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Upregulation of alpha globin promotes apoptotic cell death in the hematopoietic cell line FL5.12

Apoptosis volume 10, pages 1043–1062 (2005)Cite this article

Abstract

The function of alpha globin in the context of oxygen transport in erythroid cells is well described. Recently the expression of alpha globin was shown to be upregulated upon specific apoptotic stimuli like cytokine deprivation or cisplatin treatment in the hematopoietic pro-B cell line, FL5.12. In contrast to alpha globin, beta globin or globin-like genes were expressed at a very low level or were not expressed at all. Further, we found that alpha globin was not associated with heme. Apoptotic cells neither produced hemoglobin nor displayed a phenotype of cells differentiating down the erythroid lineage. Also other cell lines of variable differentiation status (NIH3T3, HeLa, K562) upregulated alpha globin during treatment with apoptosis-inducing agents. Under IL-3-deprived conditions GFP-alpha globin accelerated the progression of apoptosis comparable to GFP-Bax. GFP-alpha globin was expressed at a low level and enrichment of FL5.12 cells expressing GFP-alpha globin was difficult even in the presence of IL-3. Caspase-8, -9 and -3 as well as the proapoptotic factor Bax and cytochrome c were activated. Antisense alpha globin downregulated the expression of endogenous alpha globin und reduced caspase activity. Taken together these data indicate that alpha globin is a new and crucial factor in apoptosis especially supporting the mitochondrial pathway.

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References

  1. Adachi-Yamada T, O’Connor MB. Mechanisms for removal of developmentally abnormal cells: cell competition and morphogenetic apoptosis. J Biochem (Tokyo) 2004; 136(1): 13–17.

    Google Scholar 

  2. Khanna KK, Lavin MF. Ionizing radiation and UV induction of p53 protein by different pathways in ataxia-telangiectasia cells. Oncogene 1993; 8(12): 3307–3312.

    PubMed  Google Scholar 

  3. Selvakumaran M, Lin HK, Miyashita T, Wang HG, Krajewski S, Reed JC, et al. Immediate early up-regulation of bax expression by p53 but not TGF beta 1: A paradigm for distinct apoptotic pathways. Oncogene 1994; 9(6): 1791–1798.

    PubMed  Google Scholar 

  4. Boatright KM, Salvesen GS. Mechanisms of caspase activation. Curr Opin Cell Biol 2003; 15(6): 725–731.

    Article  PubMed  Google Scholar 

  5. Kuwana T, Smith JJ, Muzio M, Dixit V, Newmeyer DD, Kornbluth S. Apoptosis induction by caspase-8 is amplified through the mitochondrial release of cytochrome c. J Biol Chem 1998; 273(26): 16589–16594.

    Article  PubMed  Google Scholar 

  6. Shelton JG, Steelman LS, Lee JT, Knapp SL, Blalock WL, Moye PW, et al. Effects of the RAF/MEK/ERK and PI3K/AKT signal transduction pathways on the abrogation of cytokine-dependence and prevention of apoptosis in hematopoietic cells. Oncogene 2003; 22(16): 2478–2492.

    Article  PubMed  Google Scholar 

  7. Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E, et al. Regulation of cell death protease caspase-9 by phosphorylation. Science 1998; 282(5392): 1318–1321.

    Google Scholar 

  8. Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS et al. Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell 1999; 96(6): 857–868.

    Article  PubMed  Google Scholar 

  9. Muirhead H, Cox JM, Mazzarella L, Perutz MF. Structure and function of haemoglobin. 3. A three-dimensional fourier synthesis of human deoxyhaemoglobin at 5.5 Angstrom resolution. J Mol Biol 1967; 28(1): 117–156.

    PubMed  Google Scholar 

  10. Rabbitts TH. Bacterial cloning of plasmids carrying copies of rabbit globin messenger RNA. Nature 1976; 260(5548): 221–225.

    Article  PubMed  Google Scholar 

  11. Hardison RC. A brief history of hemoglobins: Plant, animal, protist, and bacteria. Proc Natl Acad Sci USA 1996; 93(12): 5675–5679.

    Article  PubMed  Google Scholar 

  12. Fessas P, Loukopoulos D, Kaltsoya A. Peptide analysis of the inclusions of erythroid cells in beta-thalassemia. Biochim Biophys Acta 1966; 124(2): 430–432.

    PubMed  Google Scholar 

  13. Kihm AJ, Kong Y, Hong W, Russell JE, Rouda S, Adachi K, et al. An abundant erythroid protein that stabilizes free alpha-haemoglobin. Nature 2002; 417(6890): 758–763.

