B-Cell Activation by Wild Type and Mutant Ig-β Cytoplasmic Domains

  • John A. Taddie
  • Tamara R. Hurley
  • Bartholomew M. Sefton
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 365)

Abstract

In B lymphocytes, the cytoplasmic domains of the membrane immunoglobulin-associated heterodimeric Ig-α and Ig-β proteins link membrane immunoglobulin to intracellular signalling molecules. We constructed chimeric genes encoding the extracellular and transmembrane domain of human CD8α and the cytoplasmic domain of Ig-α or Ig-β and examined the ability of the chimeric proteins to induce signalling in the murine B-cell lymphoma A20. Crosslinking of CD8/Ig-α or CD8/Ig-β induced both calcium mobilization and protein tyrosine phosphorylation, although induction by CD8/Ig-α was somewhat stronger. We also carried out mutagenesis of residues within the “Reth” motif of the CD8/Ig-β 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.

Keywords

Cytoplasmic Domain Antigen Receptor Chimeric Protein Vesicular Stomatitis Virus Bovine Leukemia Virus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    J. Hombach, T. Tsubata, L. Leclercq, H. Stappert and M. Reth, Molecular components of the B-cell antigen receptor complex of the IgM class, Nature 343:760 (1990).PubMedCrossRefGoogle Scholar
  2. 2.
    J. Hombach, F. Lottspeich and M. Reth, Identification of the genes encoding the IgMα and Igβ components of the IgM antigen receptor complex by amino terminal sequencing, Eur. J. Immunol. 20:2795 (1990).PubMedCrossRefGoogle Scholar
  3. 3.
    K.S. Campbell, EJ. Hager, R.J. Friedrich and J.C. Cambier, IgM antigen receptor complex contains phosphoprotein products of B29 and mb-1 genes, Proc. Natl. Acad. Sci. USA 88:3982 (1991).PubMedCrossRefGoogle Scholar
  4. 4.
    A.R. Venkitaraman, G.T. Williams, P. Dariavach and M.S. Neuberger, The B-cell antigen receptor of the five immunoglobulin classes, Nature 352:777 (1991).PubMedCrossRefGoogle Scholar
  5. 5.
    M. Reth, Antigen receptors on B lymphocytes, Annu. Rev. Immunol. 10:97 (1992).PubMedCrossRefGoogle Scholar
  6. 6.
    M.-A. Campbell and B.M. Sefton, Protein tyrosine phosphorylation is induced in murine B lymphocytes in response to stimulation of anti-immunoglobulin, EMBO J. 9:2125 (1990).PubMedGoogle Scholar
  7. 7.
    M.R. Gold, D. Law and A.L. DeFranco, Stimulation of protein tyrosine phosphorylation by the B lymphocyte antigen receptor, Nature 345:810 (1990).PubMedCrossRefGoogle Scholar
  8. 8.
    M. Sanchez, Z. Misulovin, A.L. Burkhardt, S. Mahajan, T. Costa, R. Franke, J.B. Bolen and M. Nussenzweig, Signal transduction by immunoglobulin is mediated through Iga and Igβ, J. Exp. Med. 178:1049(1993).PubMedCrossRefGoogle Scholar
  9. 9.
    S.A. Grupp, K. Campbell, R.N. Mitchell, J.C. Cambier and A.K. Abbas, Signaling-defective mutants of the B lymphocyte antigen receptor fail to associate with Ig-α and Ig-β/γ, J. Biol. Chem. 268:25776 (1993).PubMedGoogle Scholar
  10. 10.
    K.-M. Kim, G. Alber, P. Weiser and M. Reth, Differential signalling through the Ig-α and Ig-β components of the B cell antigen receptor, Eur. J. Immunol. 23:911 (1993).PubMedCrossRefGoogle Scholar
  11. 11.
    D.A. Law, V.W.F. Chan, S.K. Datta and A.L. DeFranco, B-cell antigen receptor motifs have redundant signalling capabilities and bind the tyrosine kinases PTK72, Lyn, and Fyn, Current Biology 3:645 (1993).PubMedCrossRefGoogle Scholar
  12. 12.
    G.T. Williams, C.J.G. Peaker, K.J. Patel and M.S. Neuberger, The α/β sheath and its cytoplasmic tyrosine are required for signaling by the B-cell antigen receptor but not for capping or for serine/threonine recruitment, Proc. Natl. Acad. Sci. USA 91:474 (1994).PubMedCrossRefGoogle Scholar
  13. 13.
    J.A. Taddie, T.R. Hurley, B.S. Hardwick and B.M. Sefton, Activation of B and T cells by the cytoplasmic domains of the B-cell antigen receptor proteins Ig-α and Ig-β, J. Biol. Chem. In Press (1994).Google Scholar
  14. 14.
    M. Reth, Antigen receptor tail clue, Nature 338:383(1989).PubMedCrossRefGoogle Scholar
  15. 15.
    J.C. Cambier, Signal transduction by T-and B-cell antigen receptors: converging structures and concepts, Curr. Opin. Immunol. 4:257 (1992).PubMedCrossRefGoogle Scholar
  16. 16.
