Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1993 Apr 15;291(Pt 2):419–427. doi: 10.1042/bj2910419

Evidence that binding of CTP:phosphocholine cytidylyltransferase to membranes in rat hepatocytes is modulated by the ratio of bilayer- to non-bilayer-forming lipids.

H Jamil 1, G M Hatch 1, D E Vance 1
PMCID: PMC1132542  PMID: 8387269

Abstract

The mechanism by which phospholipase C (PLC) digestion of cultured cells mediates binding of CTP:phosphocholine cytidylyltransferase to cellular membranes was investigated. Incubation of choline-depleted rat hepatocytes with PLC caused a translocation of enzyme from cytosol to membranes concomitant with a decrease in the concentration of phosphatidylcholine with no effect on the concentration of other phospholipids. Removal of PLC and supplementation with choline restored the amount of phosphatidylcholine in the cells and translocated cytidylyltransferase to the cytosol. However, when phosphatidylcholine levels were decreased by incubation with phospholipase A2 (PLA2), there was no significant redistribution of cytidylyltransferase activity. With PLA2 the concentration of phosphatidylethanolamine, as well as of phosphatidylcholine, was significantly decreased. Since PLC, but not phospholipase A2, raised the cellular concentration of diacylglycerol, possibly diacylglycerol mediated the binding of cytidylyltransferase to membranes. This possibility was examined, but is unlikely, since addition of lysophosphatidylcholine to PLC-treated cells restored the concentration of phosphatidylcholine and released cytidylyltransferase into the cytosol, but did not lower diacylglycerol levels to normal values. Studies in vitro, incubations of cells with choline analogues and a survey of the literature suggested that the over-riding common factor in regulation of cytidylyltransferase binding to membranes may be the ratio of bilayer to non-bilayer lipids in that membrane.

