Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1997 Jul 15;325(Pt 2):435–440. doi: 10.1042/bj3250435

Stimulation of DNA synthesis by natural ceramide 1-phosphate.

A Gomez-Muñoz 1, L M Frago 1, L Alvarez 1, I Varela-Nieto 1
PMCID: PMC1218579  PMID: 9230125

Abstract

We found that natural (long-chain) ceramide 1-phosphate can be dispersed into aqueous solution when dissolved in an appropriate mixture of methanol/dodecane (49:1, v/v). This solvent mixture facilitates the interaction of this phosphosphingolipid with cells. Under these conditions, incubation of EGFR T17 fibroblasts with natural ceramide 1-phosphate caused a potent stimulation of DNA synthesis. This effect was accompanied by an increase in the levels of proliferating-cell nuclear antigen. Concentrations of natural ceramide 1-phosphate that stimulated the synthesis of DNA did not inhibit adenylate cyclase activity, nor did they stimulate phospholipase D. Natural ceramide 1-phosphate did not alter the cellular phosphorylation state of tyrosine residues or of mitogen-activated protein kinase. Furthermore, natural ceramide 1-phosphate failed to induce the expression of the proto-oncogenes c-myc and c-fos. Both the stimulation of DNA synthesis and the induction of proliferating-cell nuclear antigen by natural ceramide 1-phosphate were inhibited by natural ceramides. This work suggests that the use of methanol and dodecane to deliver natural ceramide 1-phosphate to cells may be useful for elucidation of the biological function(s) and mechanism(s) of action of ceramide 1-phosphate.

Full Text

The Full Text of this article is available as a PDF (331.6 KB).

