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. 1996 Sep 15;318(Pt 3):749–752. doi: 10.1042/bj3180749

Chromatin condensation during apoptosis requires ATP.

G E Kass 1, J E Eriksson 1, M Weis 1, S Orrenius 1, S C Chow 1
PMCID: PMC1217681  PMID: 8836114

Abstract

The processes leading to morphological changes of the chromatin in cells that undergo apoptosis are presently unclear. We have recently shown that chromatin fragmentation and the nuclear morphological changes typically seen in apoptosis were reproduced in an in vitro system comprised of isolated rat thymocyte nuclei incubated in the presence of a lysate from Fas/APO-1-stimulated JURKAT cells [Chow, Weis, Kass, Holmström, Eriksson and Orrenius (1995) FEBS Lett. 364, 134-138]. Using this in vitro system, we now report that the presence of ATP is necessary for chromatin condensation, its movement to the nuclear periphery and apoptotic body formation. In clear contrast, chromatin cleavage into high-molecular-mass and oligonucleosomal-length DNA fragments induced by lysates derived from Fas/APO-1-activated JURKAT cells did not require the presence of ATP. The induction of these morphological changes by ATP could not be substituted by the analogues, adenosine 5'-[beta, gamma-methylene]triphosphate and adenosine 5'-[alpha, beta-methylene]-triphosphate, AMP, cAMP and UTP. However, adenosine 5'-[gamma-thio]triphosphate, and to a lesser degree GTP and ADP, could partially replace ATP in inducing nuclear apoptotic morphological changes. It is concluded that ATP is essential for the morphological changes occurring in nuclei during apoptosis, but not for DNA fragmentation.

