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PMID: 8408207 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

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

The Journal of cell biology ·Vol. 123 ·No. 1 ·1993-10-00 ·Pages 7-22

Lazebnik YA, Cole S, Cooke CA, Nelson WG, Earnshaw WC

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.

MeSH Terms
Animals Aphidicolin/pharmacology Apoptosis/drug effects,physiology Cell Fractionation Cell Nucleus/drug effects,physiology,ultrastructure Cell-Free System Chickens Chromosomes DNA Damage/drug effects,physiology HeLa Cells Humans Mice Mitosis/physiology Nuclear Envelope/metabolism Nucleosomes/metabolism Protamine Kinase/metabolism S Phase Tumor Cells, Cultured Zinc/pharmacology
Chemicals
Nucleosomes Aphidicolin Protamine Kinase Zinc
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Lazebnik Y A
Department of Cell Biology, Johns Hopkins School of Medicine, Baltimore, Maryland 21205.
Cole S
Cooke C A
Nelson W G
Earnshaw W C
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.123.1.7
Published
1993-10-00
Pages
7-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2119808
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.123.1.7 PMC Full Text View journal details
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