Receptor Signaling and Syk in the Initiation of B-Cell Antigen Nonredundant Roles of Src-Family Kinases

When a BCR on a mature B cell is engaged by its ligand, the cell becomes activated, and the Ab-mediated immune response can be triggered. The initiation of BCR signaling is orchestrated by kinases of the Src and Syk families. However, the proximal BCR-induced phosphorylation remains incompletely understood. According to a model of sequential activation of kinases, Syk acts downstream of Src family kinases (SFKs). In addition, signaling independent of SFKs and initiated by Syk has been proposed. Both hypotheses lack sufficient evidence from relevant B cell models, mainly because of the redundancy of Src family members and the importance of BCR signaling for B cell development. We addressed this issue by analyzing controlled BCR triggering ex vivo on primary murine B cells and on murine and chicken B cell lines. Chemical and Csk-based genetic inhibitor treatments revealed that SFKs are required for signal initiation and Syk activation. In addition, ligand and anti-BCR Ab–induced signaling differ in their sensitivity to the inhibition of SFKs. The Journal of Immunology , 2013, 190: 000–000. earliest events of immunoreceptor signaling include catalyzed kinases the Src and Syk families. Src family kinases (SFKs) are cytoplasmic proteins that are associated with the plasma membrane via lipid at their N termini At members and cells the regulation of SFK activity on the phosphorylation of two key tyrosine residues. Phosphorylation of the C-terminal tyrosine (Y527 for chicken Src) is by Csk and counteracted by phosphatases CD45 and in

[1]  F. Rieux-Laucat,et al.  Primary T-cell immunodeficiency with immunodysregulation caused by autosomal recessive LCK deficiency. , 2012, The Journal of allergy and clinical immunology.

[2]  Boris N. Kholodenko,et al.  Emergence of bimodal cell population responses from the interplay between analog single-cell signaling and protein expression noise , 2012, BMC Systems Biology.

[3]  T. Kurosaki.  Regulation of BCR signaling. , 2011, Molecular immunology.

[4]  T. Kalina,et al.  Regulation of Src Family Kinases Involved in T Cell Receptor Signaling by Protein-tyrosine Phosphatase CD148* , 2011, The Journal of Biological Chemistry.

[5]  A. Michie,et al.  Dasatinib inhibits B cell receptor signalling in chronic lymphocytic leukaemia but novel combination approaches are required to overcome additional pro‐survival microenvironmental signals , 2011, British journal of haematology.

[6]  S. Pata,et al.  The effects of membrane compartmentalization of csk on TCR signaling. , 2011, Biochimica et biophysica acta.

[7]  J. M. Bradshaw,et al.  The Src, Syk, and Tec family kinases: distinct types of molecular switches. , 2010, Cellular signalling.

[8]  Mauro Riccaboni,et al.  Spleen tyrosine kinases: biology, therapeutic targets and drugs. , 2010, Drug discovery today.

[9]  Omer Dushek,et al.  Constitutively Active Lck Kinase in T Cells Drives Antigen Receptor Signal Transduction , 2010, Immunity.

[10]  Victor L. J. Tybulewicz,et al.  The SYK tyrosine kinase: a crucial player in diverse biological functions , 2010, Nature Reviews Immunology.

[11]  J. Leonard,et al.  Activities of SYK and PLCγ2 Predict Apoptotic Response of CLL Cells to SRC Tyrosine Kinase Inhibitor Dasatinib , 2010, Clinical Cancer Research.

[12]  M. Poderycki,et al.  A Minor Catalytic Activity of Src Family Kinases Is Sufficient for Maximal Activation of Mast Cells via the High-Affinity IgE Receptor , 2009, The Journal of Immunology.

[13]  Michael Reth,et al.  Autoinhibition and adapter function of Syk , 2009, Immunological reviews.

[14]  T. Kurosaki,et al.  Tyrosine kinases and their substrates in B lymphocytes , 2009, Immunological reviews.

[15]  Jayajit Das,et al.  Digital Signaling and Hysteresis Characterize Ras Activation in Lymphoid Cells , 2009, Cell.

[16]  N. K. Williams,et al.  Crystal Structures of the Lyn Protein Tyrosine Kinase Domain in Its Apo- and Inhibitor-bound State* , 2009, Journal of Biological Chemistry.

[17]  Emily K. Tsang,et al.  Molecular Mechanism of the Syk Activation Switch* , 2008, Journal of Biological Chemistry.

[18]  A. Weiss,et al.  Structurally distinct phosphatases CD45 and CD148 both regulate B cell and macrophage immunoreceptor signaling. , 2008, Immunity.

[19]  T. Kawakami,et al.  Positive and Negative Regulation of Mast Cell Activation by Lyn via the FcεRI1 , 2005, The Journal of Immunology.