Selected References

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

  1. Auwerx J., Staels B., Sassone-Corsi P. Coupled and uncoupled induction of fos and jun transcription by different second messengers in cells of hematopoietic origin. Nucleic Acids Res. 1990 Jan 25;18(2):221–228. doi: 10.1093/nar/18.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berger A., Bittman R., Schmidt R. R., Spiegel S. Structural requirements of sphingosylphosphocholine and sphingosine-1-phosphate for stimulation of activator protein-1 activity. Mol Pharmacol. 1996 Sep;50(3):451–457. [PubMed] [Google Scholar]
  3. Boudker O., Futerman A. H. Detection and characterization of ceramide-1-phosphate phosphatase activity in rat liver plasma membrane. J Biol Chem. 1993 Oct 15;268(29):22150–22155. [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Daub H., Weiss F. U., Wallasch C., Ullrich A. Role of transactivation of the EGF receptor in signalling by G-protein-coupled receptors. Nature. 1996 Feb 8;379(6565):557–560. doi: 10.1038/379557a0. [DOI] [PubMed] [Google Scholar]
  6. Dietrich D. R. Toxicological and pathological applications of proliferating cell nuclear antigen (PCNA), a novel endogenous marker for cell proliferation. Crit Rev Toxicol. 1993;23(1):77–109. doi: 10.3109/10408449309104075. [DOI] [PubMed] [Google Scholar]
  7. Dobrowsky R. T., Hannun Y. A. Ceramide stimulates a cytosolic protein phosphatase. J Biol Chem. 1992 Mar 15;267(8):5048–5051. [PubMed] [Google Scholar]
  8. Dressler K. A., Kolesnick R. N. Ceramide 1-phosphate, a novel phospholipid in human leukemia (HL-60) cells. Synthesis via ceramide from sphingomyelin. J Biol Chem. 1990 Sep 5;265(25):14917–14921. [PubMed] [Google Scholar]
  9. Goldkorn T., Dressler K. A., Muindi J., Radin N. S., Mendelsohn J., Menaldino D., Liotta D., Kolesnick R. N. Ceramide stimulates epidermal growth factor receptor phosphorylation in A431 human epidermoid carcinoma cells. Evidence that ceramide may mediate sphingosine action. J Biol Chem. 1991 Aug 25;266(24):16092–16097. [PubMed] [Google Scholar]
  10. Gomez-Muñoz A., Martin A., O'Brien L., Brindley D. N. Cell-permeable ceramides inhibit the stimulation of DNA synthesis and phospholipase D activity by phosphatidate and lysophosphatidate in rat fibroblasts. J Biol Chem. 1994 Mar 25;269(12):8937–8943. [PubMed] [Google Scholar]
  11. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  12. Gómez-Muñoz A., Waggoner D. W., O'Brien L., Brindley D. N. Interaction of ceramides, sphingosine, and sphingosine 1-phosphate in regulating DNA synthesis and phospholipase D activity. J Biol Chem. 1995 Nov 3;270(44):26318–26325. doi: 10.1074/jbc.270.44.26318. [DOI] [PubMed] [Google Scholar]
  13. Hale A. J., Smith C. A., Sutherland L. C., Stoneman V. E., Longthorne V. L., Culhane A. C., Williams G. T. Apoptosis: molecular regulation of cell death. Eur J Biochem. 1996 Feb 15;236(1):1–26. doi: 10.1111/j.1432-1033.1996.00001.x. [DOI] [PubMed] [Google Scholar]
  14. Hannun Y. A., Obeid L. M., Wolff R. A. The novel second messenger ceramide: identification, mechanism of action, and cellular activity. Adv Lipid Res. 1993;25:43–64. [PubMed] [Google Scholar]
  15. Hannun Y. A. The sphingomyelin cycle and the second messenger function of ceramide. J Biol Chem. 1994 Feb 4;269(5):3125–3128. [PubMed] [Google Scholar]
  16. Hayakawa M., Jayadev S., Tsujimoto M., Hannun Y. A., Ito F. Role of ceramide in stimulation of the transcription of cytosolic phospholipase A2 and cyclooxygenase 2. Biochem Biophys Res Commun. 1996 Mar 27;220(3):681–686. doi: 10.1006/bbrc.1996.0464. [DOI] [PubMed] [Google Scholar]
  17. Ji L., Zhang G., Uematsu S., Akahori Y., Hirabayashi Y. Induction of apoptotic DNA fragmentation and cell death by natural ceramide. FEBS Lett. 1995 Jan 23;358(2):211–214. doi: 10.1016/0014-5793(94)01428-4. [DOI] [PubMed] [Google Scholar]
  18. Junco M., Díaz-Guerra M. J., Boscá L. Differential calcium mobilization by vasopressin, angiotensin II, gastrin-releasing peptide, and adenosine triphosphate in adult and fetal hepatocytes. Relevance for the activation of calcium-dependent enzymes. Endocrinology. 1993 Jan;132(1):309–318. doi: 10.1210/endo.132.1.8380381. [DOI] [PubMed] [Google Scholar]
  19. Kanety H., Hemi R., Papa M. Z., Karasik A. Sphingomyelinase and ceramide suppress insulin-induced tyrosine phosphorylation of the insulin receptor substrate-1. J Biol Chem. 1996 Apr 26;271(17):9895–9897. doi: 10.1074/jbc.271.17.9895. [DOI] [PubMed] [Google Scholar]
  20. Kim M. Y., Linardic C., Obeid L., Hannun Y. Identification of sphingomyelin turnover as an effector mechanism for the action of tumor necrosis factor alpha and gamma-interferon. Specific role in cell differentiation. J Biol Chem. 1991 Jan 5;266(1):484–489. [PubMed] [Google Scholar]
  21. Kolesnick R. N., Hemer M. R. Characterization of a ceramide kinase activity from human leukemia (HL-60) cells. Separation from diacylglycerol kinase activity. J Biol Chem. 1990 Nov 5;265(31):18803–18808. [PubMed] [Google Scholar]
  22. Kolesnick R., Golde D. W. The sphingomyelin pathway in tumor necrosis factor and interleukin-1 signaling. Cell. 1994 May 6;77(3):325–328. doi: 10.1016/0092-8674(94)90147-3. [DOI] [PubMed] [Google Scholar]
