Activation of the Cooh-Terminal Src Kinase (Csk) by Camp-Dependent Protein Kinase Inhibits Signaling through the T Cell Receptor

In T cells, cAMP-dependent protein kinase (PKA) type I colocalizes with the T cell receptor–CD3 complex (TCR/CD3) and inhibits T cell function via a previously unknown proximal target. Here we examine the mechanism for this PKA-mediated immunomodulation. cAMP treatment of Jurkat and normal T cells reduces Lck-mediated tyrosine phosphorylation of the TCR/CD3 ζ chain after T cell activation, and decreases Lck activity. Phosphorylation of residue Y505 in Lck by COOH-terminal Src kinase (Csk), which negatively regulates Lck, is essential for the inhibitory effect of cAMP on ζ chain phosphorylation. PKA phosphorylates Csk at S364 in vitro and in vivo leading to a two- to fourfold increase in Csk activity that is necessary for cAMP-mediated inhibition of TCR-induced interleukin 2 secretion. Both PKA type I and Csk are targeted to lipid rafts where proximal T cell activation occurs, and phosphorylation of raft-associated Lck by Csk is increased in cells treated with forskolin. We propose a mechanism whereby PKA through activation of Csk intersects signaling by Src kinases and inhibits T cell activation.

[1]  A. Wittinghofer,et al.  Epac is a Rap1 guanine-nucleotide-exchange factor directly activated by cyclic AMP , 1998, Nature.

[2]  E. Aandahl,et al.  Protein kinase A type I antagonist restores immune responses of T cells from HIV‐infected patients , 1998, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[3]  S. Cook,et al.  Inhibition by cAMP of Ras-dependent activation of Raf. , 1993, Science.

[4]  J. Partanen,et al.  cyl encodes a putative cytoplasmic tyrosine kinase lacking the conserved tyrosine autophosphorylation site (Y416src). , 1991, Oncogene.

[5]  T. Mustelin,et al.  Involvement of Src-homology-2-domain-containing protein-tyrosine phosphatase 2 in T cell activation. , 1996, European journal of biochemistry.

[6]  S. Taylor,et al.  Physiological inhibitors of the catalytic subunit of cAMP-dependent protein kinase: effect of MgATP on protein-protein interactions. , 1993, Biochemistry.

[7]  E. Aandahl,et al.  Additive effects of IL-2 and protein kinase A type I antagonist on function of T cells from HIV-infected patients on HAART. , 1999, AIDS.

[8]  L. Samelson,et al.  LAT palmitoylation: its essential role in membrane microdomain targeting and tyrosine phosphorylation during T cell activation. , 1998, Immunity.

[9]  Toshifumi Takao,et al.  Transmembrane phosphoprotein Cbp regulates the activities of Src-family tyrosine kinases , 2000, Nature.

[10]  K. Torgersen,et al.  Selective Activation of cAMP-dependent Protein Kinase Type I Inhibits Rat Natural Killer Cell Cytotoxicity* , 1997, The Journal of Biological Chemistry.

[11]  R. Klausner,et al.  T cell receptor tyrosine phosphorylation. Variable coupling for different activating ligands. , 1987, The Journal of biological chemistry.

[12]  Tomas Mustelin,et al.  Crosstalk between cAMP-dependent kinase and MAP kinase through a protein tyrosine phosphatase , 1999, Nature Cell Biology.

[13]  T. Vang,et al.  Kinetic properties of the C-terminal Src kinase, p50csk. , 1998, Biochimica et biophysica acta.

[14]  A. Gilman,et al.  Organization of G proteins and adenylyl cyclase at the plasma membrane. , 1997, Molecular biology of the cell.

[15]  J. Partanen,et al.  The human p50csk tyrosine kinase phosphorylates p56lck at Tyr‐505 and down regulates its catalytic activity. , 1992, The EMBO journal.

[16]  S. Rhee,et al.  Phospholipase C isozymes and modulation by cAMP-dependent protein kinase. , 1993, Advances in second messenger and phosphoprotein research.

[17]  L. Samelson,et al.  T cell antigen-receptor signal transduction. , 1999, Current opinion in immunology.

