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Involvement of phosphatidylinositol 3'-kinase in stem-cell-factor-induced phospholipase D activation and arachidonic acid release

Kozawa, Osamu ; Blume-Jensen, Peter ; Heldin, Carl-Henrik and Rönnstrand, Lars LU (1997) In European Journal of Biochemistry 248(1). p.149-155
Abstract
We have shown previously that the stem cell factor (SCF) receptor undergoes phosphorylation on serine residues following ligand stimulation, and that this phopshorylation is dependent mainly on the activity of protein kinase C (PKC). In the present study, we have further investigated the molecular mechanisms behind SCF-stimulated activation of PKC, and found that SCF does not activate phosphatidylinositol-specific phospholipase C. In contrast, phospholipase D (PLD) is activated in response to SCF in a dose-dependent manner. Activation of PLD was not inhibited by calphostin C, an inhibitor of PKC. On the other hand, inhibitors of phosphatidylinositol PtdIns 3'-kinase (PtdIns 3'-kinase), i.e. wortmannin and LY294002, inhibited SCF-induced... (More)
We have shown previously that the stem cell factor (SCF) receptor undergoes phosphorylation on serine residues following ligand stimulation, and that this phopshorylation is dependent mainly on the activity of protein kinase C (PKC). In the present study, we have further investigated the molecular mechanisms behind SCF-stimulated activation of PKC, and found that SCF does not activate phosphatidylinositol-specific phospholipase C. In contrast, phospholipase D (PLD) is activated in response to SCF in a dose-dependent manner. Activation of PLD was not inhibited by calphostin C, an inhibitor of PKC. On the other hand, inhibitors of phosphatidylinositol PtdIns 3'-kinase (PtdIns 3'-kinase), i.e. wortmannin and LY294002, inhibited SCF-induced PLD activation. Moreover, a mutant SCF receptor in which Tyr721, which is responsible for activation of PtdIns 3'-kinase, is mutated to a phenylalanine residue was unable to mediate activation of PLD. Thus, PtdIns 3'-kinase appears to be essential for SCF-induced PLD activation. Furthermore, we demonstrate that phosphatidic acid (PtdH), generated through the action of PLD in response to SCF, is metabolized to diacylglycerol by dephosphorylation. Diacylglycerol can then activate PKC, and, moreover, after deacylation by a diacylglycerol lipase, yield arachidonic acid, an important second messenger in cell signaling. (Less)
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subject
keywords
Complementary/genetics Diglycerides/biosynthesis Enzyme Activation/drug effects Humans Models, Arachidonic Acid/*metabolism Base Sequence Cell Line DNA, Biological Mutagenesis, Site-Directed Phosphatidylinositol 3-Kinases Phosphatidylinositol Diacylglycerol-Lyase Phosphoinositide Phospholipase C Phospholipase D/*metabolism Phosphotransferases (Alcohol Group Acceptor)/*metabolism Propranolol/pharmacology Proto-Oncogene Proteins c-kit/genetics/metabolism Second Messenger Systems Stem Cell Factor/metabolism/*pharmacology Transfection Type C Phospholipases/metabolism
in
European Journal of Biochemistry
volume
248
issue
1
pages
149 - 155
publisher
Wiley-Blackwell
external identifiers
  • scopus:0030835325
ISSN
0014-2956
DOI
10.1111/j.1432-1033
language
English
LU publication?
no
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The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Experimental Clinical Chemistry (013016010)
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06fc0d60-e8bb-4b48-afea-27c32c777809 (old id 1783916)
alternative location
http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1997.00149.x/full
date added to LUP
2016-04-04 09:30:23
date last changed
2020-01-12 20:56:23
@article{06fc0d60-e8bb-4b48-afea-27c32c777809,
  abstract     = {We have shown previously that the stem cell factor (SCF) receptor undergoes phosphorylation on serine residues following ligand stimulation, and that this phopshorylation is dependent mainly on the activity of protein kinase C (PKC). In the present study, we have further investigated the molecular mechanisms behind SCF-stimulated activation of PKC, and found that SCF does not activate phosphatidylinositol-specific phospholipase C. In contrast, phospholipase D (PLD) is activated in response to SCF in a dose-dependent manner. Activation of PLD was not inhibited by calphostin C, an inhibitor of PKC. On the other hand, inhibitors of phosphatidylinositol PtdIns 3'-kinase (PtdIns 3'-kinase), i.e. wortmannin and LY294002, inhibited SCF-induced PLD activation. Moreover, a mutant SCF receptor in which Tyr721, which is responsible for activation of PtdIns 3'-kinase, is mutated to a phenylalanine residue was unable to mediate activation of PLD. Thus, PtdIns 3'-kinase appears to be essential for SCF-induced PLD activation. Furthermore, we demonstrate that phosphatidic acid (PtdH), generated through the action of PLD in response to SCF, is metabolized to diacylglycerol by dephosphorylation. Diacylglycerol can then activate PKC, and, moreover, after deacylation by a diacylglycerol lipase, yield arachidonic acid, an important second messenger in cell signaling.},
  author       = {Kozawa, Osamu and Blume-Jensen, Peter and Heldin, Carl-Henrik and Rönnstrand, Lars},
  issn         = {0014-2956},
  language     = {eng},
  number       = {1},
  pages        = {149--155},
  publisher    = {Wiley-Blackwell},
  series       = {European Journal of Biochemistry},
  title        = {Involvement of phosphatidylinositol 3'-kinase in stem-cell-factor-induced phospholipase D activation and arachidonic acid release},
  url          = {http://dx.doi.org/10.1111/j.1432-1033},
  doi          = {10.1111/j.1432-1033},
  volume       = {248},
  year         = {1997},
}