Phospholipase C, protein kinase C, Ca2+/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

Citation
Ds. Min et al., Phospholipase C, protein kinase C, Ca2+/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, J NEUROCHEM, 75(1), 2000, pp. 274-281
Citations number
35
Categorie Soggetti
Neurosciences & Behavoir
Journal title
JOURNAL OF NEUROCHEMISTRY
ISSN journal
00223042 → ACNP
Volume
75
Issue
1
Year of publication
2000
Pages
274 - 281
Database
ISI
SICI code
0022-3042(200007)75:1<274:PCPKCC>2.0.ZU;2-M
Abstract
Recently, we have isolated a cDNA encoding a muscarinic acetylcholine recep tor (mAChR) from Caenorhabditis elegans. To investigate the regulation of p hospholipase D (PLD) signaling via a muscarinic receptor, we generated stab le transfected Chinese hamster ovary (CHO) cells that overexpress the mAChR of C. elegans (CHO-GAR-3). Carbachol (CCh) induced inositol phosphate form ation and a significantly higher Ca2+ elevation and stimulated PLD activity through the mAChR; this was insensitive to pertussis toxin, but its activi ty was abolished by the phospholipase C (PLC) inhibitor U73122. Western blo t analysis revealed several apparent tyrosine-phosphorylated protein bands after CCh treatment. The CCh-induced PLD activation and tyrosine phosphoryl ation were significantly reduced by the protein kinase C (PKC) inhibitor ca lphostin C and down-regulation of PKC and the tyrosine kinase inhibitor gen istein. Moreover, the Ca2+/calmodulin-dependent protein kinase If (CaM kina se II) inhibitor KN62, in addition to chelation of extracellular or intrace llular Ca2+ by EGTA and BAPTA/AM, abolished CCh-induced PLD activation and protein tyrosine phosphorylation. Taken together, these results suggest tha t the PLC/PKC-PLD pathway and the CaM kinase Il/tyrosine kinase-PLD pathway are involved in the activation of PLD through mAChRs of C. elegans.