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Gamma irradiation-induced apoptosis in murine pre-B cells prevents the condensation of fibrillar chromatin in early S phase

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Abstract

Local changes in chromatin structure leading to temporally distinct geometric forms were characterized in nuclei of reversibly permeabilized cells. Reversal of permeabilization was tested by 3H-thymidine incorporation and trypan blue dye exclusion. Apoptotic changes were visualized in a cell cycle dependent manner at the chromatin level by fluorescent microscopy in non-irradiated cells and after 400 rad Co60 irradiation. Fluorescent microscopy of chromatin structures belonging mainly to the interphase of the cell cycle confirmed the existence of specific geometric forms in nuclei of non-irradiated cells. In this control population, the following main transitory forms of condensing chromatin were distinguished: decondensed veil-like structures and fibrous structures in early and mid S phase (2.0–2.5 average C-value), chromatin bodies, semicircles later in mid S phase (3.0–3.5 C), precondensed chromosomes in late S (3.5–3.7 C) and metaphase chromosomes at the end and after S phase (3.7–4.0 C). Our results show that upon γ-irradiation (a) the cellular and nuclear sizes were increased, (b) the DNA content was lower in each elutriated subpopulation of cells, (c) the progression of the cell cycle was arrested in the early S phase at 2.4 C value, (d) the chromatin condensation was blocked between the fibrillar chromatin and precondensed elongated chromosomal forms, and (e) the number and size of apoptotic bodies were inversely correlated with the progression of the cell cycle, with many small apoptotic bodies in early S phase and less and larger apoptotic bodies in late S phase.

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References

  1. Nelson WG, Kastan M. DNA strand breaks-the DNA template alteration that triggers p53 dependent DNA damage response pathways. Mol Cell Biol 1994; 14: 1815–1823.

    Google Scholar 

  2. Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW. Participation of p53 protein in the cellular response to DNA damage. Cancer Res 1991; 51: 6304–6311.

    Google Scholar 

  3. Kuerbitz SJ, Plunkett BS, Walsh WV, Kastan MB. Wild-type p53 is a cell cycle checkpoint determinant following irradiation. Proc Natl Acad Sci USA 1992; 89: 7491–7495.

    Google Scholar 

  4. Hall PA, McKee PH, Menage HD-P, Dover R, Lane DP. High levels of p53 protein in UV-irradiated normal human skin. Oncogene 1993; 8: 203–207.

    Google Scholar 

  5. Lu X, Lane DP. Differential induction of transcriptionally active p53 following UV or ionizing irradiation: Defects in chromosome instability syndromes? Cell 1993; 75: 765–778.

    Google Scholar 

  6. Yamaizumi M, Sugano T. UV-induced nuclear accumulation of p53 is evoked through DNA damage of actively transcribed genes independent of the cell cycle. Oncogene 1994; 9: 2775–2784.

    Google Scholar 

  7. Zhan Q, Carrier F, Fornace AJ. Induction of cellular p53 activity by DNA damaging agents and growth arrest. Mol Cell Biol 1993; 59(13): 4242–4250.

    Google Scholar 

  8. Tishler RB, Calderwood SK, Coleman CN, Price BD. Increases in sequence specific DNA binding by p53 Following treatment with chemotherapeutic and DNA damaging agents. Cancer Res 1993; 53: 2212–2216.

    Google Scholar 

  9. An WG, Kanekal M, Simon MC, Maltepe E, Blagosklony MV, Neckers LM. Stabilization of wild-type p53 by hypoxia-inducible factor 1 alpha. Nature 1998; 392: 405–408.

    Google Scholar 

  10. Graeber T, Osmanian C, Jacks T, et al. Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature 1996; 379: 88–91.

    Google Scholar 

  11. Gottlieb TM, Oren M. p53 and apoptosis. Seminars in Cancer Biology 1998; 8: 359–368.

    Google Scholar 

  12. Schwartz D, Rotter V. p53-dependent cell cycle control: Response to genotoxic stress. Semin Cancer Biol 1998; 8: 325–336.

    Google Scholar 

  13. Offer H, Zurer I, Banfalvi G, et al. p53 modulates base excision activity in a cell cycle-specific manner after genotoxic stress. Cancer Res 2001; 61: 88–96.

    Google Scholar 

  14. Adolph KW, Cheng SM, Laemmli UK. Role of nonhistone proteins in metaphase chromosome structure. Cell 1977; 12: 805–816.

    Google Scholar 

  15. Paulson JR, Laemmli UK. The structure of histone-depleted metaphase chromosomes. Cell 1977; 12: 817–828.

    Google Scholar 

  16. Benyajati C, Worcel A. Isolation, characterization and structure of the folded interphase genome of Drosophila melanogaster. Cell 1976; 9: 393–400.

    Google Scholar 

  17. Cook PR, Brazell IA. Mapping sequences in loops of nuclear DNA by their progressive detachment from the nuclear cage. Nucleic Acids Res 1980; 8: 2895–2907.

    Google Scholar 

  18. Earnshaw WC, Laemmli UK. Architecture of metaphase chromosomes and chromosome scaffolds. J Cell Biol 1983; 96: 84–93.

    Google Scholar 

  19. Rattner JB, Lin CC. Radial loops and helical coils coexist in metaphase chromosomes. Cell 1985; 41: 291–296.

    Google Scholar 

  20. Earnshaw WC. Mitotic chromosome structure. BioEssays 1988; 9: 147–150.

    Google Scholar 

  21. Wan KM, Nickerson JA, Krockmalnik G, Penman S. The nuclear matrix prepared by amine modification. Proc Natl Acad Sci USA 1999; 96: 933–938.

    Google Scholar 

  22. Comings DE. Arrangement of chromatin in the nucleus. Hum Genet 1980; 53: 131–143.

    Google Scholar 

  23. Cremer C, Cremer T, Zorn C, Cioreanu V. Partial irradiation of Chinese hamster cell nuclei and detection of unscheduled DNA synthesis in interphase and metaphase. A tool to investigate the arrangement of interphase chromosomes in mammalian cells. Hoppe Seyler's Z Physiol Chem 1979; 360: 244–245.

    Google Scholar 

  24. Sperling K, Luedtke EK. Arrangement of prematurely condensed chromosomes in cultured cells and lymphocytes of the Indian muntjac. Chromosoma 1981; 83: 541–553.

    Google Scholar 

  25. Xue Y, Wong J, Moreno, Young GT, Cote MK, Wang JW. NURD, a novel complex with both ATP-dependent chromatin-remodeling and histone deacetylase activities. Mol Cell 1998; 2: 851–861.

    Google Scholar 

  26. deCampos-Vidal B, Russo J, Mello ML. DNA content and chromatine texture of benzo[α]pyrene-transformed human breast epithelial cells as assessed by image analysis. Exp Cell Res 1998; 244: 77–82.

    Google Scholar 

  27. Juan G, Traganos F, James WM, et al. Histone H3 phosphorylation and expression of cyclins A and B1 measured in individual cells during their progression through G2 and mitosis. Cytometry 1998; 32: 71–77.

    Google Scholar 

  28. Heslop-Harrison JS, Huelskamp M, Wendroth S, Atkinson MD, Leicht AR, Benett MD. Chromatin and centromeric structures in interphase nuclei. In Brandham PE. ed. New Chromosome Conference III. London: Allan & Unwin 1988: 209–217.

    Google Scholar 

  29. Lemke J, Claussen J, Michel S, et al. The DNA-Based Structure of Human Chromosome 5 in Interphase. Am J Hum Genet 2002; 71: 1051–1059.

    Google Scholar 

  30. Banfalvi G, Sooki-Toth A, Sarkar N, Csuzi S, Antoni F. Nascent DNA synthesized reversibly permeable cells of mouse thymocytes. Eur J Biochem 1984; 139: 553–559.

    Google Scholar 

  31. Banfalvi G. Fluorescent analysis of replication and intermediates of chromatin folding in nuclei of mammalian cells. In Bach P, Reynolds CH, Clark JM, Mottley J, Poole PL, eds. Biotechnology Applications of Microinjection, Microscopic Imaging, and Fluorescence. New York, London: Plenum Press 1993: 111–119.

    Google Scholar 

  32. Aloni-Grinstein R, Zan-Bar I, Alboum I, Goldfinger N, Rotter V. Wild type p53 functions as a control protein in the differentiation pathway of the B-cell lineage. Oncogene 1993; 8: 3297–3305.

    Google Scholar 

  33. Basnakian AG, Banfalvi G, Sarkar N. Contribution of DNA polymerase δ to DNA replication in permeable CHO cells within nuclei of Chinese hamster ovary cells synchronized in S phase. Nucleic Acid Res 1989; 17: 4757–4767.

    Google Scholar 

  34. Pflumm MF. The role of DNA replication in chromosome condensation. Bioessays 2002; 24: 411–418.

    Google Scholar 

  35. Mascetti G, Carrara S, Vergani L. Relationship between chromatin compactness and dye uptake for in situ chromatin stained with DAPI. Cytometry 2001; 44: 113–119.

    Google Scholar 

  36. Mikhailov A, Rieder CL. Cell cycle stressed out of mitosis. Curr Biol 2002; 12: 331–333.

    Google Scholar 

  37. Iwano M, Fukui K, Tkaichi S, Isogai A. Globular fibrous structure in barley chromosomes revealed by high-resolution scanning electron microscopy. Chromosome Res 1997; 5: 341–349.

    Google Scholar 

  38. Sarkar A, Eroglu S, Poirier MG, Gupta P, Nemani A, Marko JF. Dynamics of chromosome compaction during mitosis. Exp Cell Res 2002; 277: 48–56.

    Google Scholar 

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Authors and Affiliations

  1. Department of Animal Anatomy and Physiology, University of Debrecen, Debrecen, 4032, Hungary

    G. Nagy, M. Gacsi, M. Klaisz & G. Banfalvi

  2. Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka, 900 28, Slovakia

    M. Rehak

  3. Division of Nephrology, University of Arkansas for Medical Sciences, Little Rock, AR, 72205, USA

    A. G. Basnakian

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  1. G. Nagy
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  2. M. Gacsi
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  4. A. G. Basnakian
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  5. M. Klaisz
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  6. G. Banfalvi
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Nagy, G., Gacsi, M., Rehak, M. et al. Gamma irradiation-induced apoptosis in murine pre-B cells prevents the condensation of fibrillar chromatin in early S phase. Apoptosis 9, 765–776 (2004). https://doi.org/10.1023/B:APPT.0000045790.86905.00

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  • DOI: https://doi.org/10.1023/B:APPT.0000045790.86905.00

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