Regulation of phosphatidylcholine biosynthesis in cultured chick embryonic muscle treated with phospholipase C.
Cultures of embryonic chick muscle cells grown in medium containing phospholipase C from Clostridium perfringens incorporated [3H]choline into lipid at a rate 3- to 5-fold higher than control cultures. To determine the mechanism by which stimulation of phosphatidylcholine synthesis occurred in phospholipase C-treated cells, activities of enzymes and levels of intermediates in the biosynthetic pathway for phosphatidylcholine were examined. Activities of choline kinase, choline phosphotransferase, glycerol-3-phosphate dehydrogenase, glycerol-3-phosphate acyltransferase, acylglycerol-3-phosphate acyltransferase, and phosphatidic acid phosphatase in phospholipase C-treated cells were the same or only slightly higher than in control cells. CTP:phosphocholine cytidylyltransferase, on the other hand, was 3 times as active in homogenates from phospholipase C-treated cells. Levels of phosphocholine decreased and levels of CDP-choline increased in phospholipase C-treated cells, and a calculation of the disequilibrium ratio indicated that the cytidylyltransferase reaction was not at equilibrium. The cytidylyltransferase was, thus, identified as the regulatory enzyme for choline flux in these cells. The cytidylyltransferase was located in both the cytosolic and particulate fractions from cultured muscle cells and a much larger portion of enzyme activity was associated with the particulate fraction in cells treated with phospholipase C. Sonicated preparations of total chick lipids, phosphatidylethanolamine, and phosphatidylserine greatly stimulated the cytosolic cytidylyltransferase activity but had no effect on the particulate enzyme. Neither stimulation of incorporation of [3H]choline into lipid nor activation of the cytidylyltransferase was dependent on protein synthesis. A model for the mechanism of regulation of phosphatidylcholine synthesis in embryonic chick muscle is presented.
D000818:Animals; D002642:Chick Embryo; D002794:Choline / Q000378:metabolism; D019916:Choline-Phosphate Cytidylyltransferase; D003513:Cycloheximide / Q000494:pharmacology; D003566:Cytidine Diphosphate Choline / Q000378:metabolism; D007700:Kinetics; D009132:Muscles / Q000187:drug effects / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010714:Phosphatidylethanolamines / Q000096:biosynthesis; D010740:Phospholipases / Q000494:pharmacology*; D010738:Type C Phospholipases / Q000494:pharmacology*
D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D003566:Cytidine Diphosphate Choline; D003513:Cycloheximide; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase; D010740:Phospholipases; D010738:Type C Phospholipases; D002794:Choline