    G. Alber, K.-M. Kim, P. Weiser, C. Riesterer, R. Carsetti and M. Reth, Molecular mimicry of the antigen receptor signalling motif by transmembrane proteins of the Epstein-Barr virus and the bovine leukemia virus, Curr. Biol. 3:333 (1993).PubMedCrossRefGoogle Scholar
  17. 17.
    P. Beaufils, D. Choquet, R.Z. Mamoun and B. Malissen, The (YXXL/I)2 signalling motif found in the cytoplasmic segments of the bovine leukaemia virus envelope protein and Epstein-Barr virus latent membrane protein 2A can elicit early and late lymphocyte activation events, EMBO J. 12:5105 (1993).PubMedGoogle Scholar
  18. 18.
    F. Letourneur and R.D. Klausner, Activation of T cells by a tyrosine kinase activation domain in the cytoplasmic tail of CD3 ε, Science 255:79 (1992).PubMedCrossRefGoogle Scholar
  19. 19.
    C. Romeo, M. Amiot and B. Seed, Sequence requirements for induction of cytolysis by the T cell antigen/Fc receptor ζ chain, Cell 68:889 (1992).PubMedCrossRefGoogle Scholar
  20. 20.
    B.A. Irving, A.C. Chan and A. Weiss, Functional characterization of a signal transducing motif present in the T cell antigen receptor ζ chain, J. Exp. Med. 177:1093 (1993).PubMedCrossRefGoogle Scholar
  21. 21.
    K.J. Kim, C. Kanellopoulos-Langevin, R.M. Merwin, D.H. Sachs and R. Asofsky, Establishment and characterization of BALB/c lymphoma lines with B cell properties, J. Immunol. 122:549 (1979).PubMedGoogle Scholar
  22. 22.
    J. Kappler, J. White, D. Wegmann, E. Mustain and P. Marrack, Antigen presentation by Ia+ B cell hybridomas to H-2-restricted T cell hybridomas, Proc. Natl. Acad. Sci. USA 79:3604 (1982).PubMedCrossRefGoogle Scholar
  23. 23.
    G. Dennett, Cloned lines of natural killer cells, Nature 287:47 (1980).CrossRefGoogle Scholar
  24. 24.
    N.R. Landau and D.R. Littman, Packaging system for rapid production of murine leukemia virus vectors with variable tropism, J. Virol. 66: 5110 (1992).PubMedGoogle Scholar
  25. 25.
    T.R. Hurley, R. Hyman and B.M. Sefton, Differential effects of the CD45 tyrosine protein phosphatase on the tyrosine phosphorylation of the lck, fyn, and c-src tyrosine protein kinases, Mol. Cell. Biol. 13:1651 (1993).PubMedGoogle Scholar
  26. 26.
    T.R. Hurley and B.M. Sefton, Analysis of the activity and phosphorylation of the Ick protein in lymphoid cells, Oncogene 4:265 (1989).PubMedGoogle Scholar
  27. 27.
    D.J. Leahy, R. Axel and W.A. Hendrickson, Crystal structure of a soluble form of the human T cell coreceptor CD8 at 2.6 Å resolution, Cell 68: 1145 (1992).PubMedCrossRefGoogle Scholar
  28. 28.
    A.L. Burkhardt, T. Costa, Z. Misulovin, B. Stealy, J.B. Bolen and M.C. Nussenzweig, Igα and Igβ are functionally homologous to the signaling proteins of the T-cell receptor, Mol. Cell. Biol. 14:1095 (1994).PubMedGoogle Scholar
  29. 29.
    W. Kolanus, C. Romeo and B. Seed, Lineage-independent activation of immune system effector function by myeloid Fc receptors, EMBO J. 11:4861 (1992).PubMedGoogle Scholar
  30. 30.
    H. Flaswinkel and M. Reth, Dual role of the tyrosine activation motif of the Ig-α protein during signal transduction via the B cell antigen receptor, EMBO J. 13:83 (1994).PubMedGoogle Scholar
  31. 31.
    M.R. Clark, K.S. Campbell, A. Kazlauskas, S.A. Johnson, M. Hertz, T.A. Potter, C. Pleiman and J.C. Cambier, The B cell antigen receptor complex: association of Ig-α and Ig-β with distinct cytoplasmic effectors, Science 258:123 (1992).PubMedCrossRefGoogle Scholar
  32. 32.
    D.L. Law, M.R. Gold and A.L. DeFranco, Examination of B lymphoid cell lines for membrane immunoglobulin-stimulated tyrosine phosphorylation and src-family tyrosine kinase mRNA expression, Mol. Immunol. 29:917 (1992).PubMedCrossRefGoogle Scholar
  33. 33.
    Z. Songyang, S.E. Shoelson, M. Chaudhuri, G. Gish, T. Pawson, W.G. Haser, F. King, T. Roberts, S. Ratnofsky, R.J. Lechleider, B.G. Neel, R.B. Birge, J.E. Fajardo, M.M. Chou, H. Hanafusa, B. Schaffhausen and L.C. Cantley, SH2 domains recognize specific phosphopeptide sequences, Cell 72:767 (1993).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • John A. Taddie
    • 1
  • Tamara R. Hurley
    • 1
  • Bartholomew M. Sefton
    • 1
  1. 1.Molecular Biology and Virology LaboratoryThe Salk InstituteSan DiegoUSA

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