European Biophysics Journal

, 38:729

Chromosome shattering: a mitotic catastrophe due to chromosome condensation failure

Authors

  • B. Hübner
    • Department Biology II (Anthropology and Human Genetics)LMU Biozentrum
  • H. Strickfaden
    • Department Biology II (Anthropology and Human Genetics)LMU Biozentrum
  • S. Müller
    • Institute of Human GeneticsUniversity of Munich
    • Department Biology II (Anthropology and Human Genetics)LMU Biozentrum
    • Department Biology II (Anthropology and Human Genetics)LMU Biozentrum
    • Munich Center for Integrated Protein Science Munich (CIPSM)
Original Paper

DOI: 10.1007/s00249-009-0496-z

Cite this article as:
Hübner, B., Strickfaden, H., Müller, S. et al. Eur Biophys J (2009) 38: 729. doi:10.1007/s00249-009-0496-z

Abstract

Chromosome shattering has been described as a special form of mitotic catastrophe, which occurs in cells with unrepaired DNA damage. The shattered chromosome phenotype was detected after application of a methanol/acetic acid (MAA) fixation protocol routinely used for the preparation of metaphase spreads. The corresponding phenotype in the living cell and the mechanism leading to this mitotic catastrophe have remained speculative so far. In the present study, we used V79 Chinese hamster cells, stably transfected with histone H2BmRFP for live-cell observations, and induced generalized chromosome shattering (GCS) by the synergistic effect of UV irradiation and caffeine posttreatment. We demonstrate that GCS can be derived from abnormal mitotic cells with a parachute-like chromatin configuration (PALCC) consisting of a bulky chromatin mass and extended chromatin fibers that tether centromeres at a remote, yet normally shaped spindle apparatus. This result hints at a chromosome condensation failure, yielding a “shattered” chromosome complement after MAA fixation. Live mitotic cells with PALCCs proceeded to interphase within a period similar to normal mitotic cells but did not divide. Instead they formed cells with highly abnormal nuclear configurations subject to apoptosis after several hours. We propose a factor depletion model where a limited pool of proteins is involved both in DNA repair and chromatin condensation. Chromosome condensation failure occurs when this pool becomes depleted.

Keywords

Live-cell microscopyUV irradiation and DNA repairChromosome shatteringChromatin condensationMitotic catastropheFixation procedure

Abbreviations

FISH

Fluorescence in situ hybridization

GCS

Generalized chromosome shattering

H2BmRFP

Histone H2B conjugated to red fluorescent protein

H3pS10

Histone H3 with phosphorylated serine at position 10

MAA

Methanol/acetic acid

PALCC

Parachute-like chromatin configuration

PCC

Premature chromosome condensation

PCS

Partial chromosome shattering

PFA

Paraformaldehyde

SON

Supplemental online

Supplementary material

249_2009_496_MOESM1_ESM.tif (3 mb)
Fig. 12_SONa, b Three-color painting of Chinese hamster chromosomes 1 (green), 2 (blue), and 5 (red) in V79-H2BmRFP cells. a Apoptotic cells in PFA-fixed cultures after UV/caffeine treatment show a diffuse hybridization signal with strong intermingling of fluorescence signals reflecting a true fragmentation of chromosomes with a concomitant loss of the territorial structure. b Shadow-like chromatin patterns in MAA-fixed cultures lack any detectable hybridization signals. Scale bar = 5 μm Supplementary material 1 (tif 3043 kb)
249_2009_496_MOESM2_ESM.mpg (56.2 mb)
movie_1 Complete image sequence of the live-cell observation of V79-H2BmRFP control cells showing undisturbed cell proliferation up to 48 h. Images are taken each 15 min. The blurring after time point 1,860 min is due to a slight shifting of the z-stage after changing the medium. Picture sharpness in the late time points decreases due to the increasing cell density that hampers the autofocus function. The appearance of some apoptotic cells beyond time point 41 h is most probably due to the increased cell density. Scale bar = 10 µm Supplementary material 1 (MPG 57526 kb)
249_2009_496_MOESM3_ESM.mpg (13.3 mb)
movie_2 Complete image sequence of the live-cell observation of V79-H2BmRFP after UV irradiation/caffeine posttreatment. Scale bar = 10 µm Supplementary material 2 (MPG 13601 kb)

Copyright information

© European Biophysical Societies' Association 2009