title: ADP-ribosylation factor-dependent phospholipase D activation by the M3 muscarinic receptor.
abstract: G protein-coupled receptors can potentially activate phospholipase D (PLD) by a number of routes. We show here that the native M3 muscarinic receptor in 1321N1 cells and an epitope-tagged M3 receptor expressed in COS7 cells substantially utilize an ADP-ribosylation factor (ARF)-dependent route of PLD activation. This pathway is activated at the plasma membrane but appears to be largely independent of G, phospholipase C, Ca2+ q/11, protein kinase C, tyrosine kinases, and phosphatidyl inositol 3-kinase. We report instead that it involves physical association of ARF with the M3 receptor as demonstrated by co-immunoprecipitation and by in vitro interaction with a glutathione S-transferase fusion protein of the receptor's third intracellular loop domain. Experiments with mutant constructs of ARF1/6 and PLD1/2 indicate that the M3 receptor displays a major ARF1-dependent route of PLD1 activation with an additional ARF6-dependent pathway to PLD1 or PLD2. Examples of other G protein-coupled receptors assessed in comparison display alternative pathways of protein kinase C- or ARF6-dependent activation of PLD2.
authors.affiliation: Medical Research Council Membrane and Adapter Proteins Co-operative Group, Membrane Biology Interdisciplinary Research Group, School of Biomedical and Clinical Laboratory Sciences, University of Edinburgh, EH8 9XD, United Kingdom. Rory.Mitchell@ed.ac.uk
authors.forename: Rory
authors.initials: R
authors.lastname: Mitchell
authors.forename: Derek N
authors.initials: DN
authors.lastname: Robertson
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Holland
authors.forename: Daniel
authors.initials: D
authors.lastname: Collins
authors.forename: Eve M
authors.initials: EM
authors.lastname: Lutz
authors.forename: Melanie S
authors.initials: MS
authors.lastname: Johnson
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000378:metabolism; D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D000818:Animals; D019921:Biotinylation; D015153:Blotting, Western; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D002217:Carbachol / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000939:Epitopes; D004963:Estrenes / Q000494:pharmacology; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D020128:Inhibitory Concentration 50; D008024:Ligands; D008954:Models, Biological; D009154:Mutation; D010739:Phospholipase D / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011760:Pyrrolidinones / Q000494:pharmacology; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic / Q000378:metabolism*; D015398:Signal Transduction; D013347:Subcellular Fractions; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D000089982:ADP-Ribosylation Factor 6; D004791:Enzyme Inhibitors; D000939:Epitopes; D004963:Estrenes; D008024:Ligands; D011760:Pyrrolidinones; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D020126:Brefeldin A; D002217:Carbachol; D005982:Glutathione Transferase; D011493:Protein Kinase C; D010739:Phospholipase D; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human