[20]  A. Weiss,et al.  Intramolecular Regulatory Switch in ZAP-70: Analogy with Receptor Tyrosine Kinases , 2005, Molecular and Cellular Biology.

[21]  S. Gauld,et al.  Src-family kinases in B-cell development and signaling , 2004, Oncogene.

[22]  C. Lowell.  Src-family kinases: rheostats of immune cell signaling. , 2004, Molecular immunology.

[23]  K. Nakashima,et al.  The Orally Available Spleen Tyrosine Kinase Inhibitor 2-[7-(3,4-Dimethoxyphenyl)-imidazo[1,2-c]pyrimidin-5-ylamino]nicotinamide Dihydrochloride (BAY 61-3606) Blocks Antigen-Induced Airway Inflammation in Rodents , 2003, Journal of Pharmacology and Experimental Therapeutics.

[24]  P. Cohen,et al.  The specificities of protein kinase inhibitors: an update. , 2003, The Biochemical journal.

[25]  A. Tarakhovsky,et al.  Essential role of Src-family protein tyrosine kinases in NF-κB activation during B cell development , 2003, Nature Immunology.

[26]  C. Quilici,et al.  Sustained Activation of Lyn Tyrosine Kinase In Vivo Leads to Autoimmunity , 2002, The Journal of experimental medicine.

[27]  M. Reth,et al.  Amplification of B cell antigen receptor signaling by a Syk/ITAM positive feedback loop. , 2002, Molecular cell.

[28]  J. D. Dal Porto,et al.  B Cell Antigen Receptor Signaling: Roles in Cell Development and Disease , 2002, Science.

[29]  Jonathan C. Poe,et al.  A CD19-Dependent Signaling Pathway Regulates Autoimmunity in Lyn-Deficient Mice1 , 2001, The Journal of Immunology.

[30]  M. Fujimoto,et al.  CD19 regulates Src family protein tyrosine kinase activation in B lymphocytes through processive amplification. , 2000, Immunity.

[31]  R. Burton,et al.  Substrate Recognition by the Lyn Protein-tyrosine Kinase , 2000, The Journal of Biological Chemistry.

[32]  L. Toledo,et al.  Structural analysis of the lymphocyte-specific kinase Lck in complex with non-selective and Src family selective kinase inhibitors. , 2000, Structure.

[33]  J. D. Di Santo,et al.  Tyrosine kinase SYK: essential functions for immunoreceptor signalling. , 2000, Immunology today.

[34]  Juan Zhang,et al.  CD45 Is Essential for FcεRI Signaling by ZAP70, But Not Syk, in Syk-Negative Mast Cells , 1999, The Journal of Immunology.

[35]  J. Kuriyan,et al.  Crystal structure of Hck in complex with a Src family-selective tyrosine kinase inhibitor. , 1999, Molecular cell.

[36]  R. Geahlen,et al.  Syk- and Lyn-dependent phosphorylation of Syk on multiple tyrosines following B cell activation includes a site that negatively regulates signaling. , 1998, Journal of immunology.

[37]  Teruaki Kimura,et al.  Mutations in the activation loop tyrosines of protein tyrosine kinase Syk abrogate intracellular signaling but not kinase activity. , 1998, Journal of immunology.

[38]  K. Horikawa,et al.  A Double-Edged Kinase Lyn: A Positive and Negative Regulator for Antigen Receptor–mediated Signals , 1998, The Journal of experimental medicine.

[39]  J. Cambier,et al.  Asymmetrical phosphorylation and function of immunoreceptor tyrosine-based activation motif tyrosines in B cell antigen receptor signal transduction. , 1998, Journal of immunology.

[40]  Philippe Soriano,et al.  Characterization of the B lymphocyte populations in Lyn-deficient mice and the role of Lyn in signal initiation and down-regulation. , 1997, Immunity.

[41]  T. Kurosaki,et al.  Molecular mechanisms in B cell antigen receptor signaling. , 1997, Current opinion in immunology.

[42]  T. Mustelin,et al.  Role of Tyr518 and Tyr519 in the regulation of catalytic activity and substrate phosphorylation by Syk protein-tyrosine kinase. , 1997, European journal of biochemistry.

[43]  J. Cambier,et al.  Syk, but not Lyn, recruitment to B cell antigen receptor and activation following stimulation of CD45- B cells. , 1997, Journal of immunology.

[44]  J. Kinet,et al.  syk kinase activation by a src kinase-initiated activation loop phosphorylation chain reaction. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[45]  K. Rajewsky,et al.  B cell development under the condition of allelic inclusion. , 1997, Immunity.

[46]  J. Brugge,et al.  Protein tyrosine kinases Syk and ZAP-70 display distinct requirements for Src family kinases in immune response receptor signal transduction. , 1997, Journal of immunology.

[47]  R. Geahlen,et al.  Identification of the major sites of autophosphorylation of the murine protein-tyrosine kinase Syk. , 1997, Biochimica et biophysica acta.