    Article  PubMed  Google Scholar 

  14. Wride MA, Mansergh FC, Adams S, Everitt R, Minnema SE, Rancourt DE, et al. Expression profiling and gene discovery in the mouse lens. Mol Vis 2003; 9: 360–96:360-396.

    PubMed  Google Scholar 

  15. Brachat A, Pierrat B, Brungger A, Heim J. Comparative microarray analysis of gene expression during apoptosis-induction by growth factor deprivation or protein kinase C inhibition. Oncogene 2000; 19(44): 5073–5082.

    Article  PubMed  Google Scholar 

  16. Devireddy LR, Teodoro JG, Richard FA, Green MR. Induction of apoptosis by a secreted lipocalin that is transcriptionally regulated by IL-3 deprivation. Science 2001; 293(5531): 829–834.

    Article  PubMed  Google Scholar 

  17. Brachat A, Pierrat B, Xynos A, Brecht K, Simonen M, Brungger A et al. A microarray-based, integrated approach to identify novel regulators of cancer drug response and apoptosis. Oncogene 2002; 21(54): 8361–8371.

    Article  PubMed  Google Scholar 

  18. Taketani S, Immenschuh S, Go S, Sinclair PR, Stockert RJ, Liem HH, et al. Hemopexin from four species inhibits the association of heme with cultured hepatoma cells or primary rat hepatocytes exhibiting a small number of species specific hemopexin receptors. Hepatology 1998; 27(3): 808–814.

    Article  PubMed  Google Scholar 

  19. Mahadevappa M, Warrington JA. A high-density probe array sample preparation method using 10-to 100-fold fewer cells. Nat Biotechnol 1999; 17(11): 1134–1136.

    Article  PubMed  Google Scholar 

  20. Jung M, Sabat R, Kratzschmar J, Seidel H, Wolk K, Schonbein C, et al. Expression profiling of IL-10-regulated genes in human monocytes and peripheral blood mononuclear cells from psoriatic patients during IL-10 therapy. Eur J Immunol 2004; 34(2): 481–493.

    Article  PubMed  Google Scholar 

  21. Sassa S. Sequential induction of heme pathway enzymes during erythroid differentiation of mouse Friend leukemia virus-infected cells. J Exp Med 1976; 143(2): 305–315.

    Article  PubMed  Google Scholar 

  22. Worthington RE, Bossie-Codreanu J, Van Zant G. Quantitation of erythroid differentiation in vitro using a sensitive colorimetric assay for hemoglobin. Exp Hematol 1987; 15(1): 85–92.

    PubMed  Google Scholar 

  23. Swift, Lorens, Achacoso, Nolan. Retroviral transduction. Current Protocols in Immunology 1999, Unit 10, 28(Suppl).

  24. Coyle B, Freathy C, Gant TW, Roberts RA, Cain K. Characterization of the transforming growth factor-beta 1-induced apoptotic transcriptome in FaO hepatoma cells. J Biol Chem 2003; 278(8): 5920–5928.

    Article  PubMed  Google Scholar 

  25. Burmester T, Weich B, Reinhardt S, Hankeln T. A vertebrate globin expressed in the brain. Nature 2000; 407(6803): 520–523.

    Article  PubMed  Google Scholar 

  26. Burmester T, Ebner B, Weich B, Hankeln T. Cytoglobin: A novel globin type ubiquitously expressed in vertebrate tissues. Mol Biol Evol 2002; 19(4): 416–421.

    PubMed  Google Scholar 

  27. Rutherford T, Clegg JB, Higgs DR, Jones RW, Thompson J, Weatherall DJ. Embryonic erythroid differentiation in the human leukemic cell line K562. Proc Natl Acad Sci USA 1981; 78(1): 348–352.

    PubMed  Google Scholar 

  28. Bertrand R, Solary E, O’Connor P, Kohn KW, Pommier Y. Induction of a common pathway of apoptosis by staurosporine. Exp Cell Res 1994; 211(2): 314–321.

    Article  PubMed  Google Scholar 

  29. Abreu-Martin MT, Vidrich A, Lynch DH, Targan SR. Divergent induction of apoptosis and IL-8 secretion in HT-29 cells in response to TNF-alpha and ligation of Fas antigen.J Immunol 1995; 155(9): 4147–4154.

    PubMed  Google Scholar 

  30. Johnson BW, Boise LH. Bcl-2 and caspase inhibition cooperate to inhibit tumor necrosis factor-alpha-induced cell death in a Bcl-2 cleavage-independent fashion. J Biol Chem 1999; 274(26): 18552–18558.

    Article  PubMed  Google Scholar 

  31. Fulda S, Meyer E, Debatin KM. Metabolic inhibitors sensitize for CD95 (APO-1/Fas)-induced apoptosis by down-regulating Fas-associated death domain-like interleukin 1-converting enzyme inhibitory protein expression. Cancer Res 2000; 60(14): 3947–3956.

    PubMed  Google Scholar 

  32. Siddik ZH. Cisplatin: mode of cytotoxic action and molecular basis of resistance. Oncogene 2003; 22(47): 7265–7279.

    Article  PubMed  Google Scholar 

  33. Jiang S, Cai J, Wallace DC, Jones DP. Cytochrome c-mediated apoptosis in cells lacking mitochondrial DNA. Signaling pathway involving release and caspase 3 activation is conserved. J Biol Chem 1999; 274(42): 29905–29911.

    Article  PubMed  Google Scholar 

  34. Shibahara S, Muller RM, Taguchi H. Transcriptional control of rat heme oxygenase by heat shock. J Biol Chem 1987; 262(27): 12889–12892.

    Google Scholar 

  35. Aries A, Trentesaux C, Ottolenghi S, Jardillier JC, Jeannesson P, Doubeikovski A. Activation of erythroid-specific promoters during anthracycline-induced differentiation of K562 cells. Blood 1996; 87(7): 2885–2890.

    PubMed  Google Scholar 

  36. Mc Gee MM, Campiani G, Ramunno A, Fattorusso C, Nacci V, Lawler M et al. Pyrrolo-1,5-benzoxazepines induce apoptosis in chronic myelogenous leukemia (CML) cells by bypassing the apoptotic suppressor bcr-abl. J Pharmacol Exp Ther 2001; 296(1): 31–40.

    PubMed  Google Scholar 

  37. Dedoussis GV, Mouzaki A, Theodoropoulou M, Menounos P, Kyrtsonis MC, Karameris A et al. Endogenous interleukin 6 conveys resistance to cis-diamminedichloroplatinum-mediated apoptosis of the K562 human leukemic cell line. Exp Cell Res 1999; 249(2): 269–278.

    Article  Google Scholar 

  38. Luo X, Budihardjo I, Zou H, Slaughter C, Wang X. Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 1998; 94(4): 481–490.

    PubMed  Google Scholar 

  39. McKearn JP, McCubrey J, Fagg B. Enrichment of hematopoietic precursor cells and cloning of multipotential B-lymphocyte precursors. Proc Natl Acad Sci USA 1985; 82(21): 7414–7418.

    PubMed  Google Scholar 

  40. Ahmed MM. Regulation of radiation-induced apoptosis by early growth response-1 gene in solid tumors. Curr Cancer Drug Targets 2004; 4(1): 43–52.

    Article  PubMed  Google Scholar 

  41. Morceau F, Chenais B, Gillet R, Jardillier J C, Jeannesson P, Trentesaux C. Transcriptional and posttranscriptional regulation of erythroid gene expression in anthracycline-induced differentiation of human erythroleukemic cells. Cell Growth Differ 1996; 7(8): 1023–1029.

    PubMed  Google Scholar 

  42. Scholander Pf. Oxygen transport through hemoglobin solutions. Science 1960; 131: 585–90: 585–590.

    PubMed  Google Scholar 

  43. Yoshida E, Ohmura S, Sugiki M, Anai K, Maruyama M. A novel function of extraerythrocytic hemoglobin: Identification of globin as a stimulant of plasminogen activator biosynthesis in human fibroblasts. Thromb Haemost 2001; 86(6): 1521–1527.

    PubMed  Google Scholar 

  44. Spadaccini A, Tilbrook PA, Sarna MK, Crossley M, Bieker JJ, Klinken SP. Transcription factor erythroid Kruppel-like factor (EKLF) is essential for the erythropoietin-induced hemoglobin production but not for proliferation, viability, or morphological maturation. J Biol Chem 1998; 273(37): 23793–23798.

    Article  PubMed  Google Scholar 

  45. Yuan J, Bunyaratvej A, Fucharoen S, Fung C, Shinar E, Schrier SL. The instability of the membrane skeleton in thalassemic red blood cells. Blood 1995; 86(10): 3945–3950.

    PubMed  Google Scholar 

  46. Cardone MH, Roy N, Stennicke HR, Salvesen GS, Franke TF, Stanbridge E et al. Regulation of cell death protease caspase-9 by phosphorylation. Science 1998; 282(5392): 1318–1321.

    Google Scholar 

  47. Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, et al. Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 1997; 91(2): 231–241.

    Article  PubMed  Google Scholar 

  48. Bojes HK, Feng X, Kehrer JP, Cohen GM. Apoptosis in hematopoietic cells (FL5.12) caused by interleukin-3 withdrawal: Relationship to caspase activity and the loss of glutathione. Cell Death Differ 1999; 6(1): 61–70.

    Article  PubMed  Google Scholar 

  49. Zou H, Li Y, Liu X, Wang X. An APAF-1.cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J Biol Chem 1999; 274(17): 11549–11556.

    Google Scholar 

  50. Skurk C, Maatz H, Kim HS, Yang J, Abid MR, Aird WC et al. The Akt-regulated forkhead transcription factor FOXO3a controls endothelial cell viability through modulation of the caspase-8 inhibitor FLIP. J Biol Chem 2004; 279(2): 1513–1525.

    Article  PubMed  Google Scholar 

  51. Johnson DE. Regulation of survival pathways by IL-3 and induction of apoptosis following IL-3 withdrawal. Front Biosci 1998; 3: d313–24: d313–d324.

    PubMed  Google Scholar 

  52. Kumar S. Inhibition of apoptosis by the expression of antisense Nedd2. FEBS Lett 1995; 368(1): 69–72.

    Article  PubMed  Google Scholar 

  53. Lassus P, Opitz-Araya X, Lazebnik Y. Requirement for caspase-2 in stress-induced apoptosis before mitochondrial permeabilization. Science 2002; 297(5585): 1352–1354.

    Article  PubMed  Google Scholar 

  54. Troy CM, Shelanski ML. Caspase-2 redux. Cell Death Differ 2003; 10(1): 101–107.

    Article  PubMed  Google Scholar 

  55. Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD et al. Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, $-$7, $-$8, and $-$10 in a caspase-9-dependent manner. J Cell Biol 1999; 144(2): 281–292.

    Article  PubMed  Google Scholar 

  56. Porter AG, Janicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death Differ 1999; 6(2): 99–104.

    Article  PubMed  Google Scholar 

  57. Eskes R, Desagher S, Antonsson B, Martinou JC. Bid induces the oligomerization and insertion of Bax into the outer mitochondrial membrane. Mol Cell Biol 2000; 20(3): 929–935.

    Article  PubMed  Google Scholar 

  58. Finucane DM, Bossy-Wetzel E, Waterhouse NJ, Cotter TG, Green DR. Bax-induced caspase activation and apoptosis via cytochrome c release from mitochondria is inhibitable by Bcl-xL. J Biol Chem 1999; 274(4): 2225–2233.

    Article  PubMed  Google Scholar 

  59. Zha J, Harada H, Yang E, Jockel J, Korsmeyer SJ. Serine phosphorylation of death agonist BAD in response to survival factor results in binding to 14-3-3 not BCL-X(L). Cell 1996; 87(4): 619–628.

    Article  PubMed  Google Scholar 

  60. Murphy KM, Ranganathan V, Farnsworth ML, Kavallaris M, Lock RB. Bcl-2 inhibits Bax translocation from cytosol to mitochondria during drug-induced apoptosis of human tumor cells. Cell Death Differ 2000; 7(1): 102–111.

    Article  PubMed  Google Scholar 

  61. Liu X, Kim CN, Yang J, Jemmerson R, Wang X. Induction of apoptotic program in cell-free extracts: Requirement for dATP and cytochrome c. Cell 1996; 86(1): 147–157.

    Article  PubMed  Google Scholar 

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Author notes

  1. J. Heim

    Present address: Basilea Pharmaceutica AG, Grenzacherstrasse 487, CH-4005, Basel, Switzerland

Affiliations

  1. Novartis Institutes for BioMedical Research Basel, CH-4002, Basel, Switzerland

    K. Brecht, M. Simonen & J. Heim

  2. Zentrallabor Stadtspital Triemli Zürich, Switzerland

    K. Brecht

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Brecht, K., Simonen, M. & Heim, J. Upregulation of alpha globin promotes apoptotic cell death in the hematopoietic cell line FL5.12. Apoptosis 10, 1043–1062 (2005). https://doi.org/10.1007/s10495-005-0617-9

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Keywords

  • alpha globin
  • apoptosis
  • caspases
  • heme