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

Yuri A. Lazebnik, Susan Cole, Carol A. Cooke, William G Nelson, William C. Earnshaw

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)7-22
Number of pages16
JournalJournal of Cell Biology
Volume123
Issue number1
StatePublished - Oct 1993

Fingerprint

Systems Analysis
Apoptosis
Chickens
Nuclear Lamina
Cell-Free System
Nuclear Envelope
S Phase
Cell Division
Chromatin
Hepatocellular Carcinoma
In Vitro Techniques
DNA

ASJC Scopus subject areas

  • Cell Biology

Cite this

Nuclear events of apoptosis in vitro in cell-free mitotic extracts : A model system for analysis of the active phase of apoptosis. / Lazebnik, Yuri A.; Cole, Susan; Cooke, Carol A.; Nelson, William G; Earnshaw, William C.

In: Journal of Cell Biology, Vol. 123, No. 1, 10.1993, p. 7-22.

Research output: Contribution to journalArticle

Lazebnik, Yuri A. ; Cole, Susan ; Cooke, Carol A. ; Nelson, William G ; Earnshaw, William C. / Nuclear events of apoptosis in vitro in cell-free mitotic extracts : A model system for analysis of the active phase of apoptosis. In: Journal of Cell Biology. 1993 ; Vol. 123, No. 1. pp. 7-22.
@article{9060061c1b1745bc8088503b9e14d757,
title = "Nuclear events of apoptosis in vitro in cell-free mitotic extracts: A model system for analysis of the active phase of apoptosis",
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.",
author = "Lazebnik, {Yuri A.} and Susan Cole and Cooke, {Carol A.} and Nelson, {William G} and Earnshaw, {William C.}",
year = "1993",
month = "10",
language = "English (US)",
volume = "123",
pages = "7--22",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "1",

}

TY - JOUR

T1 - Nuclear events of apoptosis in vitro in cell-free mitotic extracts

T2 - A model system for analysis of the active phase of apoptosis

AU - Lazebnik, Yuri A.

AU - Cole, Susan

AU - Cooke, Carol A.

AU - Nelson, William G

AU - Earnshaw, William C.

PY - 1993/10

Y1 - 1993/10

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0027437541&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027437541&partnerID=8YFLogxK

M3 - Article

C2 - 8408207

AN - SCOPUS:0027437541

VL - 123

SP - 7

EP - 22

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 1

ER -