Key points are not available for this paper at this time.
Ceramide, a sphingolipid generated by the hydrolysis of membrane-associated sphingomyelin, appears to play a role as a gauge of apoptosis. A further metabolite of ceramide, sphingosine 1-phosphate (SPP), prevents ceramide-mediated apoptosis, and it has been suggested that the balance between intracellular ceramide and SPP levels may determine the cell fate (Cuvillier, O., Pirianov, G, Kleuser, B., Vanek, P. G., Coso, O. A., Gutkind, J. S., and Spiegel, S. (1996) Nature 381, 800–803). Here, we investigated the role of SPP and the protein kinase C activator, phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), in the caspase cascade leading to the proteolysis of poly(ADP-ribose) polymerase (PARP) and lamins. In Jurkat T cells, Fas ligation or addition of exogenous C2-ceramide induced activations of caspase-3/CPP32 and caspase-7/Mch3 followed by PARP cleavage, effects that can be blocked either by SPP or TPA. Furthermore, both SPP and TPA inhibit the activation of caspase-6/Mch2 and subsequent lamin B cleavage. Ceramide, in contrast to Fas ligation, did not induce activation of caspase-8/FLICE and neither SPP nor TPA were able to prevent this activation. Thus, SPP, likely generated via protein kinase C-mediated activation of sphingosine kinase, suppresses the apoptotic pathway downstream of FLICE but upstream of the executioner caspases, caspase-3, -6, and -7. Ceramide, a sphingolipid generated by the hydrolysis of membrane-associated sphingomyelin, appears to play a role as a gauge of apoptosis. A further metabolite of ceramide, sphingosine 1-phosphate (SPP), prevents ceramide-mediated apoptosis, and it has been suggested that the balance between intracellular ceramide and SPP levels may determine the cell fate (Cuvillier, O., Pirianov, G, Kleuser, B., Vanek, P. G., Coso, O. A., Gutkind, J. S., and Spiegel, S. (1996) Nature 381, 800–803). Here, we investigated the role of SPP and the protein kinase C activator, phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), in the caspase cascade leading to the proteolysis of poly(ADP-ribose) polymerase (PARP) and lamins. In Jurkat T cells, Fas ligation or addition of exogenous C2-ceramide induced activations of caspase-3/CPP32 and caspase-7/Mch3 followed by PARP cleavage, effects that can be blocked either by SPP or TPA. Furthermore, both SPP and TPA inhibit the activation of caspase-6/Mch2 and subsequent lamin B cleavage. Ceramide, in contrast to Fas ligation, did not induce activation of caspase-8/FLICE and neither SPP nor TPA were able to prevent this activation. Thus, SPP, likely generated via protein kinase C-mediated activation of sphingosine kinase, suppresses the apoptotic pathway downstream of FLICE but upstream of the executioner caspases, caspase-3, -6, and -7. Physiologic cell death occurs through an evolutionary conserved suicide process, termed apoptosis, which plays a considerable role in early development and homeostasis of adult tissues (1Vaux D.L. Strasser A. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: 2239-2244Crossref PubMed Scopus (913) Google Scholar). Ceramide has recently emerged as a critical component of apoptosis (2Obeid L.M. Linardic C.M. Karolak L.A. Hannun Y.A. Science. 1993; 259: 1769-1771Crossref PubMed Scopus (1631) Google Scholar, 3Hannun Y. Science. 1996; 274: 1855-1859Crossref PubMed Scopus (1510) Google Scholar). A variety of stress stimuli, such as tumor necrosis factor α (TNFα), 1The abbreviations used are: TNFα, tumor necrosis factor α; Ac-DEVD-AMC, acetyl-Asp-Glu-Val-Asp-aminomethylcoumarin; DEVD, Asp-Glu-Val-Asp; ICE, interleukin1β-converting enzyme; PARP, enzyme poly(ADP-ribose) polymerase; PKC, protein kinase C; SPP, sphingosine 1-phosphate; TPA, 12-O-tetradecanoylphorbol-13-acetate; PBS, phsophate-buffered saline; JNK, c-Jun N-terminal kinase; SAPK, stress-activated protein kinase; CHAPS, 3-(3-cholamidopropyl)dimethylammonio-1-propanesulfonic acid. Fas ligand, growth factor withdrawal, anticancer drugs, oxidative stress, heat shock, ionizing radiation, and ultraviolet light increase cellular ceramide which, in turn, is capable of inducing apoptosis (3Hannun Y. Science. 1996; 274: 1855-1859Crossref PubMed Scopus (1510) Google Scholar, 4Spiegel S. Foster D. Kolesnick R. Curr. Opin. Cell Biol. 1996; 8: 159-167Crossref PubMed Scopus (475) Google Scholar). This apoptotic effect is blocked by addition of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) or diacylglycerol, both activators of protein kinase C (PKC) (2Obeid L.M. Linardic C.M. Karolak L.A. Hannun Y.A. Science. 