Abstract
Apoptosis is regulated by the action of the Bcl-2 family of proteins, which includes anti- and pro-apoptotic members such as Bcl-xS and Bax. These proteins may differ from each other in structure, mechanism of action and interactions with anti-apoptotic signaling. The mechanism whereby Bax induces cell death has been studied in some cellular systems, but the mechanism of Bcl-xS-induced apoptosis is largely unknown. In this study we investigated and compared the apoptotic effects of Bcl-xS and Bax in the pheochromocytoma cell line, PC12 (a useful model system for studying neuronal apoptosis), and the extent to which they are protected by the survival factor, nerve growth factor (NGF). PC12 cells express endogenous Bcl-xS, Bax and Bcl-xL proteins. Subcellular fractionation revealed that Bax is presented mainly in the cytosolic and the heavy membrane fractions, Bcl-xS is present only in the cytosol, and the anti-apoptotic protein Bcl-xL is located mainly in the heavy membrane fraction. In contrast to the cytosolic localization of endogenous Bcl-xS, the exogenously overexpressed Bcl-xS is localized to the mitochondria. Overexpression of Bcl-xS or Bax induces cell death in the transfected cells. The cell death induced by overexpression of Bcl-xS was inhibited by co-expression of Bcl-xS with Bcl-2 or Bcl-xL, or by treatment with the broad-spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoro-methylketone (Z-VAD-FMK) or with NGF. The Bcl-2 mutants ΔC22, which lacks the transmembrane domain, and G145A (mI-3) were able to inhibit the death-inducing effect of Bcl-xS. These results therefore suggest that the apoptotic pathway induced by overexpression of Bcl-xS in PC12 cells can be controlled by Bcl-2 and Bcl-xL, is mediated by caspases, and can be inhibited by the NGF signaling pathway. The Bax-induced cell death was inhibited by co-expression of Bax with Bcl-2 or Bcl-xL, but was not inhibited by Z-VAD-FMK, NGF, or the Bcl-2 mI-3 or ΔC22 mutants. These results therefore suggest that Bax induces a caspase-independent cell death pathway which is blocked by Bcl-2 but not by the NGF signaling pathway. They further suggest that Bcl-xS and Bax induce different cell death pathways in PC12 cells.
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References
Barinaga M . 1998 Science 280: 32–34
Berger J, Hauber J, Hauber R, Geiger R and Cullen BR . 1988 Gene 66: 1–10
Boise LH, González-García M, Postema CE, Ding L, Lindsten T, Turka LA, Mao X, Nuñez G and Thompson CB . 1993 Cell 74: 597–608
Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, Frisch S and Reed JC . 1998 Science 282: 1318–1321
Chao DT and Korsmeyer SJ . 1998 Annu Rev Immunol 16: 395–419
Cheng EH-Y, Levine B, Boise LH, Thompson CB and Hardwick JM . 1996 Nature 379: 554–556
Cheng EH, Kirsch DG, Clem RJ, Ravi R, Kastan MB, Bedi A, Ueno K and Hardwick JM . 1997 Science 278: 1966–1968
Chomczynski P and Sacchi N . 1987 Anal Biochem 162: 156–159
Clarke MF, Apel IJ, Benedict MA, Eipers PG, Sumantran V, González-García M, Doedens M, Fukunaga N, Davidson B, Dick JE, Minn AJ, Boise LH, Thompson CB, Wicha M and Nuñez G . 1995 Proc Natl Acad Sci USA 92: 11024–11028
Cohen GM . 1997 Biochem J 326: 1–16
Crowder RJ and Freeman RS . 1998 J Neurosci 18: 2933–2943
Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y and Greenberg ME . 1997 Cell 91: 231–241
Dixon EP, Stephenson DT, Clemens JA and Little SP . 1997 Brain Res 776: 222–229
Dole MG, Clarke MF, Holman P, Benedict M, Lu J, Jasty R, Eipers P, Thompson CB, Rode C, Bloch C, Nuñez G and Castle VP . 1996 Cancer Res 56: 5734–5740
Ealovega MK, McGinnis PK, Sumantran VN, Clarke MF and Wicha MS . 1996 Cancer Res 56: 1965–1969
Evan G and Littlewood T . 1998 Science 281: 1317–1322
Farrow SN and Brown R . 1996 Curr Opin Genet Dev 6: 45–49
Finucane DM, Bossy-Wetzel E, Waterhouse NJ, Cotter TG and Green DR . 1999 J Biol Chem 274: 2225–2233
González-García M, Pérez-Ballestero R, Ding L, Duan L, Boise LH, Thompson CB and Nuñez G . 1994 Development 120: 3033–3042
Goping IS, Gross A, Lavoie JN, Nguyen M, Jemmerson R, Roth K, Korsmeyer SJ and Shore GC . 1998 J Cell Biol 143: 207–215
Green DR and Martin SJ . 1995 Curr Opin Immunol 7: 694–703
Gross A, Jockel J, Wei MC and Korsmeyer SJ . 1998 EMBO J 17: 3878–3885
Haviv R, Lindenboim L, Li H, Yuan J and Stein R . 1998 J Neurosci Res 52: 491–497
