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. 1998 Mar 1;330(Pt 2):1009–1014. doi: 10.1042/bj3301009

Exogenous C2-ceramide activates c-fos serum response element via Rac-dependent signalling pathway.

B C Kim 1, J H Kim 1
PMCID: PMC1219238  PMID: 9480923

Abstract

Ceramide is an important regulatory molecule implicated in a variety of biological processes in response to stress and cytokines. To understand the signal transduction pathway of ceramide to the nucleus, in the present study, we examined whether C2-ceramide, a cell permeable ceramide, activates c-fos serum response element (SRE). Treatment of Rat-2 fibroblast cells with C2-ceramide caused the stimulation of c-fos SRE-dependent reporter gene activity in a dose- and time-dependent manner by transient transfection analysis. Next, we examined the role of Rho family GTPases in the ceramide-induced signalling to SRE activation. By reporter gene analysis following transient transfections with various plasmids expressing a dominant negative mutant form of Cdc42, Rac1 or RhoA, C2-ceramide-induced SRE activation was shown to be selectively repressed by pEXV-RacN17 encoding a dominant negative mutant of Rac1, suggesting that Rac activity is essential for the signalling cascade of ceramide to the nucleus. In a further study to analyse the downstream mediator of Rac in the ceramide-signalling pathway, we observed that either pretreatment with mepacrine, a potent and specific inhibitor of phospholipase A2, or co-transfection with antisense cytosolic phospholipase A2 (cPLA2) oligonucleotide repressed the C2-ceramide-induced SRE activation selectively, implying a critical role of cPLA2 in C2-ceramide-induced signalling to nucleus. Consistent with these results, the translocation of cPLA2 protein as well as the release of arachidonic acid, a principal product of phospholipase A2, was rapidly induced by the addition of C2-ceramide in a Rac-dependent manner. Together, our findings suggest the critical role of 'Rac and subsequent activation of phospholipase A2' in ceramide-signalling to nucleus.

