Mitogen-activated protein kinase-mediated Fas apoptotic signaling pathway

E Goillot

J Raingeaud

A Ranger

R Tepper

R Davis

E Harlow

I Sanchez

^{Massachusetts General Hospital Cancer Center, Charlestown, MA 02129;}^{Howard Hughes Medical Institute, University of Massachusetts Medical Center, Worcester, MA 01605;}^{Department of Neurology, Harvard Medical School, Boston, MA 02115;}^{Millenium Pharmaceuticals, Inc., Cambridge, MA 02139; and}^{Department of Molecular and Cellular Biology/Biological Laboratories, Harvard University, Cambridge, MA 02138}

^{Present address: Laboratoire de Biologie Moleculaire et Cellulaire, Unité Mixte de Recherche 49, Institut National de la Recherche Agronomique LA 913, Ecole Normale Supérieure Lyon, 46, Allee d’Italie, 69364 Lyon Cedex 07, France.}

^{To whom reprint requests should be addressed at: Massachusetts General Hospital Cancer Center, Mailcode 149-7330, Building 149, 13th Street, Charlestown, MA 02129. e-mail:}ude.dravrah.hgm.xileh@wolrah.

Ed Harlow

Accepted 1996 Dec 18.

Abstract

Ligation of the cell surface receptor Fas/APO-1 (CD95) by its specific ligand or by anti-Fas antibodies rapidly induces apoptosis in susceptible cells. To characterize the molecular events involved in Fas-induced apoptosis, we examined the contribution of two subgroups of the mitogen-activated protein (MAP) kinase family, the Jun kinases or stress-activated protein kinases (JNKs/SAPKs) and the extracellular signal-regulated kinases (ERKs), in a Fas-sensitive neuroblastoma cell line. Here we show that both JNK and ERK protein kinases were activated upon Fas crosslinking through a Ras-dependent mechanism. Interference with either the JNK or ERK pathway by ectopic expression of dominant-interfering mutant proteins blocked Fas-mediated apoptosis. ERK activation was transient and associated with induced expression of the Fas receptor. In contrast, JNK activation was sustained and correlated with the onset of apoptosis. These data indicate that the ERK and the JNK groups of MAP kinases cooperate in the induction of cell death by Fas. Inhibition of Fas killing by an interleukin 1β-converting enzyme (ICE)-like protease inhibitor peptide did not modify Fas-induced JNK activation upon Fas ligation. In contrast, changes in Bcl-2 level due to expression of sense and antisense vectors influenced the sensitivity to Fas killing and Fas-induced JNK activation.

Abstract

The Fas/APO-1 protein is a 45-kDa glycosylated transmembrane protein belonging to the tumor necrosis factor (TNF) receptor family (1, 2). The Fas ligand (FasL) is a 40-kDa protein displaying significant homology to the members of the TNF family and is primarily expressed on activated T cells (3, 4). Activation of Fas by its ligand or by crosslinking with anti-Fas antibodies induces apoptosis (2, 3, 5, 6). Apoptosis induced by Fas has been shown to be involved in the regulation of lymphocyte death in the peripheral immune system (7–11) and T cell-mediated cytotoxicity (12–16).

Several gene products involved in Fas-mediated apoptosis have been characterized. The interleukin 1β-converting enzyme (ICE)-like family of cysteine proteases has been implicated in Fas killing, since ICE inhibitors and transient expression of the pox-virus-derived serpin inhibitor CrmA or an antisense ICE construct block Fas-induced apoptosis (17–19). Another enzyme implicated in Fas-induced cell death is sphingomyelinase, which hydrolyzes sphingomyelin to generate second-messenger ceramides (20–22).

Recently, Jun kinases (JNKs) have been implicated in two different models of cell death, apoptosis induced by nerve growth factor (NGF) deprivation in differentiated rat PC12 pheochromocytoma cells (23) and by environmental stresses (24, 25). JNKs can be activated by subjecting cells to environmental stresses or by exposure to pro-inflammatory cytokines, such as TNF and interleukin 1 (26–30).