Full text

PDF
419

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Channon J. Y., Leslie C. C. A calcium-dependent mechanism for associating a soluble arachidonoyl-hydrolyzing phospholipase A2 with membrane in the macrophage cell line RAW 264.7. J Biol Chem. 1990 Apr 5;265(10):5409–5413. [PubMed] [Google Scholar]
  3. Choy P. C., Farren S. B., Vance D. E. Lipid requirements for the aggregation of CTP:phosphocholine cytidylyltransferase in rat liver cytosol. Can J Biochem. 1979 Jun;57(6):605–612. doi: 10.1139/o79-076. [DOI] [PubMed] [Google Scholar]
  4. Cornell R. B. Regulation of CTP:phosphocholine cytidylyltransferase by lipids. 1. Negative surface charge dependence for activation. Biochemistry. 1991 Jun 18;30(24):5873–5880. doi: 10.1021/bi00238a010. [DOI] [PubMed] [Google Scholar]
  5. Cornell R. B. Regulation of CTP:phosphocholine cytidylyltransferase by lipids. 2. Surface curvature, acyl chain length, and lipid-phase dependence for activation. Biochemistry. 1991 Jun 18;30(24):5881–5888. doi: 10.1021/bi00238a011. [DOI] [PubMed] [Google Scholar]
  6. Cornell R., Vance D. E. Translocation of CTP: phosphocholine cytidylyltransferase from cytosol to membranes in HeLa cells: stimulation by fatty acid, fatty alcohol, mono- and diacylglycerol. Biochim Biophys Acta. 1987 May 13;919(1):26–36. doi: 10.1016/0005-2760(87)90214-1. [DOI] [PubMed] [Google Scholar]
  7. Davis R. A., Engelhorn S. C., Pangburn S. H., Weinstein D. B., Steinberg D. Very low density lipoprotein synthesis and secretion by cultured rat hepatocytes. J Biol Chem. 1979 Mar 25;254(6):2010–2016. [PubMed] [Google Scholar]
  8. Feldman D. A., Brubaker P. G., Weinhold P. A. Activation of CTP : phosphocholine cytidylyltransferase in rat lung by fatty acids. Biochim Biophys Acta. 1981 Jul 24;665(1):53–59. doi: 10.1016/0005-2760(81)90231-9. [DOI] [PubMed] [Google Scholar]
  9. Feldman D. A., Weinhold P. A. CTP:phosphorylcholine cytidylyltransferase from rat liver. Isolation and characterization of the catalytic subunit. J Biol Chem. 1987 Jul 5;262(19):9075–9081. [PubMed] [Google Scholar]
  10. Gagné J., Stamatatos L., Diacovo T., Hui S. W., Yeagle P. L., Silvius J. R. Physical properties and surface interactions of bilayer membranes containing N-methylated phosphatidylethanolamines. Biochemistry. 1985 Jul 30;24(16):4400–4408. doi: 10.1021/bi00337a022. [DOI] [PubMed] [Google Scholar]
  11. Jamil H., Utal A. K., Vance D. E. Evidence that cyclic AMP-induced inhibition of phosphatidylcholine biosynthesis is caused by a decrease in cellular diacylglycerol levels in cultured rat hepatocytes. J Biol Chem. 1992 Jan 25;267(3):1752–1760. [PubMed] [Google Scholar]
  12. Jamil H., Vance D. E. Head-group specificity for feedback regulation of CTP:phosphocholine cytidylyltransferase. Biochem J. 1990 Sep 15;270(3):749–754. doi: 10.1042/bj2700749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jamil H., Yao Z. M., Vance D. E. Feedback regulation of CTP:phosphocholine cytidylyltransferase translocation between cytosol and endoplasmic reticulum by phosphatidylcholine. J Biol Chem. 1990 Mar 15;265(8):4332–4339. [PubMed] [Google Scholar]
  14. Jensen J. W., Schutzbach J. S. Modulation of dolichyl-phosphomannose synthase activity by changes in the lipid environment of the enzyme. Biochemistry. 1988 Aug 23;27(17):6315–6320. doi: 10.1021/bi00417a017. [DOI] [PubMed] [Google Scholar]
  15. Kent C. Regulation of phosphatidylcholine biosynthesis. Prog Lipid Res. 1990;29(2):87–105. doi: 10.1016/0163-7827(90)90010-i. [DOI] [PubMed] [Google Scholar]
  16. Kolesnick R. N., Hemer M. R. Physiologic 1,2-diacylglycerol levels induce protein kinase C-independent translocation of a regulatory enzyme. J Biol Chem. 1990 Jul 5;265(19):10900–10904. [PubMed] [Google Scholar]
  17. Morand J. N., Kent C. Localization of the membrane-associated CTP:phosphocholine cytidylyltransferase in Chinese hamster ovary cells with an altered membrane composition. J Biol Chem. 1989 Aug 15;264(23):13785–13792. [PubMed] [Google Scholar]
  18. Pelech S. L., Cook H. W., Paddon H. B., Vance D. E. Membrane-bound CTP:phosphocholine cytidylyltransferase regulates the rate of phosphatidylcholine synthesis in HeLa cells treated with unsaturated fatty acids. Biochim Biophys Acta. 1984 Oct 4;795(3):433–440. doi: 10.1016/0005-2760(84)90169-3. [DOI] [PubMed] [Google Scholar]
  19. Pelech S. L., Pritchard P. H., Brindley D. N., Vance D. E. Fatty acids promote translocation of CTP:phosphocholine cytidylyltransferase to the endoplasmic reticulum and stimulate rat hepatic phosphatidylcholine synthesis. J Biol Chem. 1983 Jun 10;258(11):6782–6788. [PubMed] [Google Scholar]
  20. Pritchard P. H., Chiang P. K., Cantoni G. L., Vance D. E. Inhibition of phosphatidylethanolamine N-methylation by 3-deazaadenosine stimulates the synthesis of phosphatidylcholine via the CDP-choline pathway. J Biol Chem. 1982 Jun 10;257(11):6362–6367. [PubMed] [Google Scholar]
