Abstract
Endoplasmic reticulum (ER) stress-induced apoptosis may arise from multiple environmental and pharmacological causes, but the precise mechanism(s) involved are not completely known. Members of Bcl-2 protein family are important regulators of apoptosis. In this study, we report that in a process dependent on the proapoptotic Bcl-2 members Bax and Bak, exogenously expressed fluorescent protein localized to the ER lumen is released into the cytosol in cells undergoing ER stress. Upon ER stress induction, endogenous ER luminal proteins are also released into the cytosol in a similar manner accompanied by translocation and anchorage of Bax to the ER membrane. In addition, Bax and truncated-Bid (tBid) mediate a global increase in ER membrane permeability to ER luminal proteins in vitro. Importantly, antiapoptotic Bcl-XL antagonizes the effects of proapoptotic Bcl-2 proteins on ER membrane permeability. Consistent with Bax translocation to the ER membrane in whole apoptotic cells, there is also increased tight association of Bax with the ER membrane correlated with the increase in ER membrane permeability in vitro. Overall, these data suggest that the regulation of ER membrane permeability by Bcl-2 proteins could be an important molecular mechanism of ER stress-induced apoptosis.
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Abbreviations
- ER:
-
endoplasmic reticulum
- MOM:
-
mitochondrial outer membrane
- MEF:
-
mouse embryonic fibroblast
- PDI:
-
protein disulfide isomerase
- GRP78/BiP:
-
78-kDa glucose-regulated protein
- tBid:
-
truncated-Bid
- TMRE:
-
tetramethylrhodamine, ethyl ester
References
Cory S, Adams JM . The Bcl2 family: regulators of the cellular life-or-death switch. Nat Rev Cancer 2002; 2: 647–656.
Danial NN, Korsmeyer SJ . Cell death: critical control points. Cell 2004; 116: 205–219.
Youle RJ, Strasser A . The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 2008; 9: 47–59.
Chipuk JE, Green DR . How do BCL-2 proteins induce mitochondrial outer membrane permeabilization? Trends Cell Biol 2008; 18: 157–164.
Lindsten T, Ross AJ, King A, Zong WX, Rathmell JC, Shiels HA et al. The combined functions of proapoptotic Bcl-2 family members bak and bax are essential for normal development of multiple tissues. Mol Cell 2000; 6: 1389–1399.
Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001; 292: 727–730.
Germain M, Shore GC . Cellular distribution of Bcl-2 family proteins. Sci STKE 2003; 2003: e10.
Jiang X, Wang X . Cytochrome C-mediated apoptosis. Annu Rev Biochem 2004; 73: 87–106.
Boyce M, Degterev A, Yuan J . Caspases: an ancient cellular sword of Damocles. Cell Death Differ 2004; 11: 29–37.
Oakes SA, Lin SS, Bassik MC . The control of endoplasmic reticulum-initiated apoptosis by the BCL-2 family of proteins. Curr Mol Med 2006; 6: 99–109.
Pinton P, Rizzuto R . Bcl-2 and Ca2+ homeostasis in the endoplasmic reticulum. Cell Death Differ 2006; 13: 1409–1418.
Nutt LK, Chandra J, Pataer A, Fang B, Roth JA, Swisher SG et al. Bax-mediated Ca2+ mobilization promotes cytochrome c release during apoptosis. J Biol Chem 2002; 277: 20301–20308.
Zong WX, Li C, Hatzivassiliou G, Lindsten T, Yu QC, Yuan J et al. Bax and Bak can localize to the endoplasmic reticulum to initiate apoptosis. J Cell Biol 2003; 162: 59–69.
Mathai JP, Germain M, Shore GC . BH3-only BIK regulates BAX,BAK-dependent release of Ca2+ from endoplasmic reticulum stores and mitochondrial apoptosis during stress-induced cell death. J Biol Chem 2005; 280: 23829–23836.
Rong Y, Distelhorst CW . Bcl-2 protein family members: versatile regulators of calcium signaling in cell survival and apoptosis. Annu Rev Physiol 2008; 70: 73–91.
de Brito OM, Scorrano L . Mitofusin 2 tethers endoplasmic reticulum to mitochondria. Nature 2008; 456: 605–610.
Goldstein JC, Munoz-Pinedo C, Ricci JE, Adams SR, Kelekar A, Schuler M et al. Cytochrome c is released in a single step during apoptosis. Cell Death Differ 2005; 12: 453–462.
Munoz-Pinedo C, Guio-Carrion A, Goldstein JC, Fitzgerald P, Newmeyer DD, Green DR . Different mitochondrial intermembrane space proteins are released during apoptosis in a manner that is coordinately initiated but can vary in duration. Proc Natl Acad Sci USA 2006; 103: 11573–11578.
Galluzzi L, Zamzami N, de La Motte RT, Lemaire C, Brenner C, Kroemer G . Methods for the assessment of mitochondrial membrane permeabilization in apoptosis. Apoptosis 2007; 12: 803–813.
Leber B, Lin J, Andrews DW . Embedded together: the life and death consequences of interaction of the Bcl-2 family with membranes. Apoptosis 2007; 12: 897–911.
Scorrano L, Oakes SA, Opferman JT, Cheng EH, Sorcinelli MD, Pozzan T et al. BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 2003; 300: 135–139.
Fujiki Y, Hubbard AL, Fowler S, Lazarow PB . Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum 1. J Cell Biol 1982; 93: 97–102.
Gajkowska B, Wojewodzka U, Gajda J . Translocation of Bax and Bid to mitochondria, endoplasmic reticulum and nuclear envelope: possible control points in apoptosis 1. J Mol Histol 2004; 35: 11–19.
Upton JP, Austgen K, Nishino M, Coakley KM, Hagen A, Han D et al. Caspase-2 cleavage of BID is a critical apoptotic signal downstream of endoplasmic reticulum stress. Mol Cell Biol 2008; 28: 3943–3951.
Kuwana T, Mackey MR, Perkins G, Ellisman MH, Latterich M, Schneiter R et al. Bid, Bax, and lipids cooperate to form supramolecular openings in the outer mitochondrial membrane. Cell 2002; 111: 331–342.
Smith MI, Deshmukh M . Endoplasmic reticulum stress-induced apoptosis requires bax for commitment and Apaf-1 for execution in primary neurons. Cell Death Differ 2007; 14: 1011–1019.
Li J, Lee B, Lee AS . Endoplasmic reticulum stress-induced apoptosis: multiple pathways and activation of p53-up-regulated modulator of apoptosis (PUMA) and NOXA by p53. J Biol Chem 2006; 281: 7260–7270.
Puthalakath H, O′Reilly LA, Gunn P, Lee L, Kelly PN, Huntington ND et al. ER stress triggers apoptosis by activating BH3-only protein Bim. Cell 2007; 129: 1337–1349.
Chen L, Willis SN, Wei A, Smith BJ, Fletcher JI, Hinds MG et al. Differential targeting of prosurvival Bcl-2 proteins by their BH3-only ligands allows complementary apoptotic function. Mol Cell 2005; 17: 393–403.
Kuwana T, Bouchier-Hayes L, Chipuk JE, Bonzon C, Sullivan BA, Green DR et al. BH3 domains of BH3-only proteins differentially regulate Bax-mediated mitochondrial membrane permeabilization both directly and indirectly. Mol Cell 2005; 17: 525–535.
Booth C, Koch GL . Perturbation of cellular calcium induces secretion of luminal ER proteins. Cell 1989; 59: 729–737.
Yuan XM, Li W, Dalen H, Lotem J, Kama R, Sachs L et al. Lysosomal destabilization in p53-induced apoptosis. Proc Natl Acad Sci USA 2002; 99: 6286–6291.
Ron D, Walter P . Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 2007; 8: 519–529.
Gardai SJ, McPhillips KA, Frasch SC, Janssen WJ, Starefeldt A, Murphy-Ullrich JE et al. Cell-surface calreticulin initiates clearance of viable or apoptotic cells through trans-activation of LRP on the phagocyte. Cell 2005; 123: 321–334.
Obeid M, Panaretakis T, Joza N, Tufi R, Tesniere A, van EP et al. Calreticulin exposure is required for the immunogenicity of gamma-irradiation and UVC light-induced apoptosis. Cell Death Differ 2007; 14: 1848–1850.
Burikhanov R, Zhao Y, Goswami A, Qiu S, Schwarze SR, Rangnekar VM . The tumor suppressor Par-4 activates an extrinsic pathway for apoptosis. Cell 2009; 138: 377–388.
Lum JJ, Bauer DE, Kong M, Harris MH, Li C, Lindsten T et al. Growth factor regulation of autophagy and cell survival in the absence of apoptosis. Cell 2005; 120: 237–248.
White C, Li C, Yang J, Petrenko NB, Madesh M, Thompson CB et al. The endoplasmic reticulum gateway to apoptosis by Bcl-X(L) modulation of the InsP3R. Nat Cell Biol 2005; 7: 1021–1028.
Li C, Wang X, Vais H, Thompson CB, Foskett JK, White C . Apoptosis regulation by Bcl-x(L) modulation of mammalian inositol 1,4,5-trisphosphate receptor channel isoform gating. Proc Natl Acad Sci USA 2007; 104: 12565–12570.
Bouchier-Hayes L, Munoz-Pinedo C, Connell S, Green DR . Measuring apoptosis at the single cell level. Methods 2008; 44: 222–228.
Acknowledgements
We are grateful to Drs John Eaton and Brian Wattenberg for advice. We also thank the Calcium Imaging Research Support Laboratory, Department of Physiology & Biophysics, Rosalind Franklin University. This work was supported by Schweppe Foundation and Rosalind Franklin University (CW) and NIH grants CA106599 and RR018733 and funding from JG Brown Cancer Center (CL).
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Wang, X., Olberding, K., White, C. et al. Bcl-2 proteins regulate ER membrane permeability to luminal proteins during ER stress-induced apoptosis. Cell Death Differ 18, 38–47 (2011). https://doi.org/10.1038/cdd.2010.68
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DOI: https://doi.org/10.1038/cdd.2010.68
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