1993; 259: 1769-1771Crossref PubMed Scopus (1631) Google Scholar, 5Jarvis W.D. Fornari Jr., F.A. Browning J.L. Gewirtz D.A. Kolesnick R.N. Grant S. J. Biol. Chem. 1994; 269: 31685-31692Abstract Full Text PDF PubMed Google Scholar), suggesting that PKC activation counteracts ceramide-mediated apoptosis. Activation of PKC in diverse cell types stimulates sphingosine kinase activity resulting in intracellular accumulation of sphingosine 1-phosphate (SPP) (6Mazurek N. Megidish T. Hakomori S.-I. Igarashi Y. Biochem. Biophys. Res. Commun. 1994; 198: 1-9Crossref PubMed Scopus (89) Google Scholar, 7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). Recently, we showed that SPP prevented the hallmarks of apoptosis resulting from elevated levels of ceramide induced by TNFα, anti-Fas antibody, sphingomyelinase, or cell-permeable ceramide (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). Furthermore, inhibition of PKC, as well as inhibition of sphingosine kinase, induces apoptosis, which can be overcome by the addition of SPP (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). These results indicate that PKC may inhibit ceramide-induced apoptosis by activating sphingosine kinase. Recently, attention has been focused on the role of a novel family of aspartate-specific cysteine proteases, called caspases, which are intimately associated with apoptosis (reviewed in Ref. 8Nicholson D.W. Thornberry N.A. Trends Biochem. Sci. 1997; 22: 299-306Abstract Full Text PDF PubMed Scopus (2201) Google Scholar). Genetic studies of the nematode Caenorhabditis elegans have led to the identification of two genes, ced-3 and ced-4, that are required for apoptotic cell death (9Yuan J. Shaham S. Ledoux S. Ellis H.M. Horvitz H.R. Cell. 1993; 75: 641-652Abstract Full Text PDF PubMed Scopus (2268) Google Scholar). The ced-3gene encodes a caspase similar to the prototype mammalian interleukin 1β-converting enzyme (ICE) (9Yuan J. Shaham S. Ledoux S. Ellis H.M. Horvitz H.R. Cell. 1993; 75: 641-652Abstract Full Text PDF PubMed Scopus (2268) Google Scholar). Ten other homologs of ICE/CED-3 have been identified and phylogenetically classified into three subfamilies: (i) the CED-3 subfamily, consisting of caspase-3 (CPP32/Yama/apopain), caspase-6 (Mch2), caspase-7 (Mch3/ICE-LAP3/CMH-1), caspase-8 (FLICE/MACH/Mch5), caspase-9 (ICE-LAP6/Mch6), and caspase-10 (Mch4); (ii) the ICE subfamily, which includes caspase-1 (ICE), caspase-4 (TX/ICH2/ICE rel-II), caspase-5 (TY/ICE rel-III), and caspase-11 (ICH3); and (iii) the Nedd-2 subfamily, which is represented by caspase-2 (ICH1/murine Nedd-2). Previous studies have identified a number of substrates for the caspases, particularly the DNA repair enzyme poly(ADP-ribose) polymerase (PARP) (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar) and the nuclear lamins (11Lazebnik Y.A. Takahashi A. Moir R.D. Goldman R.D. Poirier G.G. Kaufman S.H. Earnshaw W.C. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 9042-9046Crossref PubMed Scopus (484) Google Scholar). Proteolysis of these substrates may account for many of the biochemical and morphological nuclear changes associated with apoptosis. The early of PARP can be by to the CED-3 (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar, M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar, M. J. Biol. Chem. 1996; PubMed Scopus Google Scholar). caspase-3/CPP32 is to be the PARP (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar, M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar), followed by caspase-7/Mch3 T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar). The of nuclear lamins is required for of the into apoptotic a the of apoptosis (11Lazebnik Y.A. Takahashi A. Moir R.D. Goldman R.D. Poirier G.G. Kaufman S.H. Earnshaw W.C. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 9042-9046Crossref PubMed Scopus (484) Google Scholar). is the caspase capable of lamins M. J. Biol. Chem. 1996; PubMed Scopus Google Scholar, A. Lazebnik Y.A. T. G. Moir R.D. Goldman R.D. Poirier G.G. S.H. Earnshaw W.C. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: PubMed Scopus Google Scholar). the intracellular levels of ceramide and SPP have been to be a critical gauge of cell fate (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar), the of of these sphingolipid are not well Recently, ceramide has been in PARP D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar, D.K. J. Poirier G.G. Hannun Y.A. L.M. Biochem. J. 1996; PubMed Scopus Google Scholar) and caspase-3/CPP32 activation N. R. Y. S. N. 1996; PubMed Scopus Google Scholar). In it appears that ceramide can be generated via a likely of both cell death and ceramide accumulation induced by TNFα, exogenous ceramide can this and induce apoptosis D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar). In