Sustained Activation of the Tyrosine Kinase Syk by Antigen in Mast Cells Requires Local Ca2+ Influx through Ca2+ Release-activated Ca2+ Channels

Ng, Siaw Wei; Capite, Joseph Di; Singaravelu, Karthika; Parekh, Anant B.

doi:10.1074/jbc.m804942200

Cited by 105 publications

(97 citation statements)

References 47 publications

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“…Transfection-RNAi to Syk (Invitrogen) and scrambled RNAi were transfected using the AMAXA system (21). The Syk construct was as follows (5Ј-3Ј): sense, CCCUCUGG-CAGCUAGUGGAACAUUA; antisense, UAAUGUUCCACUAGC-UGCCAGAGGG.…”

Section: Methodsmentioning

confidence: 99%

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Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Nelson

Parekh

2009

Journal of Biological Chemistry

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microdomain, which occurs following the opening of a Ca 2ϩ -permeable channel in either the plasma membrane or intracellular organelles (5). Because the volume a microdomain occupies is extremely small, the Ca 2ϩ concentration can rise to reach levels that are orders of magnitude greater than the bulk cytoplasmic Ca 2ϩ rise (3, 5). Store-operated Ca 2ϩ channels are the major route for agonist-evoked Ca 2ϩ entry in non-excitable cells and open following stimulation of phospholipase C-coupled receptors (6). These receptors generate inositol 1,4,5-trisphosphate, which releases Ca 2ϩ from the endoplasmic reticulum (ER) 3 (7). The fall in Ca 2ϩ content within the store is detected by the ER Ca 2ϩ sensor STIM1, which migrates to specialized ER-plasma membrane junctions, where it opens the storeoperated channels (8). The best characterized store-operated channel is the Ca 2ϩ release-activated Ca 2ϩ (CRAC) channel (6), the pore-forming subunit of which is composed of Orai1 (9 -12). Ca 2ϩ entry through CRAC channels regulates enzyme activity, secretion, gene expression, and cell growth and proliferation (13).Here, we have examined whether excitation-transcription coupling is driven by Ca 2ϩ microdomains arising from open CRAC channels in mast cells. In T cells, NFAT-dependent gene expression requires a global Ca 2ϩ rise (14 -16). We find that local Ca 2ϩ influx signals to the nucleus much more effectively than a robust bulk Ca 2ϩ rise. This leads to the expression of the transcription factor c-fos, a regulator of proinflammatory gene expression (17). Furthermore, the non-receptor tyrosine kinase Syk clusters at the cell periphery and couples Ca 2ϩ microdomains to c-fos expression through recruitment of the cytoplasmic transcription factor STAT5 in a protein kinase C-and MEK/ERK-independent pathway. Ca 2ϩ microdomains following CRAC channel activation also activate Ca 2ϩ -dependent phospholipase A 2 , followed by secretion of cysteinyl leukotrienes (18). However, unlike c-fos expression, this is mediated via the MEK/ERK pathway. Parallel processing of the Ca 2ϩ microdomain by Syk through two distinct signaling pathways constitutes a novel mechanism to evoke spatially and temporally different cellular responses. EXPERIMENTAL PROCEDURESCell Culture-Rat basophilic leukemia-1 (RBL-1), an immortalized mast cell line, was bought from ATCC. Cells were cultured (37°C, 5% CO 2 ) in Dulbecco's modified Eagle's medium with 10% fetal bovine serum, 2 mM L-glutamine, and penicillin/ streptomycin, as described previously (19). For Ca 2ϩ imaging and patch clamp experiments, cells were passaged (using trypsin) onto glass coverslips and used 24 -48 h after plating. All cells were used between passages 4 and 16.Ca 2ϩ Imaging-Ca 2ϩ imaging experiments were carried out using the IMAGO CCD camera-based system from TILL Photonics, as described previously using Fura 5F (loaded in the AM form) (18). Cells were alternately excited at 356 and 380 nm (20-ms exposures; 0.5 Hz) using a polychrome monochromator. Images were analyzed offline using IGOR ...