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Selected References

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  1. Bagrodia S., Dérijard B., Davis R. J., Cerione R. A. Cdc42 and PAK-mediated signaling leads to Jun kinase and p38 mitogen-activated protein kinase activation. J Biol Chem. 1995 Nov 24;270(47):27995–27998. doi: 10.1074/jbc.270.47.27995. [DOI] [PubMed] [Google Scholar]
  2. Ballou L. R., Laulederkind S. J., Rosloniec E. F., Raghow R. Ceramide signalling and the immune response. Biochim Biophys Acta. 1996 Jun 11;1301(3):273–287. doi: 10.1016/0005-2760(96)00004-5. [DOI] [PubMed] [Google Scholar]
  3. Cavigelli M., Dolfi F., Claret F. X., Karin M. Induction of c-fos expression through JNK-mediated TCF/Elk-1 phosphorylation. EMBO J. 1995 Dec 1;14(23):5957–5964. doi: 10.1002/j.1460-2075.1995.tb00284.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou M. M., Blenis J. The 70 kDa S6 kinase complexes with and is activated by the Rho family G proteins Cdc42 and Rac1. Cell. 1996 May 17;85(4):573–583. doi: 10.1016/s0092-8674(00)81257-x. [DOI] [PubMed] [Google Scholar]
  5. Coso O. A., Chiariello M., Yu J. C., Teramoto H., Crespo P., Xu N., Miki T., Gutkind J. S. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway. Cell. 1995 Jun 30;81(7):1137–1146. doi: 10.1016/s0092-8674(05)80018-2. [DOI] [PubMed] [Google Scholar]
  6. Cuvillier O., Pirianov G., Kleuser B., Vanek P. G., Coso O. A., Gutkind S., Spiegel S. Suppression of ceramide-mediated programmed cell death by sphingosine-1-phosphate. Nature. 1996 Jun 27;381(6585):800–803. doi: 10.1038/381800a0. [DOI] [PubMed] [Google Scholar]
  7. Gille H., Strahl T., Shaw P. E. Activation of ternary complex factor Elk-1 by stress-activated protein kinases. Curr Biol. 1995 Oct 1;5(10):1191–1200. doi: 10.1016/s0960-9822(95)00235-1. [DOI] [PubMed] [Google Scholar]
  8. Hall A. Small GTP-binding proteins and the regulation of the actin cytoskeleton. Annu Rev Cell Biol. 1994;10:31–54. doi: 10.1146/annurev.cb.10.110194.000335. [DOI] [PubMed] [Google Scholar]
  9. Hannun Y. A. Functions of ceramide in coordinating cellular responses to stress. Science. 1996 Dec 13;274(5294):1855–1859. doi: 10.1126/science.274.5294.1855. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Hill C. S., Wynne J., Treisman R. The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF. Cell. 1995 Jun 30;81(7):1159–1170. doi: 10.1016/s0092-8674(05)80020-0. [DOI] [PubMed] [Google Scholar]
  13. Jarvis W. D., Grant S., Kolesnick R. N. Ceramide and the induction of apoptosis. Clin Cancer Res. 1996 Jan;2(1):1–6. [PubMed] [Google Scholar]
  14. Jayadev S., Linardic C. M., Hannun Y. A. Identification of arachidonic acid as a mediator of sphingomyelin hydrolysis in response to tumor necrosis factor alpha. J Biol Chem. 1994 Feb 25;269(8):5757–5763. [PubMed] [Google Scholar]
  15. Johansen F. E., Prywes R. Two pathways for serum regulation of the c-fos serum response element require specific sequence elements and a minimal domain of serum response factor. Mol Cell Biol. 1994 Sep;14(9):5920–5928. doi: 10.1128/mcb.14.9.5920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jättelä M., Benedict M., Tewari M., Shayman J. A., Dixit V. M. Bcl-x and Bcl-2 inhibit TNF and Fas-induced apoptosis and activation of phospholipase A2 in breast carcinoma cells. Oncogene. 1995 Jun 15;10(12):2297–2305. [PubMed] [Google Scholar]
  17. Kim B. C., Kim J. H. Role of Rac GTPase in the nuclear signaling by EGF. FEBS Lett. 1997 Apr 21;407(1):7–12. doi: 10.1016/s0014-5793(97)00289-5. [DOI] [PubMed] [Google Scholar]
  18. Kim J. H., Johansen F. E., Robertson N., Catino J. J., Prywes R., Kumar C. C. Suppression of Ras transformation by serum response factor. J Biol Chem. 1994 May 13;269(19):13740–13743. [PubMed] [Google Scholar]
  19. Kim J. H., Kwack H. J., Choi S. E., Kim B. C., Kim Y. S., Kang I. J., Kumar C. C. Essential role of Rac GTPase in hydrogen peroxide-induced activation of c-fos serum response element. FEBS Lett. 1997 Apr 7;406(1-2):93–96. doi: 10.1016/s0014-5793(97)00249-4. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Manser E., Leung T., Salihuddin H., Zhao Z. S., Lim L. A brain serine/threonine protein kinase activated by Cdc42 and Rac1. Nature. 1994 Jan 6;367(6458):40–46. doi: 10.1038/367040a0. [DOI] [PubMed] [Google Scholar]
  22. Minden A., Lin A., Claret F. X., Abo A., Karin M. Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs. Cell. 1995 Jun 30;81(7):1147–1157. doi: 10.1016/s0092-8674(05)80019-4. [DOI] [PubMed] [Google Scholar]
  23. Peppelenbosch M. P., Tertoolen L. G., Hage W. J., de Laat S. W. Epidermal growth factor-induced actin remodeling is regulated by 5-lipoxygenase and cyclooxygenase products. Cell. 1993 Aug 13;74(3):565–575. doi: 10.1016/0092-8674(93)80057-l. [DOI] [PubMed] [Google Scholar]
  24. Peppelenbosch M. P., Tertoolen L. G., de Vries-Smits A. M., Qiu R. G., M'Rabet L., Symons M. H., de Laat S. W., Bos J. L. Rac-dependent and -independent pathways mediate growth factor-induced Ca2+ influx. J Biol Chem. 1996 Apr 5;271(14):7883–7886. doi: 10.1074/jbc.271.14.7883. [DOI] [PubMed] [Google Scholar]
  25. Peppelenbosch M. P., Tertoolen L. G., den Hertog J., de Laat S. W. Epidermal growth factor activates calcium channels by phospholipase A2/5-lipoxygenase-mediated leukotriene C4 production. Cell. 1992 Apr 17;69(2):295–303. doi: 10.1016/0092-8674(92)90410-e. [DOI] [PubMed] [Google Scholar]
  26. Spiegel S., Foster D., Kolesnick R. Signal transduction through lipid second messengers. Curr Opin Cell Biol. 1996 Apr;8(2):159–167. doi: 10.1016/s0955-0674(96)80061-5. [DOI] [PubMed] [Google Scholar]
  27. Tang D. G., Chen Y. Q., Honn K. V. Arachidonate lipoxygenases as essential regulators of cell survival and apoptosis. Proc Natl Acad Sci U S A. 1996 May 28;93(11):5241–5246. doi: 10.1073/pnas.93.11.5241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Treisman R. The SRE: a growth factor responsive transcriptional regulator. Semin Cancer Biol. 1990 Feb;1(1):47–58. [PubMed] [Google Scholar]
  29. 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]
  30. Visnjić D., Batinić D., Banfić H. Arachidonic acid mediates interferon-gamma-induced sphingomyelin hydrolysis and monocytic marker expression in HL-60 cell line. Blood. 1997 Jan 1;89(1):81–91. [PubMed] [Google Scholar]
  31. Whitmarsh A. J., Shore P., Sharrocks A. D., Davis R. J. Integration of MAP kinase signal transduction pathways at the serum response element. Science. 1995 Jul 21;269(5222):403–407. doi: 10.1126/science.7618106. [DOI] [PubMed] [Google Scholar]

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