Rapid activation of phosphatidate phosphohydrolase in mesangial cells by lipid A.
Knowledge of rapid events in cell signaling initiated by lipid A, the core moiety of bacterial lipopolysaccharide, is limited. In the present study we have demonstrated that cis-parinaric acid (cis-PnA) rapidly labels 1,2-sn-diacylglycerol (DAG) subsequent to labeling of phosphatidic acid (PA). Stimulation of microsomal membranes with lipid A decreased the level of PA labeled with cis-PnA within 5 s and increased the proportion of fluorescent label in DAG. Lipid A stimulation of DAG synthesis at 5-15 s was inhibited by incubation of mesangial cells with pertussis toxin prior to isolation of microsomal membranes. Inhibition of DAG formation was accompanied by an accumulation of the mass and fluorescent label in the cis-PnA-labeled phosphatidic acid pool. GTP gamma S caused a decrease in labeled PA and an increase in labeled 1,2-DAG. We conclude that the PA pool was enlarged via the lipid A sensitive lyso-PA acyl transferase (lyso-PA-AT) and was decreased by a phosphatidate phosphohydrolase to form DAG. The phosphatidate phosphohydrolase was at least partly regulated by a pertussis-sensitive G-protein. Lipid A or 1,2-dilinoleyl-PA, a product of lyso-PA-AT, induced cell activation as monitored by actin reorganization and cellular shape changes. Pretreatment of cells with pertussis toxin prevented the morphological changes normally induced by lipid A or 1,2-dilinoleyl-PA. In contrast, 1-oleoyl-2-acetylglycerol induced rapid actin reorganization and shape change, presumably bypassing the pertussis blockade. We propose that specific pools of PA and PA-derived DAG are key elements in rapid signaling in mesangial cells and are independent of the PI cycle and phospholipase C.
Bursten
S L
SL
Department of Medicine, University of Washington, Seattle 98195.
Harris
W E
WE
D000217:Acyltransferases / Q000494:pharmacology; D000818:Animals; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000494:pharmacology; D005920:Glomerular Mesangium / Q000187:drug effects / Q000201:enzymology*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D008050:Lipid A / Q000494:pharmacology*; D008861:Microsomes / Q000201:enzymology; D037342:Pertussis Toxin; D010711:Phosphatidate Phosphatase / Q000378:metabolism*; D010712:Phosphatidic Acids / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D004075:Diglycerides; D008050:Lipid A; D010712:Phosphatidic Acids; D010743:Phospholipids; D010566:Virulence Factors, Bordetella; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D000217:Acyltransferases; C018764:2-acylglycerophosphate acyltransferase; D037342:Pertussis Toxin; D010711:Phosphatidate Phosphatase; D019204:GTP-Binding Proteins