Membrane-bound CTP:phosphocholine cytidylyltransferase regulates the rate of phosphatidylcholine synthesis in HeLa cells treated with unsaturated fatty acids.
The influence of fatty acids on phosphatidylcholine biosynthesis in HeLa cell cultures was investigated. Oleate and other unsaturated fatty acids stimulated the incorporation of phospho[Me-3H]choline into phosphatidylcholine from 5 to 20-fold, while saturated fatty acids were without effect. Stimulation of the reaction catalyzed by CTP:phosphocholine cytidylyltransferase (CTP:cholinephosphate cytidylyltransferase, EC 2.7.7.15) by 1 mM oleate was evident within 15 min and could be reversed within 40 min after removal of the oleate-supplemented cell medium. Cytidylyltransferase activity was 11-fold higher in homogenates from cells exposed to oleate. Treatment of the HeLa cells with oleate produced almost complete translocation of the cytidylyltransferase from the cytosol to the microsomal fraction. Additional support for conversion of the cytidylyltransferase to a membrane-bound form in oleate-treated cells was obtained in studies with digitonin. Exposure of control cells to digitonin for 2 min caused the release of 60% of the total cytidylyltransferase into the medium, while oleate-treated cells leaked only 5% of the enzyme in the presence of digitonin. Finally, oleate (50 microM) was shown to promote complete aggregation of cytosolic cytidylyltransferase with microsomes from HeLa cells and 22-fold stimulation of the enzyme activity. It appears that the rate of phosphatidylcholine biosynthesis is governed by the amount of cytidylyltransferase bound to endoplasmic reticulum in HeLa cells.
D002794:Choline / Q000378:metabolism; D019916:Choline-Phosphate Cytidylyltransferase; D004072:Digitonin / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D009713:Nucleotidyltransferases / Q000378:metabolism*; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000201:enzymology; D014316:Tritium
D005231:Fatty Acids, Unsaturated; D009829:Oleic Acids; D010713:Phosphatidylcholines; D014316:Tritium; D019301:Oleic Acid; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase; D004072:Digitonin; D002794:Choline