The Bcl-2 oncogene, originally identified in a subset of B cell lymphomas (31–33), has been associated with resistance to apoptosis in a variety of mammalian systems, including growth factor withdrawal from hematopoietic and neuronal cells, treatment with glucocorticoids, calcium ionophores or cytotoxic drugs, and γ irradiation (34). Bcl-2 is a member of a multigene family that includes Bcl-x (35), Mcl-1 (36), A1 (37), Bax (38), Bad (39), Bak (40) in mammals, and ced-9 in the nematode Caenorhabditis elegans (41). Similarly to Bcl-2, Bcl-x_L promotes cell survival. In contrast, Bax, Bad, Bak, and Bcl-x_S, interact with Bcl-2 and/or Bcl-x_L and antagonize their protective effect. The mode of action of Bcl-2 in the inhibition of cell death is unknown.

Similar to Fas, TNFα induces cell death through interaction with its receptor. Several potent activators of JNKs, such as TNFα, cycloheximide, or exposure to heat shock, can also potentiate Fas-mediated apoptosis (5), suggesting that activation of these kinases may activate or contribute to the cell death pathway triggered by Fas. We investigated the contribution of the mitogen-activated protein (MAP) kinases to Fas-mediated killing in a Fas-sensitive neuroblastoma cell line. We found that Fas crosslinking with an anti-Fas antibody activated JNK and extracellular signal-regulated kinase (ERK) MAP kinases. Interference with either the JNK or the ERK pathway by ectopic expression of dominant-interfering proteins blocked Fas-mediated apoptosis. Inhibition of Fas killing by an ICE inhibitor did not modify Fas-induced JNK activation. In contrast, changes in Bcl-2 expression level influenced specifically Fas-stimulated JNK protein activity.

Acknowledgments

We thank G. M. Cooper for the pMMTV Ha-Ras(Asn-17) expression plasmid, R. L. Erickson for the MEK1 K→R cDNA and the anti-MEK1 antibody, A. Fattaey for the tetracycline-regulatable vector, N. G. Ahn for the ΔN3MEK expression plasmid, S. Korsmeyer for the Bcl-2 cDNA, and P. Alonso for his help with the flow cytometry analysis. L. Yamasaki, J. LaBaer, J. Koh, and E. A. Harrington are acknowledged for their careful reading of the manuscript. I.S. thanks B. Dynlacht and L. Berg for interesting discussion and for their support through the course of this endeavor. E.H. is an American Cancer Society Research Professor. R.J.D. is an investigator of the Howard Hughes Medical Institute. This work was supported by grants from the National Institutes of Health to R.I.T., E.H., and R.D.

Acknowledgments

ABBREVIATIONS

MAP	mitogen-activated protein
JNK	Jun kinase
ERK	extracellular signal-regulated kinase
ICE	interleukin 1β-converting enzyme
TNF	tumor necrosis factor
NGF	nerve growth factor
CMV	cytomegalovirus
Z-VAD-fmk	N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone

ABBREVIATIONS

References

1. Oehm A, Behrmann I, Falk W, Pawlita M, Maier G, et al J Biol Chem. 1992;267:10709–10715.[PubMed][Google Scholar]
2. Itoh N, Yonehara S, Ishii A, Yonehara M, Mizushima S, Sameshima M, Hase A, Seto Y, Nagata S. Cell. 1991;66:233–243.[PubMed]
3. Suda T, Takahashi T, Golstein P, Nagata S. Cell. 1993;75:1169–1178.[PubMed]
4. Suda T, Nagata S. J Exp Med. 1994;179:873–879.
5. Yonehara S, Ishii A, Yonehara M. J Exp Med. 1989;169:1747–1756.
6. Trauth B C, Klas C, Peters A M, Matzku S, Moller P, Falk W, Debatin K M, Krammer P H. Science. 1989;245:301–305.[PubMed]
7. Dhein J, Walczak H, Baumler C, Debatin K M, Krammer P H. Nature (London) 1995;373:438–441.[PubMed]
8. Ju S T, Panka D J, Cui H, Ettinger R, el-Khatib M, Sherr D H, Stanger B Z, Marshak-Rothstein A. Nature (London) 1995;373:444–448.[PubMed]
9. Rathmell J C, Cooke M P, Ho W Y, Grein J, Townsend S E, Davis M M, Goodnow C C. Nature (London) 1995;376:181–184.[PubMed]
10. Brunner T, Mogil R J, LaFace D, Yoo N J, Mahboubi A, Echeverri F, Martin S J, Force W R, Lynch D H, Ware C F, et al Nature (London) 1995;373:441–444.[PubMed][Google Scholar]
11. Rothstein T L, Wang J K, Panka D J, Foote L C, Wang Z, Stanger B, Cui H, Ju S T, Marshak-Rothstein A. Nature (London) 1995;374:163–165.[PubMed]
12. Walsh C M, Glass A A, Chiu V, Clark W R. J Immunol. 1994;153:2506–2514.[PubMed]
13. Stalder T, Hahn S, Erb P. J Immunol. 1994;152:1127–1133.[PubMed]
14. Rouvier E, Luciani M F, Golstein P. J Exp Med. 1993;177:195–200.
15. Lowin B, Hahne M, Mattmann C, Tschopp J. Nature (London) 1994;370:650–652.[PubMed]
16. Kagi D, Vignaux F, Ledermann B, Burki K, Depraetere V, Nagata S, Hengartner H, Golstein P. Science. 1994;265:528–530.[PubMed]
17. Los M, Van de Craen M, Penning L C, Schenk H, Westendorp M, Baeuerle P A, Droge W, Krammer P H, Fiers W, Schulze-Osthoff K. Nature (London) 1995;375:81–83.[PubMed]
18. Enari M, Hug H, Nagata S. Nature (London) 1995;375:78–81.[PubMed]
19. Tewari M, Dixit V M. J Biol Chem. 1995;270:3255–3260.[PubMed]
20. Gulbins E, Bissonnette R, Mahboubi A, Martin S, Nishioka W, Brunner T, Baier G, Baier-Bitterlich G, Byrd C, Lang F, Kolesnick A, Altman A, Green D. Immunity. 1995;2:341–351.[PubMed]
21. Cifone M G, De Maria R, Roncaioli P, Rippo M R, Azuma M, Lanier L L, Santoni A, Testi R. J Exp Med. 1994;180:1547–1552.
22. Tepper C G, Jayadev S, Liu B, Bielawska A, Wolff R, Yonehara S, Hannun Y A, Seldin M F. Proc Natl Acad Sci USA. 1995;92:8443–8447.
23. Xia Z, Dickens M, Raingeaud J, Davis R J, Greenberg M E. Science. 1995;270:1326–1331.[PubMed]
24. Verheij M, Bose R, Lin X L, Yao B, Jarvis W D, Grant S, Birrer M J, Szabo E, Zon L I, Kyriakis J M, Friedman A H, Fuks Z, Kolesnick R N. Nature (London) 1996;380:75–79.[PubMed]
25. Chen Y R, Meyer C F, Tan T H. J Biol Chem. 1996;272:631–634.[PubMed]
26. Kyriakis J M, Banerjee P, Nikolakaki E, Dai T, Rubie E A, Ahmad M F, Avruch J, Woodgett J R. Nature (London) 1994;369:156–160.[PubMed]
27. Derijard B, Hibi M, Wu I H, Barrett T, Su B, Deng T, Karin M, Davis R J. Cell. 1994;76:1025–1037.[PubMed]
28. Hibi M, Lin A, Smeal T, Minden A, Karin M. Genes Dev. 1993;7:2135–2148.[PubMed]
29. Galcheva-Gargova Z, Derijard B, Wu I H, Davis R J. Science. 1994;265:806–808.[PubMed]
30. Sluss H K, Barrett T, Derijard B, Davis R J. Mol Cell Biol. 1994;14:8376–8384.
