title: Co-expression with BCR induces activation of the FES tyrosine kinase and phosphorylation of specific N-terminal BCR tyrosine residues.
abstract: The human BCR gene encodes a protein with serine/threonine kinase activity and regulatory domains for the small G-proteins RAC and CDC42. Previous work in our laboratory has established that BCR is a substrate for c-FES, a non-receptor tyrosine kinase linked to myeloid growth and differentiation. Tyrosine phosphorylation led to the association of BCR with the RAS guanine nucleotide exchange complex GRB2-SOS in vivo via the GRB2 SH2 domain, linking BCR to RAS signaling (Maru, Y., Peters, K. L., Afar, D. E. H., Shibuya, M., Witte, O. N., and Smithgall, T. E. (1995) Mol. Cell. Biol. 15, 835-842). In the present study, we demonstrate that BCR Tyr-246 and at least one of the closely spaced tyrosine residues, Tyr-279, Tyr-283, and Tyr-289 (3Y cluster), are phosphorylated by FES both in vitro and in 32Pi-labeled cells. Mutagenesis of BCR Tyr-177 to Phe completely abolished FES-induced BCR binding to the GRB2 SH2 domain, identifying Tyr-177 as an additional phosphorylation site for FES. Co-expression of BCR and FES in human 293T cells stimulated the tyrosine autophosphorylation of FES. By contrast, tyrosine phosphorylation of BCR by FES suppressed BCR serine/threonine kinase activity toward the 14-3-3 protein and BCR substrate, BAP-1. These data show that tyrosine phosphorylation by FES affects the interaction of BCR with multiple signaling partners and suggest a general role for BCR in non-receptor protein-tyrosine kinase regulation and signal transduction.
authors.affiliation: Eppley Institute for Research in Cancer and Department of Pharmacology, University of Nebraska Medical Center, Omaha, Nebraska 68198-6805, USA. tsmithga@unmc.edu
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: T E
authors.initials: TE
authors.lastname: Smithgall
mesh_terms: D048948:14-3-3 Proteins; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D006801:Humans; D015513:Oncogene Proteins / Q000378:metabolism*; D010449:Peptide Mapping; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases; D011506:Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D051562:Proto-Oncogene Proteins c-bcr; D051578:Proto-Oncogene Proteins c-fes; D011994:Recombinant Proteins; D015398:Signal Transduction; D018411:Spodoptera; D014446:Tyrosine 3-Monooxygenase; D018909:src Homology Domains
chemical_list: D048948:14-3-3 Proteins; D015513:Oncogene Proteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D011994:Recombinant Proteins; D019000:Phosphotyrosine; D014446:Tyrosine 3-Monooxygenase; D011505:Protein-Tyrosine Kinases; C496645:FES protein, human; D051578:Proto-Oncogene Proteins c-fes; C496613:BCR protein, human; D051562:Proto-Oncogene Proteins c-bcr; D002265:Carboxylic Ester Hydrolases