this the between sphingolipid and the that both SPP and TPA PARP by activation of caspase-3/CPP32 and caspase-7/Mch3 induced by Fas ligation or cell-permeable Furthermore, we that SPP and TPA inhibit activation of caspase-6/Mch2 and subsequent lamin B cleavage. ceramide, in contrast to Fas ligation, did not induce activation of caspase-8/FLICE and neither SPP nor TPA were able to prevent this activation. from C2-ceramide and SPP were from from of from from and and to were from Jurkat T were in with the of the were in and in of and in this DNA in with for in Scholar). were in in and to the were in and for on DNA and were by for were in and to a of DNA as DNA All results were in and as by with in for were with a A for PARP of used to the of PARP with by in a by on a CHAPS, and were by cell in CHAPS, and PARP activity by of protein from Jurkat T cells, CHAPS, and in a of were for and by the addition of of and for were by were in for with for and protein by were on a and the and PARP were by in for on and to blocked by of the with for were for with for the of caspase-3/CPP32 of for the of caspase-6/Mch2 B for the of or for the of caspase-8/FLICE of with were for with three with used to the were and by were in and of in of with of A a an of and a of were used as (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar, N. R. Y. S. N. 1996; PubMed Scopus Google Scholar, R. P. M. A. R. J. 1994; PubMed Scopus Google Scholar), Jurkat with Fas cell death as by the DNA of cell and were into and apoptotic In with (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar), were with TPA, DNA with SPP apoptosis by and Ceramide has been to play a role in the Fas pathway R. P. M. A. R. J. 1994; PubMed Scopus Google Scholar, S. M.E. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). of Jurkat T with the cell-permeable C2-ceramide in cell death that overcome either by with TPA or to a by SPP of sphingosine kinase leading to intracellular accumulation of SPP has been to be of the effects by activation of PKC (6Mazurek N. Megidish T. Hakomori S.-I. Igarashi Y. Biochem. Biophys. Res. Commun. 1994; 198: 1-9Crossref PubMed Scopus (89) Google Scholar, 7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). activation of PKC by phorbol ceramide (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar, A. D. M. Kolesnick R.N. J. 1994; PubMed Scopus Google Scholar). In the PKC and induce hydrolysis to ceramide J. Res. 1996; Google Scholar, D.A. G. J. 1996; PubMed Scopus Google Scholar). These studies that PKC activation may the Fas apoptotic both by activation of sphingosine kinase and by the levels of Fas ligation to of PARP M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, Poirier G.G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, Miller D.K. Yamin T. A. 1996; PubMed Scopus Google Scholar, J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar), it of to determine SPP and TPA inhibit this cleavage. In with results Poirier G.G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, Miller D.K. Yamin T. A. 1996; PubMed Scopus Google Scholar, J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar), from to anti-Fas for were able to in into the apoptotic that of the PARP to the by from SPP of PARP induced by Fas ligation by and as with TPA prevented PARP Ceramide has recently been to PARP in apoptosis D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar). In of or with exogenous results in of PARP D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar, D.K. J. Poirier G.G. Hannun Y.A. L.M. Biochem. J. 1996; PubMed Scopus Google Scholar), suggesting that ceramide a caspase for PARP cleavage. In with these studies D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar, D.K. J. Poirier G.G. Hannun Y.A. L.M. Biochem. J. 1996; PubMed Scopus Google Scholar), we that C2-ceramide induced PARP and with TPA prevented this SPP PARP proteolysis by These results indicate that SPP can inhibit PARP induced either by Fas ligation or exogenous is as the with a (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar, M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar). which the to can PARP, a T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar). used the Ac-DEVD-AMC, which to the that is in PARP (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar), to the activity of these from to anti-Fas showed a increase in activity as N. R. Y. S. N. 1996; PubMed Scopus Google Scholar, J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). In with TPA blocked this SPP caspase activity of in to Fas by for the This caspase is as a that is to the of through an and (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar, M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, T. G. J. Biol. Chem. 1994; 269: Full Text PDF PubMed Google Scholar). In with studies Poirier G.G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar), Fas the into caspase-3 blocked by TPA, as not in with both anti-Fas and TPA. SPP prevented caspase-3/CPP32 by as by A similar used to the effects of SPP and TPA on exogenous caspase-3/CPP32 activation. In with a N. R. Y. S. N. 1996; PubMed Scopus Google Scholar), from with C2-ceramide an increase in activity with the of the and of caspase-3 TPA blocked both the activity increase and the of caspase-3/CPP32 into and by ceramide from with SPP activity the inhibition of and which is capable of PARP T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar), is as a and activation is into and Poirier G.G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, J.P. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). Fas ligation to the of the and Poirier G.G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, J.P. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). Thus, it of to ceramide capable of inducing activation of and SPP this Fas induced the of the which by with TPA and to a with SPP of to the induced by ceramide by with TPA, and SPP by Thus, SPP is able to of caspase-3 and and subsequent of PARP by Fas or exogenous In addition to the of the nuclear enzyme PARP, that is to apoptosis is the of which play in nuclear (11Lazebnik Y.A. Takahashi A. Moir R.D. Goldman R.D. Poirier G.G. Kaufman S.H. Earnshaw W.C. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 9042-9046Crossref PubMed Scopus (484) Google Scholar). caspase-6/Mch2 is the M. J. Biol. Chem. 1996; PubMed Scopus Google Scholar, A. Lazebnik Y.A. T. G. Moir R.D. Goldman R.D. Poirier G.G. S.H. Earnshaw W.C. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: PubMed Scopus Google Scholar). of Fas in Jurkat T has been to caspase-6 M. J. Biol. Chem. 1996; PubMed Scopus Google Scholar) and the of lamin B Miller D.K. Yamin T. A. 1996; PubMed Scopus Google Scholar). Thus, we ceramide induce caspase-6 activation and lamin B cleavage. B is into a with Fas or exogenous C2-ceramide A and of from with TPA and Fas or C2-ceramide lamin of inhibition of cleavage. were with SPP or apoptosis lamin B proteolysis caspase-6 by SPP and TPA from activation by and C2-ceramide In with studies M. J. Biol. Chem. 1996; PubMed Scopus Google Scholar), Fas able to caspase-6 cleavage, and with C2-ceramide led to of caspase-6 were with TPA or SPP, of of and A and SPP, TPA, can of caspase-6 and nuclear or of induced by Fas or C2-ceramide is by SPP and TPA. Cell from Jurkat for in with Fas or C2-ceramide in the or of TPA or the of SPP, were by to and with for the of The indicate the of the and and ceramide accumulation induced by or Fas are by a of the exogenous ceramide is able to this and induce apoptosis by activating downstream D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar). is a of ICE P.R. Cell. Full Text PDF PubMed Scopus Google Scholar). the of in apoptosis is M. S. Nature. 1995; PubMed Scopus Google and M. M. M. Nature. 1995; PubMed Scopus Google J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google and J. S. S. T. Y. Res. 1996; Google Scholar). the upstream caspase in and apoptosis, appears to be the of in S. M. J. Biol. Chem. 1997; PubMed Scopus Google Scholar). Thus, it of to determine the of sphingolipid in activation of induced the of a of and to the of FLICE In C2-ceramide did not induce of caspase-8 This is in with the that ceramide downstream of a neither SPP nor TPA have an effect on caspase-8/FLICE the studies have been which have of cell are to the cell suicide S. M.E. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar, M. A. J. M. R. M. M.E. Cell. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). death of cell for or Fas to a cascade of of the protein which a death either to or to via death protein J. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar), caspase-8 to the resulting in activation of the M. A. J. M. R. M. M.E. Cell. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). the identified caspase-3, the to elegans is the in apoptosis J. S. Miller D.K. Thornberry N.A. Yamin D.W. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, J. S. S. T. Y. Res. 1996; Google Scholar). not the nuclear protein PARP (10Nicholson D.W. All A. Thornberry N.A. Vaillancourt J.P. Ding C.K. Gallant M. Gareau Y. Griffin P.R. Labelle M. Lazebnik Y.A. Munday N.A. Raju S.M. Smulson M.E. Yamin T. Yu V.L. Miller D.K. Nature. 1995; 376: 37-43Crossref PubMed Scopus (3834) Google Scholar, M. S. Poirier G.G. Cell. 1995; Full Text PDF PubMed Scopus Google Scholar), it is capable of downstream activation of both caspase-6 S.M. M. T. G. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: PubMed Scopus (484) Google Scholar) and caspase-7 T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar) caspase-7 and caspase-6 can PARP and for between sphingolipid and the caspase cascade were investigated in this is not of that ceramide plays an role in apoptosis induced by withdrawal, or ionizing radiation, and not as a of activation of the death (reviewed in Ref. 3Hannun Y. Science. 1996; 274: 1855-1859Crossref PubMed Scopus (1510) Google Scholar). of the a or with SPP, both of which prevent apoptosis, did not with ceramide induced by (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar, D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar, J. N. L.M. Hannun Y.A. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: PubMed Scopus Google Scholar), did apoptosis induced by ceramide (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar, D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar, D.K. J. Poirier G.G. Hannun Y.A. L.M. Biochem. J. 1996; PubMed Scopus Google Scholar, J. N. L.M. Hannun Y.A. Proc. Natl. Acad. Sci. U. S. A. 1996; 93: PubMed Scopus Google Scholar, S. T. J. Kolesnick R.N. Cell 1995; Google Scholar). ceramide and apoptosis induced by Fas or can be blocked by of the D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar), in is likely caspase-8 S. M. J. Biol. Chem. 1997; PubMed Scopus Google Scholar). In we that ceramide not induce activation of caspase-8 the cell-permeable or induce activation of caspase-3 N. R. Y. S. N. 1996; PubMed Scopus Google Scholar, J. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar) and subsequent PARP D.K. J. Poirier G.G. Hannun Y.A. L.M. Biochem. J. 1996; PubMed Scopus Google Scholar) Furthermore, that exogenous C2-ceramide PARP caspase-7 the to caspase-3, through caspase-3 T. Takahashi A. R. J. Yu C.M. G. Earnshaw W.C. G. Res. 1995; Google Scholar) or we for the that C2-ceramide induces lamin B likely by caspase-6 activation The by which ceramide stimulates of caspase-6 is but it is to that this may be via caspase-3 as by and S.M. T. J. N. G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). the between ceramide and the caspase and in with other studies D.K. R. Poirier G.G. L.M. Hannun Y.A. J. 1997; PubMed Scopus Google Scholar), that ceramide can be downstream of the caspase-8 and upstream of the executioner SPP to from ceramide-mediated cell death (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar), it not it to the caspase results that of the death PARP and induced by Fas or exogenous ceramide, is by with In we that activation of the for the of these substrates by Fas ligation or ceramide is by results that SPP upstream of the by which ceramide and CED-3 further is to the for ceramide and have that of the effects of SPP are through cell (3Hannun Y. Science. 1996; 274: 1855-1859Crossref PubMed Scopus (1510) Google Scholar, 4Spiegel S. Foster D. Kolesnick R. Curr. Opin. Cell Biol. 1996; 8: 159-167Crossref PubMed Scopus (475) Google Scholar). the effects of SPP to be via intracellular as inhibition of sphingosine kinase not of SPP induced by TPA, it induces apoptosis and prevents the activity of TPA which can be by addition of SPP (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). Recently, it has been suggested that the stress-activated protein kinase pathway is required for ceramide-mediated apoptosis of of the pathway ceramide-mediated apoptosis M. R. B. W.D. Grant S. A. Kolesnick R.N. Nature. 1996; PubMed Scopus Google Scholar). has been well that Fas ligation induces activation in Jurkat T D. T. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, P. J. J. Cell. Biol. 1997; PubMed Scopus Google Scholar, 1996; PubMed Google Scholar). we have that SPP not stimulates the and it prevents activation by ceramide and apoptosis (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). Thus, we have that the of sphingolipid and of family of the protein kinase family is an factor the fate of (7Cuvillier O. Pirianov G. Kleuser B. Vanek P.J. Coso O.A. Gutkind J.S. Spiegel S. Nature. 1996; 381: 800-803Crossref PubMed Scopus (1376) Google Scholar). the between activation of the caspase cascade and the protein kinase pathway leading to apoptosis have not been a pathway leading to apoptosis, in which the and the pathway and the caspase cascade R. D. Cell. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). is that the sphingolipid ceramide and SPP, have effects on these two apoptotic downstream of of the intracellular of these to the cell for and for of
Cuvillier et al. (Thu,) studied this question.