Amplification of receptor signalling by Ca2+ entry‐mediated translocation and activation of PLCγ2 in B lymphocytes

In non‐excitable cells, receptor‐activated Ca2+ signalling comprises initial transient responses followed by a Ca2+ entry‐dependent sustained and/or oscillatory phase. Here, we describe the molecular mechanism underlying the second phase linked to signal amplification. An in vivo inositol 1,4,5‐trisphosphate (IP3) sensor revealed that in B lymphocytes, receptor‐activated and store‐operated Ca2+ entry greatly enhanced IP3 production, which terminated in phospholipase Cγ2 (PLCγ2)‐deficient cells. Association between receptor‐activated TRPC3 Ca2+ channels and PLCγ2, which cooperate in potentiating Ca2+ responses, was demonstrated by co‐immunoprecipitation. PLCγ2‐deficient cells displayed diminished Ca2+ entry‐induced Ca2+ responses. However, this defect was canceled by suppressing IP3‐induced Ca2+ release, implying that IP3 and IP3 receptors mediate the second Ca2+ phase. Furthermore, confocal visualization of PLCγ2 mutants demonstrated that Ca2+ entry evoked a C2 domain‐mediated PLCγ2 translocation towards the plasma membrane in a lipase‐independent manner to activate PLCγ2. Strikingly, Ca2+ entry‐activated PLCγ2 maintained Ca2+ oscillation and extracellular signal‐regulated kinase activation downstream of protein kinase C. We suggest that coupling of Ca2+ entry with PLCγ2 translocation and activation controls the amplification and co‐ordination of receptor signalling.

[1]  M. Berridge.  Inositol trisphosphate and calcium signalling , 1993, Nature.

[2]  Robin F Irvine,et al.  Nuclear Lipid Signaling , 2000, Science's STKE.

[3]  S. Pentyala,et al.  Structure, function, and control of phosphoinositide-specific phospholipase C. , 2000, Physiological reviews.

[4]  J. D. Clark,et al.  A novel arachidonic acid-selective cytosolic PLA2 contains a Ca2+-dependent translocation domain with homology to PKC and GAP , 1991, Cell.

[5]  H. Kung,et al.  Phospholipase C-gamma 1 can induce DNA synthesis by a mechanism independent of its lipase activity. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R Y Tsien,et al.  Calcium channels, stores, and oscillations. , 1990, Annual review of cell biology.

[7]  R. Hurst,et al.  trp, a Novel Mammalian Gene Family Essential for Agonist-Activated Capacitative Ca2+ Entry , 1996, Cell.

[8]  S. Ryu,et al.  Direct interaction of SOS1 Ras exchange protein with the SH3 domain of phospholipase C-gamma1. , 2000, Biochemistry.

[9]  Roger L. Williams,et al.  Crystal structure of a mammalian phosphoinositide-specific phospholipase Cδ , 1996, Nature.

[10]  R. Penner,et al.  The Store-Operated Calcium Current ICRAC: Nonlinear Activation by InsP3 and Dissociation from Calcium Release , 1997, Cell.

[11]  K. Mikoshiba,et al.  Block of Ca2+ wave and Ca2+ oscillation by antibody to the inositol 1,4,5-trisphosphate receptor in fertilized hamster eggs. , 1992, Science.

[12]  A. Benedetti,et al.  Diacylglycerol activates the influx of extracellular cations in T-lymphocytes independently of intracellular calcium-store depletion and possibly involving endogenous TRP6 gene products. , 2002, The Biochemical journal.

[13]  P. Delmas,et al.  Signaling Microdomains Define the Specificity of Receptor-Mediated InsP3 Pathways in Neurons , 2002, Neuron.

[14]  J. Putney,et al.  Signaling Mechanism for Receptor-activated Canonical Transient Receptor Potential 3 (TRPC3) Channels* , 2003, The Journal of Biological Chemistry.

[15]  D. Clapham,et al.  The TRPM7 channel is inactivated by PIP2 hydrolysis , 2002, Nature Cell Biology.

[16]  F. A. Lai,et al.  PLC zeta: a sperm-specific trigger of Ca(2+) oscillations in eggs and embryo development. , 2002, Development.

[17]  T. Gudermann,et al.  Diffusion‐limited translocation mechanism of protein kinase C isotypes , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[18]  D. Clapham,et al.  Calcium signaling , 1995, Cell.

[19]  O. Petersen,et al.  The endoplasmic reticulum as an integrating signalling organelle: from neuronal signalling to neuronal death. , 2002, European journal of pharmacology.

[20]  L. Cantley,et al.  Activation of Phospholipase C-γ by Phosphatidylinositol 3,4,5-Trisphosphate* , 1998, The Journal of Biological Chemistry.

[21]  T. Kurosaki,et al.  Genetic evidence for involvement of type 1, type 2 and type 3 inositol 1,4,5‐trisphosphate receptors in signal transduction through the B‐cell antigen receptor , 1997, The EMBO journal.

[22]  Diana Murray,et al.  Membrane Targeting of C2 Domains of Phospholipase C-δ Isoforms* , 2002, The Journal of Biological Chemistry.

[23]  M. Iino,et al.  Transient Receptor Potential 1 Regulates Capacitative Ca2+ Entry and Ca2+ Release from Endoplasmic Reticulum in B Lymphocytes 〉 , 2002, The Journal of experimental medicine.

[24]  J. Putney.  PLC-γ: an old player has a new role , 2002, Nature Cell Biology.

