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- Title
Phospholipase C, Protein Kinase C, Ca<sup>2+</sup>/Calmodulin-Dependent Protein Kinase II, and Tyrosine Phosphorylation Are Involved in Carbachol-Induced Phospholipase D Activation in Chinese Hamster Ovary Cells Expressing Muscarinic Acetylcholine Receptor of Caenorhabditis elegans.
- Authors
Min, Do Sik; Cho, Nam Jeong; Yoon, Shin Hee; Lee, Young Han; Hahn, Sang-June; Lee, Kweon-Haeng; Kim, Myung-Suk; Jo, Yang-Hyeok
- Abstract
Abstract: Recently, we have isolated a cDNA encoding a muscarinic acetylcholine receptor (mAChR) from Caenorhabditis elegans. To investigate the regulation of phospholipase D (PLD) signaling via a muscarinic receptor, we generated stable transfected Chinese hamster ovary (CHO) cells that overexpress the mAChR of C. elegans (CHO-GAR-3). Carbachol (CCh) induced inositol phosphate formation and a significantly higher Ca2+ elevation and stimulated PLD activity through the mAChR ; this was insensitive to pertussis toxin, but its activity was abolished by the phospholipase C (PLC) inhibitor U73122. Western blot analysis revealed several apparent tyrosine-phosphorylated protein bands after CCh treatment. The CCh-induced PLD activation and tyrosine phosphorylation were significantly reduced by the protein kinase C (PKC) inhibitor calphostin C and down-regulation of PKC and the tyrosine kinase inhibitor genistein. Moreover, the Ca2+-calmodulin-dependent protein kinase II (CaM kinase II) inhibitor KN62, in addition to chelation of extracellular or intracellular Ca2+ by EGTA and BAPTA/AM, abolished CCh-induced PLD activation and protein tyrosine phosphorylation. Taken together, these results suggest that the PLC/PKC-PLD pathway and the CaM kinase II/tyrosine kinase-PLD pathway are involved in the activation of PLD through mAChRs of C. elegans.
- Subjects
ANTISENSE DNA; MUSCARINIC receptors; PHOSPHOLIPASE C; PROTEIN kinase C; CAENORHABDITIS elegans
- Publication
Journal of Neurochemistry, 2000, Vol 75, Issue 1, p274
- ISSN
0022-3042
- Publication type
Article
- DOI
10.1046/j.1471-4159.2000.0750274.x