Asymmetrical phosphorylation and function of immunoreceptor tyrosine-based activation motif tyrosines in B cell antigen receptor signal transduction.
CD79a and CD79b function as transducers of B cell antigen receptor signals via a cytoplasmic sequence, termed the immunoreceptor tyrosine-based activation motif (ITAM). ITAMs contain two conserved tyrosines that may become phosphorylated upon receptor aggregation and bind distinct effectors by virtue of the distinct preference of phosphotyrosyl-containing sequences for SH2 domains. To explore the function of CD79a and CD79b ITAM tyrosines, we created membrane molecules composed of MHC class II I-Ak extracellular and transmembrane domains, and CD79a or CD79b cytoplasmic domains in which one or both of the ITAM tyrosines were mutated to phenylalanine. Functional analysis revealed that both ITAM tyrosines are required for ligand-induced Syk phosphorylation. However CD79a-ITAM and CD79b-ITAM tyrosine phosphorylations were asymmetrical, with >80% of phosphorylation occurring on the N-terminal tyrosine (Y-E-G-L). Thus, these findings suggest that following receptor ligation, only a minor proportion of phosphorylated ITAMs are doubly phosphorylated and thus can engage Syk. Only the N-terminal ITAM tyrosine of CD79a was required for ligand-mediated phosphorylation of the receptor and a subset of downstream substrates, including p62, p110, and Shc, and for Ca2+ mobilization. However, responses mediated through CD79b exhibited a greater dependence on the presence of both tyrosines. Neither tyrosine in CD79a or CD79b appeared absolutely essential for Src family kinase phosphorylation. These results indicate that phosphorylations of the tyrosines in CD79a and CD79b occur with very different stoichiometry, and the respective tyrosyl residues have distinct functions.
Pao
L I
LI
Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206, USA.
Famiglietti
S J
SJ
Cambier
J C
JC
D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D051925:CD79 Antigens; D000949:Histocompatibility Antigens Class II / Q000235:genetics; D006801:Humans; D008213:Lymphocyte Activation; D016393:Lymphoma, B-Cell; D016254:Mutagenesis, Insertional; D010649:Phenylalanine / Q000235:genetics; D010766:Phosphorylation; D017459:Receptors, Amino Acid / Q000235:genetics / Q000502:physiology*; D011947:Receptors, Antigen, B-Cell / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011971:Receptors, Immunologic / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000235:genetics / Q000502:physiology*
D015703:Antigens, CD; D051925:CD79 Antigens; C497883:CD79B protein, human; D000949:Histocompatibility Antigens Class II; D017459:Receptors, Amino Acid; D011947:Receptors, Antigen, B-Cell; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; C087659:tyrosine receptor; D014443:Tyrosine; D010649:Phenylalanine