Contribution of lipid mediators to the regulation of phosphatidylcholine synthesis by angiotensin.
Angiotensin stimulates a cellular mitogenic response via the AT1 receptor. We have examined the effect of angiotensin on the rate of phosphatidylcholine (PC) synthesis and have begun to dissect the pathway linking the AT1 receptor to the rate-limiting enzyme in PC synthesis, CTP: phosphocholine cytidylyltransferase (CCT), using CHO cells engineered to express the AT1a receptor. Since CCT can be directly activated by lipid mediators, we probed for their involvement in the PC synthesis response to angiotensin. Angiotensin stimulated CCT activity and PC synthesis two- to threefold after a 30-min delay. The kinetics of this stimulation most closely paralleled an increase in diacylglycerol (DAG) derived from myristic acid-enriched phospholipids. The production of arachidonic acid, phosphatidic acid, or reactive oxygen species either peaked much earlier or not at all. Moreover, manipulation of the intracellular supply of oxygen free radicals, arachidonic acid, HETEs, or phosphatidic acid (using inhibitors and/or exogenous addition) did not generate parallel effects on the rate of PC synthesis. Restricting the production of DAG by inhibition of PLCbeta with U73122 reduced both basal and angiotensin-stimulated PC synthesis. The U73122 inhibition of PC synthesis was accompanied by a similar inhibition of ERK1/2 phosphorylation. Addition of exogenous DAG stimulated basal and angiotensin-dependent PC synthesis, and partially reversed the effect of the PLC inhibitor on PC synthesis. These results do not provide support for lipid mediators as direct stimulators of CCT and PC synthesis downstream of angiotensin, but give rise to the idea that angiotensin effects might be mediated via ERK1/2.
Department of Molecular Biology and Biochemistry, Simon Fraser University, Burnaby, BC, Canada V5A 1S6.
Theresa E
TE
Kitos
Adrienne
A
Drobnies
Michael N P
MN
Ng
Yeshao
Y
Wen
Rosemary B
RB
Cornell
D000809:Angiotensins / Q000494:pharmacology*; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D016466:CHO Cells; D019916:Choline-Phosphate Cytidylyltransferase / Q000378:metabolism; D006224:Cricetinae; D004075:Diglycerides / Q000378:metabolism; D004963:Estrenes / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism; D007700:Kinetics; D050356:Lipid Metabolism / Q000187:drug effects; D010712:Phosphatidic Acids / Q000378:metabolism; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D011760:Pyrrolidinones / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000809:Angiotensins; D004075:Diglycerides; D004963:Estrenes; D006893:Hydroxyeicosatetraenoic Acids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D011760:Pyrrolidinones; D017382:Reactive Oxygen Species; D044140:Receptor, Angiotensin, Type 1; D011994:Recombinant Proteins; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D016718:Arachidonic Acid; D048049:Extracellular Signal-Regulated MAP Kinases; D019916:Choline-Phosphate Cytidylyltransferase