Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis

Lazebnik, Y. A., Cole, S., Cooke, C. A., Nelson, W. G., Earnshaw, W. C. (1993) Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis. Journal of Cell Biology, 123 (1). pp. 7-22. ISSN 0021-9525

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URL: http://www.ncbi.nlm.nih.gov/pubmed/8408207
DOI: 10.1083/jcb.123.1.7

Abstract

We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated nuclei. The system uses extracts prepared from mitotic chicken hepatoma cells following a sequential S phase/M phase synchronization. When nuclei are added to these extracts, the chromatin becomes highly condensed into spherical domains that ultimately extrude through the nuclear envelope, forming apoptotic bodies. The process is highly synchronous, and the structural changes are completed within 60 min. Coincident with these morphological changes, the nuclear DNA is cleaved into a nucleosomal ladder. Both processes are inhibited by Zn2+, an inhibitor of apoptosis in intact cells. Nuclear lamina disassembly accompanies these structural changes in added nuclei, and we show that lamina disassembly is a characteristic feature of apoptosis in intact cells of mouse, human and chicken. This system may provide a powerful means of dissecting the biochemical mechanisms underlying the final stages of apoptosis.

Item Type: Paper
Uncontrolled Keywords: Animals Aphidicolin Apoptosis Cell Fractionation Cell Nucleus Cell-Free System Chickens Chromosomes DNA Damage HeLa Cells Humans Mice Mitosis Nuclear Envelope Nucleosomes Protamine Kinase S Phase Tumor Cells, Cultured Zinc
Subjects: organs, tissues, organelles, cell types and functions > cell types and functions > cell functions > apoptosis
organs, tissues, organelles, cell types and functions > cell types and functions > cell functions
organs, tissues, organelles, cell types and functions > cell types and functions
organs, tissues, organelles, cell types and functions > organelles, types and functions > mitosis
organs, tissues, organelles, cell types and functions > organelles, types and functions
CSHL Authors:
Communities: CSHL labs > Labeznik lab
Depositing User: Matt Covey
Date: 1993
Date Deposited: 12 Dec 2012 17:32
Last Modified: 12 Dec 2012 17:32
PMCID: PMC2119808
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26405

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