31. Bakhshi A, Jensen J P, Goldman P, Wright J J, McBride O W, Epstein A L, Korsmeyer S J. Cell. 1985;41:899–906.[PubMed]
32. Cleary M L, Smith S D, Sklar J. Cell. 1986;47:19–28.[PubMed]
33. Tsujimoto Y, Gorham J, Cossman J, Jaffe E, Croce C M. Science. 1985;229:1390–1393.[PubMed]
34. White E. Genes Dev. 1996;10:1–15.[PubMed]
35. Boise L H, Gonzalez-Garcia M, Postema C E, Ding L, Lindsten T, Turka L A, Mao X, Nunez G, Thompson C B. Cell. 1993;74:597–608.[PubMed]
36. Kozopas K M, Yang T, Buchan H L, Zhou P, Craig R W. Proc Natl Acad Sci USA. 1993;90:3516–3520.
37. Lin E Y, Orlofsky A, Berger M S, Prystowsky M B. J Immunol. 1993;151:1979–1988.[PubMed]
38. Oltvai Z N, Milliman C L, Korsmeyer S J. Cell. 1993;74:609–619.[PubMed]
39. Yang E, Zha J, Jockel J, Boise L H, Thompson C B, Korsmeyer S J. Cell. 1995;80:285–291.[PubMed]
40. Chittenden T, Harrington E A, O’Connor R, Flemington C, Lutz R J, Evan G I, Guild B C. Nature (London) 1995;374:733–736.[PubMed]
41. Hengartner M O, Ellis R E, Horvitz H R. Nature (London) 1992;356:494–499.[PubMed]
42. Cai H, Szeberenyi J, Cooper G M. Mol Cell Biol. 1990;10:5314–5323.
43. Davis R J. Trends Biochem Sci. 1994;19:470–473.[PubMed]
44. Cobb M H, Robbins D J, Boulton T G. Curr Opin Cell Biol. 1991;3:1025–1032.[PubMed]
45. Blenis J. Proc Natl Acad Sci USA. 1993;90:5889–5892.
46. Davis R J. J Biol Chem. 1993;268:14553–14556.[PubMed]
47. Hunter T. Cell. 1995;80:225–236.[PubMed]
48. Marshall C J. Cell. 1995;80:179–185.[PubMed]
49. van den Heuvel S, Harlow E. Science. 1993;262:2050–2054.[PubMed]
50. Mansour S J, Matten W T, Hermann A S, Candia J M, Rong S, Fukasawa K, Vande Woude G F, Ahn N G. Science. 1994;265:966–970.[PubMed]
51. Fraser A, Evan G. Cell. 1996;85:781–784.[PubMed]
52. Harrington E A, Fanidi A, Evan G I. Curr Opin Genet Dev. 1994;4:120–129.[PubMed]
53. Smeyne R J, Vendrell M, Hayward M, Baker S J, Miao G G, Schilling K, Robertson L M, Curran T, Morgan J I. Nature (London) 1993;363:166–169.[PubMed]
54. Whitmarsh, A. J. & Davis, R. J. (1996) J. Mol. Med., in press. [[PubMed]
55. Tartaglia L, Ayres T, Wong G, Goeddel D. Cell. 1993;74:845–853.[PubMed]
56. Chinnaiyan A M, O’Rourke K, Tewari M, Dixit V M. Cell. 1995;81:505–512.[PubMed]
57. Boldin M P, Varfolomeev E E, Pancer Z, Mett I L, Camonis J H, Wallach D. J Biol Chem. 1995;270:7795–7798.[PubMed]
58. Stanger B Z, Leder P, Lee T H, Kim E, Seed B. Cell. 1995;81:513–523.[PubMed]
59. Boldin M P, Goncharov T M, Goltsev Y V, Wallach D. Cell. 1996;85:803–815.[PubMed]
60. Muzio M, Chinnaiyan A M, Kischkel F C, O’Rourke K, Shevchenko A, Ni J, Scaffidi C, Bretz J D, Zhang M, Gentz R, Mann M, Krammer P H, Peter M E, Dixit V M. Cell. 1996;85:817–827.[PubMed]
61. Fernandez-Sarabia M J, Bischoff J R. Nature (London) 1993;366:274–275.[PubMed]
62. Wang H G, Millan J A, Cox A D, Der C J, Rapp U R, Beck T, Zha H, Reed J C. J Cell Biol. 1995;129:1103–1114.
63. Owen-Schaub L B, Radinsky R, Kruzel E, Berry K, Yonehara S. Cancer Res. 1994;54:1580–1586.[PubMed]
64. Miura M, Zhu H, Rotello R, Hartwieg E A, Yuan J. Cell. 1993;75:653–660.[PubMed]
65. Ray C A, Black R A, Kronheim S R, Greenstreet T A, Sleath P R, Salvesen G S, Pickup D J. Cell. 1992;69:597–604.[PubMed]
66. Komiyama T, Ray C A, Pickup D J, Howard A D, Thornberry N A, Peterson E P, Salvesen G. J Biol Chem. 1994;269:19331–19337.[PubMed]
67. Gagliardini V, Fernandez P A, Lee R K, Drexler H C, Rotello R J, Fishman M C, Yuan J. Science. 1994;263:826–828.[PubMed]