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Section: Methodsmentioning

confidence: 99%

“…In some experiments, the cytoplasm was loaded with EGTA or BAPTA through the AM derivative. Loading was exactly as described previously (21).…”

Section: Methodsmentioning

confidence: 99%

Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Nelson

Parekh

2009

Journal of Biological Chemistry

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“…By contrast, activation of FCεRI receptors with antigen or IgE leads to a Ca 2+ response that develops after a longer delay and which is punctuated by a series of slow Ca 2+ oscillations on an elevated background Ca 2+ rise (15,16). Both types of Ca 2+ response depend on Ca 2+ influx through CRAC channels: Both agonists activate CRAC channels in RBL cells (16,17), and exposure to CRAC channel blockers accelerates decline of the Ca 2+ signal in response to either cysLT1 (7) or FCεRI receptor activation (18). However, the kinetics of InsP 3 production and steady-state InsP 3 levels differ between the stimuli.…”

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Different agonists recruit different stromal interaction molecule proteins to support cytoplasmic Ca ²⁺ oscillations and gene expression

Kar

Bakowski

Capite

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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100

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Stimulation of cells with physiological concentrations of calciummobilizing agonists often results in the generation of repetitive cytoplasmic Ca 2+ oscillations. Although oscillations arise from regenerative Ca 2+ release, they are sustained by store-operated Ca 2+ entry through Ca 2+ release-activated Ca 2+ (CRAC) channels. Here, we show that following stimulation of cysteinyl leukotriene type I receptors in rat basophilic leukemia (RBL)-1 cells, large amplitude Ca 2+ oscillations, CRAC channel activity, and downstream Ca 2+ -dependent nuclear factor of activated T cells (NFAT)-driven gene expression are all exclusively maintained by the endoplasmic reticulum Ca 2+ sensor stromal interaction molecule (STIM) 1. However, stimulation of tyrosine kinase-coupled FCεRI receptors evoked Ca 2+ oscillations and NFAT-dependent gene expression through recruitment of both STIM2 and STIM1. We conclude that different agonists activate different STIM proteins to sustain Ca 2+ signals and downstream responses.excitation-transcription coupling | transcription S timulation of cell-surface receptors that couple to the phospholipase C pathway with physiological concentrations of agonist generally evokes repetitive cytoplasmic Ca 2+ oscillations (1). Oscillatory Ca 2+ signals enable cytoplasmic Ca 2+ to reach high levels transiently, thereby avoiding the deleterious effects of a prolonged, elevated Ca 2+ rise. Information is encoded in the oscillatory amplitude and frequency (2) and the spatial profile of the Ca 2+ signal (3), each of which can be deciphered by cells to drive selective downstream responses.Ca 2+ oscillations are triggered by inositol trisphosphate (InsP 3 )-mediated Ca 2+ release from intracellular Ca 2+ stores, primarily the endoplasmic reticulum (ER) (2). The resulting fall in Ca 2+ within the stores opens store-operated CRAC channels in the plasma membrane (4, 5). Ca 2+ entry through these channels refills the stores and, thus, sustains InsP 3 -dependent Ca 2+ oscillations (6). In addition to this supportive role, local Ca 2+ entry through Ca 2+ release-activated Ca 2+ (CRAC) channels during oscillatory responses in mast cells, and not the oscillations per se, signals to the nucleus to regulate Ca 2+ -dependent gene expression (7).The two main molecular components of store-operated Ca 2+ entry are the stromal interaction molecule (STIM) and Orai proteins (reviewed in refs. 8-10). The transmembrane ER proteins STIM1 and STIM2 detect ER Ca 2+ content through an EF-hand domain in their respective N-termini, which face the lumen of the store. Loss of luminal Ca 2+ leads to STIM aggregation within the ER followed by migration to ER-plasma membrane junctions located just below the plasma membrane. Here, they bind to and activate Orai1, a four transmembrane domain spanning plasma membrane protein, which forms the CRAC channel.Despite significant homology between STIM1 and STIM2, there are some important differences between them. First, they differ in their respective abilities to activate Orai1. STIM2 activates Ca 2+ ...

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“…In view of the complexities of pancreatic acinar cell Ca 2+ signaling in physiology and pathology and, in particular, the uncertainty about the nature of the Ca 2+ entry pathways, it is unknown whether blockade of Ca 2+ -selective CRAC channels could in principle be effective as therapy against acute pancreatitis. With the availability of relatively specific CRAC channel blockers (30)(31)(32)(33)(34) there is now an opportunity to test the hypothesis that one type of Ca 2+ entry channel is so important that pharmacological blockade could afford effective protection against toxic Ca 2+ signal generation and its consequences in intact pancreatic acinar cells. Given the current absence of any specific therapy against pancreatitis, a demonstration that pharmacological blockade of CRAC channels could prevent the dangerous necrosis evoked by agents known to initiate pancreatitis, would be a proof of principle that a specific therapy could be developed.…”

