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Related Experiment Videos

Apoptotic nuclear morphological change without DNA fragmentation.

H Sakahira1, M Enari, Y Ohsawa

  • 1Department of Genetics, Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 2-2 Yamada-oka, Suita, Osaka, 565-0871, Japan.

Current Biology : CB
|May 26, 1999
PubMed
Summary
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Apoptosis involves DNA fragmentation and chromatin condensation. This study shows that caspase-activated DNase (CAD) causes DNA fragmentation, while chromatin condensation occurs independently of DNA degradation.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Apoptosis is a programmed cell death process.
  • Morphologically, apoptosis involves cell and nuclear condensation and fragmentation.
  • Biochemically, apoptosis is characterized by DNA fragmentation into nucleosomal units.

Purpose of the Study:

  • To investigate if nuclear morphological changes during apoptosis are caused by chromosomal DNA degradation.
  • To determine the role of caspase-activated DNase (CAD) in apoptosis-induced DNA fragmentation and chromatin condensation.

Main Methods:

  • Treatment of human T-cell lymphoma Jurkat cells and their caspase-resistant ICAD transformants with staurosporine.
  • Analysis of chromosomal DNA fragmentation and chromatin condensation patterns.

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Main Results:

  • Staurosporine induced nucleosomal DNA fragmentation and large-scale chromatin fragmentation (50-200 kb) in Jurkat cells.
  • In cells with caspase-resistant ICAD, chromosomal DNA remained intact, but chromatin condensed.
  • These findings suggest CAD is responsible for DNA fragmentation.

Conclusions:

  • Large-scale chromatin fragmentation and nucleosomal DNA fragmentation are mediated by an ICAD-inhibitable DNase, likely CAD.
  • Chromatin condensation during apoptosis is, at least partly, independent of chromosomal DNA degradation.