B-cell activation by wild type and mutant Ig-beta cytoplasmic domains.
In B lymphocytes, the cytoplasmic domains of the membrane immunoglobulin-associated heterodimeric Ig-alpha and Ig-beta proteins link membrane immunoglobulin to intracellular signalling molecules. We constructed chimeric genes encoding the extracellular and transmembrane domain of human CD8 alpha and the cytoplasmic domain of Ig-alpha or Ig-beta and examined the ability of the chimeric proteins to induce signalling in the murine B-cell lymphoma A20. Crosslinking of CD8/Ig-alpha or CD8/Ig-beta induced both calcium mobilization and protein tyrosine phosphorylation, although induction by CD8/Ig-alpha was somewhat stronger. We also carried out mutagenesis of residues within the "Reth" motif of the CD8/Ig-beta cytoplasmic domain and determined the effects of these mutations on signalling in the murine B-cell hybridoma LK 35.2. Mutants in which alanine was substituted for glutamine 202, threonine 205, and isoleucine 209 retained the ability to induce protein tyrosine phosphorylation and calcium mobilization. In contrast, substitution of alanine for leucine 198 abrogated these responses, suggesting a critical role for this residue in interaction with cytoplasmic signalling proteins.
Taddie
J A
JA
Molecular Biology and Virology Laboratory, Salk Institute, San Diego, CA 92186.
Hurley
T R
TR
Sefton
B M
BM
D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D016827:CD8 Antigens / Q000235:genetics; D002118:Calcium / Q000378:metabolism; D017931:DNA Primers; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D016393:Lymphoma, B-Cell / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011947:Receptors, Antigen, B-Cell / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured
D016827:CD8 Antigens; D017931:DNA Primers; D011947:Receptors, Antigen, B-Cell; D011993:Recombinant Fusion Proteins; D002118:Calcium