[25]  RyujiInoue,et al.  The Transient Receptor Potential Protein Homologue TRP6 Is the Essential Component of Vascular α1-Adrenoceptor–Activated Ca2+-Permeable Cation Channel , 2001 .

[26]  J. Putney,et al.  The signal for capacitative calcium entry , 1993, Cell.

[27]  S. Rhee,et al.  Regulation of phosphoinositide-specific phospholipase C. , 2001, Annual review of biochemistry.

[28]  O. Petersen,et al.  Local and global cytosolic Ca2+ oscillations in exocrine cells evoked by agonists and inositol trisphosphate , 1993, Cell.

[29]  Keli Xu,et al.  Calcium oscillations increase the efficiency and specificity of gene expression , 1998, Nature.

[30]  S. Muallem,et al.  The N-terminal domain of the IP3 receptor gates store-operated hTrp3 channels. , 1999, Molecular cell.

[31]  Kyong-Tai Kim,et al.  Phospholipase C-δ1 Is Activated by Capacitative Calcium Entry That Follows Phospholipase C-β Activation upon Bradykinin Stimulation* , 1999, The Journal of Biological Chemistry.

[32]  K. Mikoshiba,et al.  Modulation of Ca(2+) entry by polypeptides of the inositol 1,4, 5-trisphosphate receptor (IP3R) that bind transient receptor potential (TRP): evidence for roles of TRP and IP3R in store depletion-activated Ca(2+) entry. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Kenzo Hirose,et al.  Visualization of IP3 Dynamics Reveals a Novel AMPA Receptor-Triggered IP3 Production Pathway Mediated by Voltage-Dependent Ca2+ Influx in Purkinje Cells , 2001, Neuron.

[34]  Anjana Rao,et al.  Gene regulation mediated by calcium signals in T lymphocytes , 2001, Nature Immunology.

[35]  J. Schlessinger,et al.  Activation of phospholipase Cγ by PI 3‐kinase‐induced PH domain‐mediated membrane targeting , 1998 .

[36]  A Bairoch,et al.  EF‐hand motifs in inositol phospholipid‐specific phospholipase C , 1990, FEBS letters.

[37]  S. Snyder,et al.  Phospholipase C-γ Is Required for Agonist-Induced Ca2+ Entry , 2002, Cell.

[38]  Y. Mori,et al.  Molecular and Functional Characterization of a Novel Mouse Transient Receptor Potential Protein Homologue TRP7 , 1999, The Journal of Biological Chemistry.

[39]  K. Venkatachalam,et al.  Expression of Functional Receptor-coupled TRPC3 Channels in DT40 Triple Receptor InsP3 knockout Cells* , 2001, The Journal of Biological Chemistry.

[40]  Y. Hara,et al.  The Transient Receptor Potential Protein Homologue TRP6 Is the Essential Component of Vascular &agr;1-Adrenoceptor–Activated Ca2+-Permeable Cation Channel , 2001, Circulation research.

[41]  N. Shimizu,et al.  LTRPC2 Ca2+-permeable channel activated by changes in redox status confers susceptibility to cell death. , 2002, Molecular cell.

[42]  Y. Fukami,et al.  Tyrosine kinase-dependent activation of phospholipase Cgamma is required for calcium transient in Xenopus egg fertilization. , 2000, Developmental biology.

[43]  J. Putney,et al.  An inositol 1,4,5‐trisphosphate receptor‐dependent cation entry pathway in DT40 B lymphocytes , 2002, The EMBO journal.

[44]  Kenzo Hirose,et al.  Encoding of Ca2+ signals by differential expression of IP3 receptor subtypes , 1999, The EMBO journal.

[45]  M. Negri,et al.  Activation of phospholipase Cγ by PI 3-kinase- induced PH domain-mediated membrane targeting , 1998 .

[46]  K. Mikoshiba,et al.  Modulation of Ca(2+) entry by polypeptides of the inositol 1,4, 5-trisphosphate receptor (IP3R) that bind transient receptor potential (TRP): evidence for roles of TRP and IP3R in store depletion-activated Ca(2+) entry. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[47]  C. Montell,et al.  The TRP Channels, a Remarkably Functional Family , 2002, Cell.

[48]  Roger Y. Tsien,et al.  Cell-permeant caged InsP3 ester shows that Ca2+ spike frequency can optimize gene expression , 1998, Nature.

[49]  T. Gudermann,et al.  Direct activation of human TRPC6 and TRPC3 channels by diacylglycerol , 1999, Nature.

[50]  Frederick Y. Wu,et al.  Phospholipase Cγ1 is a physiological guanine nucleotide exchange factor for the nuclear GTPase PIKE , 2002, Nature.

[51]  Andrew C. Chan,et al.  BLNK Required for Coupling Syk to PLCγ2 and Rac1-JNK in B Cells , 1999 .

[52]  R Y Tsien,et al.  Generation of calcium oscillations in fibroblasts by positive feedback between calcium and IP3. , 1991, Science.

[53]  S. Sprang,et al.  Structure of the first C2 domain of synaptotagmin I: A novel Ca2+/phospholipid-binding fold , 1995, Cell.

[54]  T. Kurosaki.  Regulation of B cell fates by BCR signaling components. , 2002, Current opinion in immunology.

[55]  M. Kano,et al.  Extracellular Calcium Controls the Dynamic Range of Neuronal Metabotropic Glutamate Receptor Responses , 2002, Molecular and Cellular Neuroscience.

[56]  Y. Mori,et al.  A new fluorescent biosensor for inositol trisphosphate. , 2002, Journal of the American Chemical Society.