Article

Tyrosine phosphorylation and complex formation of Cbl-b upon T cell receptor stimulation.

Division of Cell Biology, La Jolla Institute for Allergy and Immunology, San Diego, California 92121, USA.
Oncogene (impact factor: 6.37). 03/1999; 18(5):1147-56. DOI:10.1038/sj.onc.1202411 pp.1147-56
Source: PubMed

ABSTRACT Cbl-b, a mammalian homolog of Cbl, consists of an N-terminal region (Cbl-b-N) highly homologous to oncogenic v-Cbl, a Ring finger, and a C-terminal region containing multiple proline-rich stretches and potential tyrosine phosphorylation sites. In the present study, we demonstrate that upon engagement of the T cell receptor (TCR), endogenous Cbl-b becomes rapidly tyrosine-phosphorylated. In heterogeneous COS-1 cells, Cbl-b was phosphorylated on tyrosine residues by both Syk- (Syk/Zap-70) and Src- (Fyn/Lck) family kinases, with Syk kinase inducing the most prominent effect. Syk associates and phosphorylates Cbl-b in Jurkat T cells. A Tyr-316 Cbl-binding site in Syk was required for the association with and for the maximal tyrosine phosphorylation of Cbl-b. Mutation at a loss-of-function site (Gly-298) in Cbl-b-N disrupts its interaction with Syk. Cbl-b constitutively binds Grb2 and becomes associated with Crk-L upon TCR stimulation. The Grb2- and the Crk-L-binding regions were mapped to the C-terminus of Cbl-b. The Crk-L-binding sites were further determined to be Y655DVP and Y709KIP, with the latter being the primary binding site. Taken together, these results implicate that Cbl-b is involved in TCR-mediated intracellular signaling pathways.

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Keywords

C-terminal region
 
Cbl-b constitutively binds Grb2
 
Cbl-b-N
 
Cbl-b-N disrupts
 
Crk-L-binding regions
 
Crk-L-binding sites
 
endogenous Cbl-b
 
heterogeneous COS-1 cells
 
Jurkat T cells
 
maximal tyrosine phosphorylation
 
multiple proline-rich stretches
 
N-terminal region
 
phosphorylates Cbl-b
 
potential tyrosine phosphorylation sites
 
primary binding site
 
T cell receptor
 
TCR stimulation
 
TCR-mediated intracellular signaling pathways
 
Tyr-316 Cbl-binding site
 
tyrosine residues