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confidence: 99%

Ca ²⁺ release-activated Ca ²⁺ channel blockade as a potential tool in antipancreatitis therapy

Gerasimenko

Gryshchenko

Ferdek

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

171

225

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Alcohol-related acute pancreatitis can be mediated by a combination of alcohol and fatty acids (fatty acid ethyl esters) and is initiated by a sustained elevation of the Ca 2+ concentration inside pancreatic acinar cells ([Ca 2+ ] i ), due to excessive release of Ca 2+ stored inside the cells followed by Ca 2+ entry from the interstitial fluid. The sustained [Ca 2+ ] i elevation activates intracellular digestive proenzymes resulting in necrosis and inflammation. We tested the hypothesis that pharmacological blockade of store-operated or Ca 2+ release-activated Ca 2+ channels (CRAC) would prevent sustained elevation of [Ca 2+ ] i and therefore protease activation and necrosis. In isolated mouse pancreatic acinar cells, CRAC channels were activated by blocking Ca 2+ ATPase pumps in the endoplasmic reticulum with thapsigargin in the absence of external Ca 2+ . Ca 2+ entry then occurred upon admission of Ca 2+ to the extracellular solution. The CRAC channel blocker developed by GlaxoSmithKline, GSK-7975A, inhibited store-operated Ca 2+ entry in a concentrationdependent manner within the range of 1 to 50 μM (IC 50 = 3.4 μM), but had little or no effect on the physiological Ca 2+ spiking evoked by acetylcholine or cholecystokinin. Palmitoleic acid ethyl ester (100 μM), an important mediator of alcohol-related pancreatitis, evoked a sustained elevation of [Ca 2+ ] i , which was markedly reduced by CRAC blockade. Importantly, the palmitoleic acid ethyl ester-induced trypsin and protease activity as well as necrosis were almost abolished by blocking CRAC channels. There is currently no specific treatment of pancreatitis, but our data show that pharmacological CRAC blockade is highly effective against toxic [Ca 2+ ] i elevation, necrosis, and trypsin/protease activity and therefore has potential to effectively treat pancreatitis.capacitative Ca 2+ entry | alcohol metabolite | pancreas | hepatocyte Ca 2+ entry | AR42JA cute pancreatitis is a human disease mostly caused by alcohol abuse or complications from biliary disease. In this disease, against which there is currently no effective therapy, digestive proenzymes are prematurely activated inside the acinar cells leading to autodigestion and necrosis (1-3). Intracellular Ca 2+ plays a critical role in the initiation of this disease process (2-4), but intracellular Ca 2+ also plays a critical role in the physiological regulation of the normal exocytotic secretion of the digestive proenzymes (5).The pancreatic acinar cells are capable of generating multiple patterns of cytosolic Ca 2+ signals depending on the type and concentration of the stimulating agent (5). The physiological Ca 2+ signals regulating secretion-evoked by the neurotransmitter acetylcholine (ACh) or the hormone cholecystokinin (CCK)-consist of repetitive short-lasting rises in the cytosolic Ca 2+ concentration ([Ca 2+ ] i ). These are mostly confined to the apical area, in which the secretory (zymogen) granules (ZGs) are concentrated, by a belt of perigranular mitochondria operating as a firewall against...

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Sustained Activation of the Tyrosine Kinase Syk by Antigen in Mast Cells Requires Local Ca2+ Influx through Ca2+ Release-activated Ca2+ Channels

Cited by 105 publications

References 47 publications

Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Different agonists recruit different stromal interaction molecule proteins to support cytoplasmic Ca ²⁺ oscillations and gene expression

Ca ²⁺ release-activated Ca ²⁺ channel blockade as a potential tool in antipancreatitis therapy

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Sustained Activation of the Tyrosine Kinase Syk by Antigen in Mast Cells Requires Local Ca2+ Influx through Ca2+ Release-activated Ca2+ Channels

Cited by 105 publications

References 47 publications

Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Coupling of Ca2+ Microdomains to Spatially and Temporally Distinct Cellular Responses by the Tyrosine Kinase Syk

Different agonists recruit different stromal interaction molecule proteins to support cytoplasmic Ca 2+ oscillations and gene expression

Ca 2+ release-activated Ca 2+ channel blockade as a potential tool in antipancreatitis therapy

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Different agonists recruit different stromal interaction molecule proteins to support cytoplasmic Ca ²⁺ oscillations and gene expression

Ca ²⁺ release-activated Ca ²⁺ channel blockade as a potential tool in antipancreatitis therapy