title: Negative feedback regulation of Gq signaling by protein kinase C is disrupted by diacylglycerol kinase ζ in COS-7 cells.
abstract: Cellular response to G(q)-linked agonists is shaped by regulatory inputs which determine signal strength and duration. Stimulation of phospholipase C-β (PLC-β) lipase activity results in an increase in the levels of diacylglycerol (DAG) and activation of protein kinase C (PKC) activity. PKC has been implicated in the feedback regulation of G(q) signaling through actions on PLC-β and phospholipase D (PLD) lipase activity. As PKC activity is modulated by multiple layers of regulation, the physiological impact of PKC on G(q) signaling is unclear. PKC signaling can be terminated by diacylglycerol kinases (DGKs) which are regulated in a cell-specific manner. The present studies investigated the contribution of the ubiquitously expressed DGKζ isoform in the regulation of PKC signaling and G(q) response in transfected COS-7 cells. Genetic depletion of DGKζ protein with antisense oligonucleotides dramatically reduced DAG metabolism. The sustained increase in PKC signaling was associated with a pronounced inhibition of carbachol-stimulated lipase activity in cells co-transfected with m1 muscarinic receptor, Gα(q) and either with or without PLC-β(1). The data also reveal that sustained activation of PKC alone does not increase cellular PLD1 activity. Therefore, G(12)-activated RhoA is physiologically important for adequate stimulation of PLD1 activity. These data show that the impact of PKC on G(q) signal transduction is determined by the background of cell-specific processes.
authors.affiliation: Department of Molecular and Cellular Pharmacology, University of Miami Miller School of Medicine, FL 33101-6189, USA. ilitosch@med.miami.edu
authors.forename: Irene
authors.initials: I
authors.lastname: Litosch
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D019852:Diacylglycerol Kinase / Q000235:genetics / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism; D025461:Feedback, Physiological; D043802:GTP-Binding Protein alpha Subunits, Gq-G11 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010739:Phospholipase D / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D064546:Protein Kinase C beta; D043643:Receptor, Muscarinic M1 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D004075:Diglycerides; D043643:Receptor, Muscarinic M1; C526463:DGKZ protein, human; D019852:Diacylglycerol Kinase; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D010739:Phospholipase D; C106801:phospholipase D1; D043802:GTP-Binding Protein alpha Subunits, Gq-G11