  21. Sanghera J. S., Vance D. E. Stimulation of CTP: phosphocholine cytidylyltransferase and phosphatidylcholine synthesis by incubation of rat hepatocytes with phospholipase A2. Biochim Biophys Acta. 1990 Feb 23;1042(3):380–385. doi: 10.1016/0005-2760(90)90168-w. [DOI] [PubMed] [Google Scholar]
  22. Sen A., Isac T. V., Hui S. W. Bilayer packing stress and defects in mixed dilinoleoylphosphatidylethanolamine and palmitoyloleoylphosphatidylcholine and their susceptibility to phospholipase A2. Biochemistry. 1991 May 7;30(18):4516–4521. doi: 10.1021/bi00232a021. [DOI] [PubMed] [Google Scholar]
  23. Slack B. E., Breu J., Wurtman R. J. Production of diacylglycerol by exogenous phospholipase C stimulates CTP:phosphocholine cytidylyltransferase activity and phosphatidylcholine synthesis in human neuroblastoma cells. J Biol Chem. 1991 Dec 25;266(36):24503–24508. [PubMed] [Google Scholar]
  24. Sleight R., Kent C. Regulation of phosphatidylcholine biosynthesis in cultured chick embryonic muscle treated with phospholipase C. J Biol Chem. 1980 Nov 25;255(22):10644–10650. [PubMed] [Google Scholar]
  25. Sleight R., Kent C. Regulation of phosphatidylcholine biosynthesis in mammalian cells. I. Effects of phospholipase C treatment on phosphatidylcholine metabolism in Chinese hamster ovary cells and LM mouse fibroblasts. J Biol Chem. 1983 Jan 25;258(2):824–830. [PubMed] [Google Scholar]
  26. Sleight R., Kent C. Regulation of phosphatidylcholine biosynthesis in mammalian cells. II. Effects of phospholipase C treatment on the activity and subcellular distribution of CTP:phosphocholine cytidylyltransferase in Chinese hamster ovary and LM cell lines. J Biol Chem. 1983 Jan 25;258(2):831–835. [PubMed] [Google Scholar]
  27. Sleight R., Kent C. Regulation of phosphatidylcholine biosynthesis in mammalian cells. III. Effects of alterations in the phospholipid compositions of Chinese hamster ovary and LM cells on the activity and distribution of CTP:phosphocholine cytidylyltransferase. J Biol Chem. 1983 Jan 25;258(2):836–839. [PubMed] [Google Scholar]
  28. Sohal P. S., Cornell R. B. Sphingosine inhibits the activity of rat liver CTP:phosphocholine cytidylyltransferase. J Biol Chem. 1990 Jul 15;265(20):11746–11750. [PubMed] [Google Scholar]
  29. Tercé F., Record M., Ribbes G., Chap H., Douste-Blazy L. Intracellular processing of cytidylyltransferase in Krebs II cells during stimulation of phosphatidylcholine synthesis. Evidence that a plasma membrane modification promotes enzyme translocation specifically to the endoplasmic reticulum. J Biol Chem. 1988 Mar 5;263(7):3142–3149. [PubMed] [Google Scholar]
  30. Tijburg L. B., Geelen M. J., van Golde L. M. Regulation of the biosynthesis of triacylglycerol, phosphatidylcholine and phosphatidylethanolamine in the liver. Biochim Biophys Acta. 1989 Jul 17;1004(1):1–19. doi: 10.1016/0005-2760(89)90206-3. [DOI] [PubMed] [Google Scholar]
  31. Utal A. K., Jamil H., Vance D. E. Diacylglycerol signals the translocation of CTP:choline-phosphate cytidylyltransferase in HeLa cells treated with 12-O-tetradecanoylphorbol-13-acetate. J Biol Chem. 1991 Dec 15;266(35):24084–24091. [PubMed] [Google Scholar]
  32. Vance D. E., Pelech S. D., Choy P. C. CTP: phosphocholine cytidylyltransferase from rat liver. Methods Enzymol. 1981;71(Pt 100):576–581. doi: 10.1016/0076-6879(81)71070-x. [DOI] [PubMed] [Google Scholar]
  33. Vance D. E., Trip E. M., Paddon H. B. Poliovirus increases phosphatidylcholine biosynthesis in HeLa cells by stimulation of the rate-limiting reaction catalyzed by CTP: phosphocholine cytidylyltransferase. J Biol Chem. 1980 Feb 10;255(3):1064–1069. [PubMed] [Google Scholar]
  34. Vance J. E., Nguyen T. M., Vance D. E. The biosynthesis of phosphatidylcholine by methylation of phosphatidylethanolamine derived from ethanolamine is not required for lipoprotein secretion by cultured rat hepatocytes. Biochim Biophys Acta. 1986 Feb 28;875(3):501–509. doi: 10.1016/0005-2760(86)90070-6. [DOI] [PubMed] [Google Scholar]
  35. Weinhold P. A., Rounsifer M. E., Williams S. E., Brubaker P. G., Feldman D. A. CTP:phosphorylcholine cytidylyltransferase in rat lung. The effect of free fatty acids on the translocation of activity between microsomes and cytosol. J Biol Chem. 1984 Aug 25;259(16):10315–10321. [PubMed] [Google Scholar]
  36. Wright T. M., Rangan L. A., Shin H. S., Raben D. M. Kinetic analysis of 1,2-diacylglycerol mass levels in cultured fibroblasts. Comparison of stimulation by alpha-thrombin and epidermal growth factor. J Biol Chem. 1988 Jul 5;263(19):9374–9380. [PubMed] [Google Scholar]
  37. Yao Z. M., Jamil H., Vance D. E. Choline deficiency causes translocation of CTP:phosphocholine cytidylyltransferase from cytosol to endoplasmic reticulum in rat liver. J Biol Chem. 1990 Mar 15;265(8):4326–4331. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES