Apoptosis

, Volume 6, Issue 4, pp 291–297 | Cite as

BAPTA blocks DNA fragmentation and chromatin condensation downstream of caspase-3 and DFF activation in HT-induced apoptosis in HL-60 cells

  • Min Zhang
  • Yanping Li
  • Hongqing Zhang
  • Shaobai Xue
Article

Abstract

DFF ((DNA Fragmentation Factor) is a heterodimer composed of 40 kDa (DFF40, CAD) and 45 kDa (DFF45, ICAD) subunits. During apoptosis, activated caspase-3 cleaves DFF45 and activates DFF40, a DNase that targets nucleosomal linker region and cleaves chromatin DNA into nucleosomal fragments. We have previously reported that HT induced apoptosis in HL-60 cells, and intracellular Ca2+ chelator BAPTA blocked apoptosis-associated DNA fragmentation induced by HT. We report here that HT also induced activation of caspase-3 and cleavage of DFF45. BAPTA prevented neither the caspase-3 activation nor the cleavage of DFF45. Mitochondrial membrane potential was disrupted in BAPTA-AM treated cells. However, BAPTA did prevent DNA fragmentation and chromatin condensation in HT-treated cells. These data suggest a novel role for intracellular calcium in regulating apoptotic nuclease that causes DNA fragmentation and chromatin condensation.

apoptosis Ca2+ caspase-3 DFF BAPTA-AM mitochondrial membrane potential 

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References

  1. 1.
    Wyllie AH. Glucocorticoid induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980; 284: 555-556.Google Scholar
  2. 2.
    Nagata S. Apoptosis by death factor. Cell 1997; 88: 355-365.Google Scholar
  3. 3.
    Adams JM, Cory S. The Bcl-2 protein family: Arbiters of cell survival. Science 1998; 281: 1322-1326.Google Scholar
  4. 4.
    Liu X, Kim CN, Yang J, Jemmerson R, Wang X. Induction of the apoptotic program in cell-free extracts: Requirement for dATP and cytochrome C. Cell 1996; 86: 147-157.Google Scholar
  5. 5.
    Zou H, Henzel WJ, Liu X, Lutschg A, Wang X. Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3. Cell 1997; 90: 405-413.Google Scholar
  6. 6.
    Liu X, Li P, Piotr W, Zou H, Luo X, Wang X. DNA fragmentation factor DFF40 induces DNA fragmentation and chromatin condensation during apoptosis. Proc Natl Acad Sci USA 1998; 95: 1-6.Google Scholar
  7. 7.
    Kaiser N, Edelman IS. Calcium dependence of glucocorticoid-induced lymphocytolysis. Proc Natl Acad Sci USA 1977; 74: 638-642.Google Scholar
  8. 8.
    Lam M, Dubyak G, Distelhorst CW. Effect of glucocorticoid treatment on intracellular calcium homeostasis in mouse lymphoma cells. Mol Endocrinol 1993; 7: 686-693.Google Scholar
  9. 9.
    Richterr C. Pro-oxidants and mitochondrial Ca2+: Their relationship to apoptosis and oncogenesis. FEBS Lett 1993; 325: 104-107.Google Scholar
  10. 10.
    Nicotera P, Orrenius S. The role of calcium in apoptosis. Cell Calcium 1998; 23(2-3): 173-180.Google Scholar
  11. 11.
    Gunter TE, Pfeiffer DR. Mechanisms by which mitochondria transport calcium. Am J Physiol 1990; 258: C755-C786.Google Scholar
  12. 12.
    Fang M, Zhang HQ, Xue SB. Differential effects of Ca2+ and Mg2+ on endonuclease activation in isolated promyelocytic HL-60 cell nuclei. Science in China 1998; 41(4): 351-359.Google Scholar
  13. 13.
    Fang M, Zhang HQ, Xue SB. Role of calcium in apoptosis of HL-60 cells induced by harringtonine. Science in China 1998; 41(6): 600-607.Google Scholar
  14. 14.
    Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S. A caspase-activated Dnase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 1998; 391(6662): 43-50.Google Scholar
  15. 15.
    Gaido ML, Cidlowski JA. Identification, purification, and characterization of a calcium-dependent endonuclease (NUC 18) from apoptotic rat thymocytes. J Biol Chem 1991; 266: 18580-18585.Google Scholar
  16. 16.
    Nichonova LV, Beletsky IP, Umansky SR. Properties of some nuclear nucleases of rat thymocytes and their changes in radiation-induced apoptosis. Eur J Biochem 1993; 215: 893-901.Google Scholar
  17. 17.
    Peitsch MC, Polar B, Stephan H, et al. Characterization of the endogenous deoxyyribonuclease involved in nuclear DNA degradation during apoptosis (programmed cell death). EMBO J 1993; 12: 371-377.Google Scholar
  18. 18.
    Sakahira H, Enari M, Ohsawa Y, Uchiyama Y, Nagata S. Apoptotic nuclear morphological change without DNA fragmentation. Curr Biol 1999; 20; 9(10): 543-546.Google Scholar
  19. 19.
    Liu X, Zou H, Slaughter C, Wang X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 1997; 89: 175-184.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Min Zhang
    • 1
  • Yanping Li
    • 1
  • Hongqing Zhang
    • 1
  • Shaobai Xue
    • 2
  1. 1.Department of BiologyBeijing Normal UniversityBeijingPeople's Republic of China
  2. 2.Department of BiologyBeijing Normal UniversityBeijingPeople's Republic of China

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