[48]  A. Weiss,et al.  The Syk protein tyrosine kinase can function independently of CD45 or Lck in T cell antigen receptor signaling. , 1996, The EMBO journal.

[49]  A. Veillette,et al.  Differential Intrinsic Enzymatic Activity of Syk and Zap-70 Protein-tyrosine Kinases* , 1996, The Journal of Biological Chemistry.

[50]  J. Hanke,et al.  Discovery of a Novel, Potent, and Src Family-selective Tyrosine Kinase Inhibitor , 1996, The Journal of Biological Chemistry.

[51]  K. Sada,et al.  Role of the Syk autophosphorylation site and SH2 domains in B cell antigen receptor signaling , 1995, The Journal of experimental medicine.

[52]  J. Cambier,et al.  Phosphorylated immunoreceptor signaling motifs (ITAMs) exhibit unique abilities to bind and activate Lyn and Syk tyrosine kinases. , 1995, Journal of immunology.

[53]  T. Kurosaki,et al.  The catalytic activity of Src‐family tyrosine kinase is required for B cell antigen receptor signaling , 1995, FEBS letters.

[54]  S. Stacker,et al.  Multiple defects in the immune system of Lyn-deficient mice, culminating in autoimmune disease , 1995, Cell.

[55]  M. Nussenzweig,et al.  The role of Ig beta in precursor B cell transition and allelic exclusion. , 1995, Science.

[56]  T. Yamamoto,et al.  Syk activation by the Src-family tyrosine kinase in the B cell receptor signaling , 1994, The Journal of experimental medicine.

[57]  A. Hata,et al.  Tyrosine kinases Lyn and Syk regulate B cell receptor‐coupled Ca2+ mobilization through distinct pathways. , 1994, The EMBO journal.

[58]  A. Weiss,et al.  Sequential interactions of the TCR with two distinct cytoplasmic tyrosine kinases. , 1994, Science.

[59]  Dan R. Littman,et al.  Signal transduction by lymphocyte antigen receptors , 1994, Cell.

[60]  M. Reth,et al.  Dual role of the tyrosine activation motif of the Ig‐alpha protein during signal transduction via the B cell antigen receptor. , 1994, The EMBO journal.

[61]  A. Shaw,et al.  Palmitylation of an amino-terminal cysteine motif of protein tyrosine kinases p56lck and p59fyn mediates interaction with glycosyl-phosphatidylinositol-anchored proteins , 1993, Molecular and cellular biology.

[62]  S. Baylor,et al.  Use of fura red as an intracellular calcium indicator in frog skeletal muscle fibers. , 1993, Biophysical journal.

[63]  A. Weiss,et al.  CD45 specifically modulates binding of Lck to a phosphopeptide encompassing the negative regulatory tyrosine of Lck. , 1993, The EMBO journal.

[64]  Arthur Weiss,et al.  Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor , 1992, Cell.

[65]  A. Wakeham,et al.  Profound block in thymocyte development in mice lacking p56lck , 1992, Nature.

[66]  R. Kelly,et al.  Tyrosine phosphorylation of components of the B-cell antigen receptors following receptor crosslinking. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[67]  M. Reth,et al.  Membrane IgM and IgD molecules fail to transduce Ca2+ mobilizing signals when expressed on differentiated B lineage cells. , 1990, Journal of immunology.

[68]  M. Okada,et al.  A protein tyrosine kinase involved in regulation of pp60c-src function. , 1989, The Journal of biological chemistry.

[69]  D. Allen,et al.  Antibody engineering for the analysis of affinity maturation of an anti‐hapten response. , 1988, The EMBO journal.

[70]  T. Roberts,et al.  Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src , 1987, Cell.

[71]  A. Cumano,et al.  Clonal recruitment and somatic mutation in the generation of immunological memory to the hapten NP. , 1986, The EMBO journal.

[72]  F. Cross,et al.  A short sequence in the p60src N terminus is required for p60src myristylation and membrane association and for cell transformation , 1984, Molecular and cellular biology.

[73]  E. Ingley,et al.  Src family kinases: regulation of their activities, levels and identification of new pathways. , 2008, Biochimica et biophysica acta.

[74]  M. Hibbs,et al.  Lyn tyrosine kinase: accentuating the positive and the negative. , 2005, Immunity.

[75]  T. Kawakami,et al.  Positive and negative regulation of mast cell activation by Lyn via the FcepsilonRI. , 2005, Journal of immunology.

[76]  Wei Wang,et al.  Effect of autophosphorylation on the catalytic and regulatory properties of protein tyrosine kinase Src. , 2002, Archives of biochemistry and biophysics.

[77]  J. Zhang,et al.  CD45 is essential for Fc epsilon RI signaling by ZAP70, but not Syk, in Syk-negative mast cells. , 1999, Journal of immunology.