Heermeier K, Benedict M, Li M, Furth P, Nuñez G and Hennighausen L . 1996 Mech Dev 56: 197–207
Hegde R, Srinivasula SM, Ahmad M, Fernandes-Alnemri T and Alnemri ES . 1998 J Biol Chem 273: 7783–7786
Hemmings BA . 1997 Science 275: 665–668
Horiuchi M, Hayashida W, Kambe T, Yamada T and Dzau VJ . 1997 J Biol Chem 272: 19022–19026
Hsu SY, Kaipia A, McGee E, Lomeli M and Hsueh AJW . 1997 Proc Natl Acad Sci USA 94: 12401–12406
Igase M, Okura T, Kitami Y and Hiwada K . 1999 Clin Sci 96: 605–612
Inohara N, Ekhterae D, Garcia I, Carrio R, Merino J, Merry A, Chen S and Nuñez G . 1998 J Biol Chem 273: 8705–8710
Inohara N, Gourley TS, Carrio R, Muniz M, Merino J, Garcia I, Koseki T, Hu Y, Chen S and Nunez G . 1998 J Biol Chem 273: 32479–32486
Jacobson MD . 1997 Curr Biol 7: R277–R281
Jia L, Macey MG, Yin Y, Newland AC and Kelsey SM . 1999 Blood 93: 2353–2359
Katoh S, Mitsui Y, Kitani K and Suzuki T . 1996 Biochem Biophys Res Commun 229: 653–657
Kelekar A and Thompson CB . 1998 Trends Cell Biol 8: 324–330
Li H, Zhu H, Xu CJ and Yuan J . 1998 Cell 94: 491–501
Li H and Yuan J . 1999 Curr Opin Cell Biol 11: 261–266
Lindenboim L, Haviv R and Stein R . 1995 J Neurosci Res 64: 1054–1063
Maroto R and Perez-Polo JR . 1997 J Neurochem 69: 514–523
Martinou I, Fernandez P-A, Missotten M, White E, Allet B, Sadoul R and Martinou J-C . 1995 J Cell Biol 128: 102–208
Martinou I, Missotten M, Fernandez PA, Sadoul R and Martinou JC . 1998 Neuroreport 9: 15–19
McCarthy NJ, Whyte MK, Gilbert CS and Evan GI . 1997 J Cell Biol 136: 215–227
Middleton G, Nunez G and Davies AM . 1996 Development 122: 695–701
Minn AJ, Boise LH and Thompson CB . 1996 J Biol Chem 271: 6306–6312
Monney L, Otter I, Olivier R, Ozer HL, Haas AL, Omura S and Borner C . 1998 J Biol Chem 273: 6121–6131
Nicholson DW and Thornberry NA . 1997 Trends Biochem Sci 22: 299–306
O'Connor L, Strasser A, O'Reilly LA, Hausmann G, Adams JM, Cory S and Huang DCS . 1998 EMBO J 17: 384–395
Okuno S, Shimizu S, Ito T, Nomura M, Hamada E, Tsujimoto Y and Matsuda H . 1998 J Biol Chem 273: 34272–34277
Oltvai ZN and Korsmeyer SJ . 1994 Cell 79: 189–192
Oltvai ZN, Milliman CL and Korsmeyer SJ . 1993 Cell 74: 609–619
Orth K and Dixit VM . 1997 J Biol Chem 272: 8841–8844
Pan G, O'Rourke K and Dixit VM . 1998 J Biol Chem 273: 5841–5845
Pastorino JG, Chen ST, Tafani M, Snyder JW and Farber JL . 1998 J Biol Chem 273: 7770–7775
Pittman RN, Wang SW, Dibenedetto AJ and Mills JC . 1993 J Neurosci Res 13: 3669–3680
Reed JC . 1998 Oncogene 17: 3225–3236
Rukenstein A, Rydel RE and Greene LA . 1991 J Neurosci Res 11: 2552–2563
Salvesen GS and Dixit VM . 1997 Cell 91: 443–446
Seto M, Jaeger U, Hockett RD, Graninger W, Bennett S, Goldman P and Korsmeyer SJ . 1998 EMBO J 7: 123–131
Song Q, Kuang Y, Dixit VM and Vincenz C . 1999 EMBO J 18: 167–178
Spear N, Estevez AG, Barbeito L, Beckman JS and Johnson GV . 1997 J Neurochem 69: 53–59
St Clair EG, Anderson SJ and Oltvai ZN . 1997 J Biol Chem 272: 29347–29355
Stefanis L, Park DS, Yan CYI, Farinelli SE, Tory CM, Shelanski ML and Greene LA . 1996 J Biol Chem 271: 30663–30671
Stefanis L, Troy CM, Qi H, Shelanski ML and Greene LA . 1998 J Neurosci 18: 9204–9215
Strasser A, Huang DCS and Vaux DL . 1997 Biochim Biophys Acta 1333: F151–F178
Wang K, Yin X-M, Chao DT, Milliman CL and Korsmeyer SJ . 1996 Genes and Dev 10: 2859–2869
Xiang J, Chao DT and Korsmeyer SJ . 1996 Proc Natl Acad Sci USA 93: 14559–14563
Yao R and Cooper GM . 1995 Science 267: 2003–2006
Yin X-M, Oltvai ZN and Korsmeyer SJ . 1994 Nature 369: 321–323
Acknowledgements
We thank Dr Craig Thompson for providing the Bcl-xL and Bcl-xS vectors, Dr Stanley Korsmeyer for providing the Bax, Bcl-2 and Bcl-2 mutant vectors and Dr Ron Kopito for providing anti-calnexin antibodies. We are also grateful to Ms Shirley Smith for excellent editorial assistance. This work was supported by the United States Israel–Binational Science Foundation.
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Lindenboim, L., Yuan, J. & Stein, R. Bcl-xS and Bax induce different apoptotic pathways in PC12 cells. Oncogene 19, 1783–1793 (2000). https://doi.org/10.1038/sj.onc.1203495
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DOI: https://doi.org/10.1038/sj.onc.1203495
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