, Laurie O.
AU - Zhang, Juan
AU - Scharenberg, Andrew M.
AU - Piccoli, Mario
AU - Frati, Luigi
AU - Siraganian, Reuben
AU - Santoni, Angela
PY - 2002/10/4
Y1 - 2002/10/4
N2 - Engagement of the high affinity receptor for IgE (FcεRI) on mast cells and basophils results in FcεRI β and γ subunits ubiquitination by an as yet undefined mechanism. Here we show that, upon FcεRI engagement on RBL-2H3 cells Syk undergoes ubiquitination and Syk kinase activity is required for its own ubiquitination and that of FcεRI β and γ chains. This requirement was demonstrated by overexpression of Syk wild-type or its kinase-dead mutant in RBL cells or using an Syk-deficient RBL-derived cell line transfected with wild-type or a kinase inactive form of Syk. We also identify c-Cbl as the E3 ligase responsible for both Syk and receptor ubiquitination. Furthermore, we demonstrate that Syk controls tyrosine phosphorylation of Syk-associated Cbl induced after receptor engagement. These data suggest a mutual regulation between Syk and Cbl activities. Finally, we show that a selective inhibitor of proteasome degradation induces persistence of tyrosine-phosphorylated receptor complexes, of activated Syk, and of FcεRI-triggered degranulation. Our results provide a molecular mechanism for down-regulation of engaged receptor complexes by targeting ubiquitinated FcεRI and activated Syk to the proteasome for degradation.
AB - Engagement of the high affinity receptor for IgE (FcεRI) on mast cells and basophils results in FcεRI β and γ subunits ubiquitination by an as yet undefined mechanism. Here we show that, upon FcεRI engagement on RBL-2H3 cells Syk undergoes ubiquitination and Syk kinase activity is required for its own ubiquitination and that of FcεRI β and γ chains. This requirement was demonstrated by overexpression of Syk wild-type or its kinase-dead mutant in RBL cells or using an Syk-deficient RBL-derived cell line transfected with wild-type or a kinase inactive form of Syk. We also identify c-Cbl as the E3 ligase responsible for both Syk and receptor ubiquitination. Furthermore, we demonstrate that Syk controls tyrosine phosphorylation of Syk-associated Cbl induced after receptor engagement. These data suggest a mutual regulation between Syk and Cbl activities. Finally, we show that a selective inhibitor of proteasome degradation induces persistence of tyrosine-phosphorylated receptor complexes, of activated Syk, and of FcεRI-triggered degranulation. Our results provide a molecular mechanism for down-regulation of engaged receptor complexes by targeting ubiquitinated FcεRI and activated Syk to the proteasome for degradation.
UR - http://www.scopus.com/inward/record.url?scp=0037020143&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037020143&partnerID=8YFLogxK
U2 - 10.1074/jbc.M204948200
DO - 10.1074/jbc.M204948200
M3 - Article
C2 - 12145291
AN - SCOPUS:0037020143
VL - 277
SP - 36940
EP - 36947
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 40
ER -