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Selected References

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  1. Alnemri E. S., Litwack G. Activation of internucleosomal DNA cleavage in human CEM lymphocytes by glucocorticoid and novobiocin. Evidence for a non-Ca2(+)-requiring mechanism(s). J Biol Chem. 1990 Oct 5;265(28):17323–17333. [PubMed] [Google Scholar]
  2. Arends M. J., Wyllie A. H. Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol. 1991;32:223–254. doi: 10.1016/b978-0-12-364932-4.50010-1. [DOI] [PubMed] [Google Scholar]
  3. Brown D. G., Sun X. M., Cohen G. M. Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. J Biol Chem. 1993 Feb 15;268(5):3037–3039. [PubMed] [Google Scholar]
  4. Cain K., Inayat-Hussain S. H., Kokileva L., Cohen G. M. Multi-step DNA cleavage in rat liver nuclei is inhibited by thiol reactive agents. FEBS Lett. 1995 Jan 30;358(3):255–261. doi: 10.1016/0014-5793(94)01436-5. [DOI] [PubMed] [Google Scholar]
  5. Chow S. C., Weis M., Kass G. E., Holmström T. H., Eriksson J. E., Orrenius S. Involvement of multiple proteases during Fas-mediated apoptosis in T lymphocytes. FEBS Lett. 1995 May 8;364(2):134–138. doi: 10.1016/0014-5793(95)00370-o. [DOI] [PubMed] [Google Scholar]
  6. Cohen G. M., Sun X. M., Fearnhead H., MacFarlane M., Brown D. G., Snowden R. T., Dinsdale D. Formation of large molecular weight fragments of DNA is a key committed step of apoptosis in thymocytes. J Immunol. 1994 Jul 15;153(2):507–516. [PubMed] [Google Scholar]
  7. Cohen G. M., Sun X. M., Snowden R. T., Dinsdale D., Skilleter D. N. Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. Biochem J. 1992 Sep 1;286(Pt 2):331–334. doi: 10.1042/bj2860331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen J. J., Duke R. C. Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J Immunol. 1984 Jan;132(1):38–42. [PubMed] [Google Scholar]
  9. Dingwall C., Laskey R. The nuclear membrane. Science. 1992 Nov 6;258(5084):942–947. doi: 10.1126/science.1439805. [DOI] [PubMed] [Google Scholar]
  10. Enari M., Hase A., Nagata S. Apoptosis by a cytosolic extract from Fas-activated cells. EMBO J. 1995 Nov 1;14(21):5201–5208. doi: 10.1002/j.1460-2075.1995.tb00204.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Filipski J., Leblanc J., Youdale T., Sikorska M., Walker P. R. Periodicity of DNA folding in higher order chromatin structures. EMBO J. 1990 Apr;9(4):1319–1327. doi: 10.1002/j.1460-2075.1990.tb08241.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fry A. M., Schultz S. J., Bartek J., Nigg E. A. Substrate specificity and cell cycle regulation of the Nek2 protein kinase, a potential human homolog of the mitotic regulator NIMA of Aspergillus nidulans. J Biol Chem. 1995 May 26;270(21):12899–12905. doi: 10.1074/jbc.270.21.12899. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Lazebnik Y. A., Cole S., Cooke C. A., Nelson W. G., Earnshaw W. C. Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis. J Cell Biol. 1993 Oct;123(1):7–22. doi: 10.1083/jcb.123.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lu K. P., Hunter T. Evidence for a NIMA-like mitotic pathway in vertebrate cells. Cell. 1995 May 5;81(3):413–424. doi: 10.1016/0092-8674(95)90394-1. [DOI] [PubMed] [Google Scholar]
  16. Martin S. J., Newmeyer D. D., Mathias S., Farschon D. M., Wang H. G., Reed J. C., Kolesnick R. N., Green D. R. Cell-free reconstitution of Fas-, UV radiation- and ceramide-induced apoptosis. EMBO J. 1995 Nov 1;14(21):5191–5200. doi: 10.1002/j.1460-2075.1995.tb00203.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. O'Connell M. J., Norbury C., Nurse P. Premature chromatin condensation upon accumulation of NIMA. EMBO J. 1994 Oct 17;13(20):4926–4937. doi: 10.1002/j.1460-2075.1994.tb06820.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Oberhammer F., Wilson J. W., Dive C., Morris I. D., Hickman J. A., Wakeling A. E., Walker P. R., Sikorska M. Apoptotic death in epithelial cells: cleavage of DNA to 300 and/or 50 kb fragments prior to or in the absence of internucleosomal fragmentation. EMBO J. 1993 Sep;12(9):3679–3684. doi: 10.1002/j.1460-2075.1993.tb06042.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shi L., Nishioka W. K., Th'ng J., Bradbury E. M., Litchfield D. W., Greenberg A. H. Premature p34cdc2 activation required for apoptosis. Science. 1994 Feb 25;263(5150):1143–1145. doi: 10.1126/science.8108732. [DOI] [PubMed] [Google Scholar]
  20. Sorenson C. M., Barry M. A., Eastman A. Analysis of events associated with cell cycle arrest at G2 phase and cell death induced by cisplatin. J Natl Cancer Inst. 1990 May 2;82(9):749–755. doi: 10.1093/jnci/82.9.749. [DOI] [PubMed] [Google Scholar]
  21. Sun X. M., Cohen G. M. Mg(2+)-dependent cleavage of DNA into kilobase pair fragments is responsible for the initial degradation of DNA in apoptosis. J Biol Chem. 1994 May 27;269(21):14857–14860. [PubMed] [Google Scholar]
  22. Sun X. M., Snowden R. T., Dinsdale D., Ormerod M. G., Cohen G. M. Changes in nuclear chromatin precede internucleosomal DNA cleavage in the induction of apoptosis by etoposide. Biochem Pharmacol. 1994 Jan 20;47(2):187–195. doi: 10.1016/0006-2952(94)90005-1. [DOI] [PubMed] [Google Scholar]
  23. Trauth B. C., Klas C., Peters A. M., Matzku S., Möller P., Falk W., Debatin K. M., Krammer P. H. Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science. 1989 Jul 21;245(4915):301–305. doi: 10.1126/science.2787530. [DOI] [PubMed] [Google Scholar]
  24. Tsukiyama T., Becker P. B., Wu C. ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature. 1994 Feb 10;367(6463):525–532. doi: 10.1038/367525a0. [DOI] [PubMed] [Google Scholar]
  25. Varga-Weisz P. D., Blank T. A., Becker P. B. Energy-dependent chromatin accessibility and nucleosome mobility in a cell-free system. EMBO J. 1995 May 15;14(10):2209–2216. doi: 10.1002/j.1460-2075.1995.tb07215.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Walker P. R., Weaver V. M., Lach B., LeBlanc J., Sikorska M. Endonuclease activities associated with high molecular weight and internucleosomal DNA fragmentation in apoptosis. Exp Cell Res. 1994 Jul;213(1):100–106. doi: 10.1006/excr.1994.1178. [DOI] [PubMed] [Google Scholar]
  27. Weis M., Schlegel J., Kass G. E., Holmström T. H., Peters I., Eriksson J., Orrenius S., Chow S. C. Cellular events in Fas/APO-1-mediated apoptosis in JURKAT T lymphocytes. Exp Cell Res. 1995 Aug;219(2):699–708. doi: 10.1006/excr.1995.1281. [DOI] [PubMed] [Google Scholar]
  28. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  29. Yonehara S., Ishii A., Yonehara M. A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med. 1989 May 1;169(5):1747–1756. doi: 10.1084/jem.169.5.1747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zhivotovsky B., Cedervall B., Jiang S., Nicotera P., Orrenius S. Involvement of Ca2+ in the formation of high molecular weight DNA fragments in thymocyte apoptosis. Biochem Biophys Res Commun. 1994 Jul 15;202(1):120–127. doi: 10.1006/bbrc.1994.1901. [DOI] [PubMed] [Google Scholar]

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