[18]  T. Yamamoto,et al.  CSK: a protein-tyrosine kinase involved in regulation of src family kinases. , 1991, The Journal of biological chemistry.

[19]  T. Mustelin,et al.  p56lck-independent activation and tyrosine phosphorylation of p72syk by T-cell antigen receptor/CD3 stimulation. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[20]  M. Lisanti,et al.  Regulation of cAMP-mediated Signal Transduction via Interaction of Caveolins with the Catalytic Subunit of Protein Kinase A* , 1999, The Journal of Biological Chemistry.

[21]  J. Bolen.  Nonreceptor tyrosine protein kinases. , 1993, Oncogene.

[22]  Karel Drbal,et al.  Phosphoprotein Associated with Glycosphingolipid-Enriched Microdomains (Pag), a Novel Ubiquitously Expressed Transmembrane Adaptor Protein, Binds the Protein Tyrosine Kinase Csk and Is Involved in Regulation of T Cell Activation , 2000, The Journal of experimental medicine.

[23]  M. Fournel,et al.  Negative regulation of T-cell receptor signalling by tyrosine protein kinase p50csk , 1993, Nature.

[24]  E. Aandahl,et al.  Increased activation of protein kinase A type I contributes to the T cell deficiency in common variable immunodeficiency. , 1999, Journal of immunology.

[25]  T. Mustelin.  T cell antigen receptor signaling: three families of tyrosine kinases and a phosphatase. , 1994, Immunity.

[26]  M. Crumpton,et al.  Comparative analysis of phosphotyrosyl polypeptides in normal and leukemic human T lymphocytes activated via CD3 or CD2. , 1993, Molecular immunology.

[27]  C. Rudd.  Adaptors and Molecular Scaffolds in Immune Cell Signaling , 1999, Cell.

[28]  Y. Takayama,et al.  Transmembrane Phosphoprotein Cbp Positively Regulates the Activity of the Carboxyl-terminal Src Kinase, Csk* , 2000, The Journal of Biological Chemistry.

[29]  T. Mustelin,et al.  Regulation of src family tyrosine kinases in lymphocytes. , 1993, Trends in biochemical sciences.

[30]  G. Crabtree,et al.  Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation , 1992, Nature.

[31]  R. Xavier,et al.  Membrane compartmentation is required for efficient T cell activation. , 1998, Immunity.

[32]  T. Mustelin,et al.  Rapid activation of the T-cell tyrosine protein kinase pp56lck by the CD45 phosphotyrosine phosphatase. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[33]  E. Smeland,et al.  Cyclic AMP‐dependent protein kinase (cAK) in human B cells: co‐localization of type I cAK (RIα2C2) with the antigen receptor during anti‐immunoglobulin‐induced B cell activation , 1996, European journal of immunology.

[34]  M. Colledge,et al.  AKAPs: from structure to function. , 1999, Trends in cell biology.

[35]  S. Døskeland,et al.  Cyclic AMP-dependent protein kinase type I mediates the inhibitory effects of 3',5'-cyclic adenosine monophosphate on cell replication in human T lymphocytes. , 1992, The Journal of biological chemistry.

[36]  J. Imboden,et al.  Molecular Cloning of the cDNA Encoding pp36, a Tyrosine-phosphorylated Adaptor Protein Selectively Expressed by T Cells and Natural Killer Cells , 1998, The Journal of experimental medicine.

[37]  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.

[38]  J. Frost,et al.  Regulation of the MAP kinase cascade. , 1994, Cellular & molecular biology research.

[39]  P. W. Janes,et al.  Aggregation of Lipid Rafts Accompanies Signaling via the T Cell Antigen Receptor , 1999, The Journal of cell biology.

[40]  H. Huitfeldt,et al.  Location of cAMP-dependent protein kinase type I with the TCR-CD3 complex. , 1994, Science.

[41]  D. Green,et al.  Modification of Phosphatidylinositol 3-Kinase SH2 Domain Binding Properties by Abl- or Lck-mediated Tyrosine Phosphorylation at Tyr-688* , 1998, The Journal of Biological Chemistry.