  23. Liscovitch M., Chalifa V., Danin M., Eli Y. Inhibition of neural phospholipase D activity by aminoglycoside antibiotics. Biochem J. 1991 Oct 1;279(Pt 1):319–321. doi: 10.1042/bj2790319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Martin A., Gomez-Muñoz A., Waggoner D. W., Stone J. C., Brindley D. N. Decreased activities of phosphatidate phosphohydrolase and phospholipase D in ras and tyrosine kinase (fps) transformed fibroblasts. J Biol Chem. 1993 Nov 15;268(32):23924–23932. [PubMed] [Google Scholar]
  25. Mathias S., Dressler K. A., Kolesnick R. N. Characterization of a ceramide-activated protein kinase: stimulation by tumor necrosis factor alpha. Proc Natl Acad Sci U S A. 1991 Nov 15;88(22):10009–10013. doi: 10.1073/pnas.88.22.10009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Merrill A. H., Jr, Stevens V. L. Modulation of protein kinase C and diverse cell functions by sphingosine--a pharmacologically interesting compound linking sphingolipids and signal transduction. Biochim Biophys Acta. 1989 Feb 9;1010(2):131–139. doi: 10.1016/0167-4889(89)90152-3. [DOI] [PubMed] [Google Scholar]
  27. Moolenaar W. H., Kruijer W., Tilly B. C., Verlaan I., Bierman A. J., de Laat S. W. Growth factor-like action of phosphatidic acid. Nature. 1986 Sep 11;323(6084):171–173. doi: 10.1038/323171a0. [DOI] [PubMed] [Google Scholar]
  28. Obeid L. M., Linardic C. M., Karolak L. A., Hannun Y. A. Programmed cell death induced by ceramide. Science. 1993 Mar 19;259(5102):1769–1771. doi: 10.1126/science.8456305. [DOI] [PubMed] [Google Scholar]
  29. Okazaki T., Bielawska A., Bell R. M., Hannun Y. A. Role of ceramide as a lipid mediator of 1 alpha,25-dihydroxyvitamin D3-induced HL-60 cell differentiation. J Biol Chem. 1990 Sep 15;265(26):15823–15831. [PubMed] [Google Scholar]
  30. Pagano R. E., Sleight R. G. Defining lipid transport pathways in animal cells. Science. 1985 Sep 13;229(4718):1051–1057. doi: 10.1126/science.4035344. [DOI] [PubMed] [Google Scholar]
  31. Preiss J., Loomis C. R., Bishop W. R., Stein R., Niedel J. E., Bell R. M. Quantitative measurement of sn-1,2-diacylglycerols present in platelets, hepatocytes, and ras- and sis-transformed normal rat kidney cells. J Biol Chem. 1986 Jul 5;261(19):8597–8600. [PubMed] [Google Scholar]
  32. Raines M. A., Kolesnick R. N., Golde D. W. Sphingomyelinase and ceramide activate mitogen-activated protein kinase in myeloid HL-60 cells. J Biol Chem. 1993 Jul 15;268(20):14572–14575. [PubMed] [Google Scholar]
  33. Schneider E. G., Kennedy E. P. Phosphorylation of ceramide by diglyceride kinase preparations from Escherichia coli. J Biol Chem. 1973 May 25;248(10):3739–3741. [PubMed] [Google Scholar]
  34. Shinghal R., Scheller R. H., Bajjalieh S. M. Ceramide 1-phosphate phosphatase activity in brain. J Neurochem. 1993 Dec;61(6):2279–2285. doi: 10.1111/j.1471-4159.1993.tb07470.x. [DOI] [PubMed] [Google Scholar]
  35. Stanton L. W., Watt R., Marcu K. B. Translocation, breakage and truncated transcripts of c-myc oncogene in murine plasmacytomas. Nature. 1983 Jun 2;303(5916):401–406. doi: 10.1038/303401a0. [DOI] [PubMed] [Google Scholar]
  36. Su Y., Rosenthal D., Smulson M., Spiegel S. Sphingosine 1-phosphate, a novel signaling molecule, stimulates DNA binding activity of AP-1 in quiescent Swiss 3T3 fibroblasts. J Biol Chem. 1994 Jun 10;269(23):16512–16517. [PubMed] [Google Scholar]
  37. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Tovey K. C., Oldham K. G., Whelan J. A. A simple direct assay for cyclic AMP in plasma and other biological samples using an improved competitive protein binding technique. Clin Chim Acta. 1974 Nov 8;56(3):221–234. doi: 10.1016/0009-8981(74)90133-8. [DOI] [PubMed] [Google Scholar]
  39. Velu T. J., Beguinot L., Vass W. C., Willingham M. C., Merlino G. T., Pastan I., Lowy D. R. Epidermal-growth-factor-dependent transformation by a human EGF receptor proto-oncogene. Science. 1987 Dec 4;238(4832):1408–1410. doi: 10.1126/science.3500513. [DOI] [PubMed] [Google Scholar]
  40. Verheij M., Bose R., Lin X. H., Yao B., Jarvis W. D., Grant S., Birrer M. J., Szabo E., Zon L. I., Kyriakis J. M. Requirement for ceramide-initiated SAPK/JNK signalling in stress-induced apoptosis. Nature. 1996 Mar 7;380(6569):75–79. doi: 10.1038/380075a0. [DOI] [PubMed] [Google Scholar]
  41. Waggoner D. W., Gómez-Muñoz A., Dewald J., Brindley D. N. Phosphatidate phosphohydrolase catalyzes the hydrolysis of ceramide 1-phosphate, lysophosphatidate, and sphingosine 1-phosphate. J Biol Chem. 1996 Jul 12;271(28):16506–16509. doi: 10.1074/jbc.271.28.16506. [DOI] [PubMed] [Google Scholar]
  42. Zhang H., Buckley N. E., Gibson K., Spiegel S. Sphingosine stimulates cellular proliferation via a protein kinase C-independent pathway. J Biol Chem. 1990 Jan 5;265(1):76–81. [PubMed] [Google Scholar]
  43. Zhang H., Desai N. N., Olivera A., Seki T., Brooker G., Spiegel S. Sphingosine-1-phosphate, a novel lipid, involved in cellular proliferation. J Cell Biol. 1991 Jul;114(1):155–167. doi: 10.1083/jcb.114.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. van Corven E. J., Groenink A., Jalink K., Eichholtz T., Moolenaar W. H. Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. Cell. 1989 Oct 6;59(1):45–54. doi: 10.1016/0092-8674(89)90868-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES