Ca2+/Mg2+-dependent endonuclease activation is an early event in VP-16-induced apoptosis of human breast cancer MCF7 cells in vitro

Irina A. Sokolova; Kenneth H. Cowan; Erasmus Schneider

doi:10.1016/0167-4889(94)00233-5

Ca²⁺/Mg²⁺-dependent endonuclease activation is an early event in VP-16-induced apoptosis of human breast cancer MCF7 cells in vitro

Irina A. Sokolova, Kenneth H. Cowan, Erasmus Schneider

Research output: Contribution to journal › Article › peer-review

38 Scopus citations

Abstract

Apoptosis is now recognized as one of the major processes regulating the size of cell populations. However, despite intensive investigations the biochemical and enzymological mechanisms involved in apoptosis remain unclear. In the present study we demonstrate activation of a Ca²⁺/Mg²⁺-dependent endonuclease during VP-16-induced apoptosis in MCF7 cells. Nuclease activation occurred prior to the appearance of internucleosomal DNA fragmentation, suggesting that this activation may be an early and possibly critical step in drug-induced apoptosis. Analysis of the internucleosomal DNA fragments showed that they contained phosphorylated 5′-ends indicating that they were produced by a Ca²⁺/Mg²⁺-dependent endonuclease.

Original language	English (US)
Pages (from-to)	135-142
Number of pages	8
Journal	Biochimica et Biophysica Acta - Molecular Cell Research
Volume	1266
Issue number	2
DOIs	https://doi.org/10.1016/0167-4889(94)00233-5
State	Published - 1995
Externally published	Yes

Keywords

10 mM Tris-HCl/1 m1vl EDTA (pH 8.0)
40 mM Tris-acetate/2 mM EDTA (pH 7.8)
4′,6-diamino-2-phenylindole
5-FdUrd
5-fluoro-2′-deoxyuridine
Apoptosis
CIAP
DAPI
DNA fragmentation
Endonuclease
PBS
SDS
TAE buffer
TE buffer
VP-16
calf intestinal alkaline phosphatase
etoposide
phosphate buffer solution
sodium dodecyl sulfate

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Access to Document

10.1016/0167-4889(94)00233-5

Cite this

@article{0491f42103914de9b38043944aeae41b,

title = "Ca2+/Mg2+-dependent endonuclease activation is an early event in VP-16-induced apoptosis of human breast cancer MCF7 cells in vitro",

abstract = "Apoptosis is now recognized as one of the major processes regulating the size of cell populations. However, despite intensive investigations the biochemical and enzymological mechanisms involved in apoptosis remain unclear. In the present study we demonstrate activation of a Ca2+/Mg2+-dependent endonuclease during VP-16-induced apoptosis in MCF7 cells. Nuclease activation occurred prior to the appearance of internucleosomal DNA fragmentation, suggesting that this activation may be an early and possibly critical step in drug-induced apoptosis. Analysis of the internucleosomal DNA fragments showed that they contained phosphorylated 5′-ends indicating that they were produced by a Ca2+/Mg2+-dependent endonuclease.",

keywords = "10 mM Tris-HCl/1 m1vl EDTA (pH 8.0), 40 mM Tris-acetate/2 mM EDTA (pH 7.8), 4′,6-diamino-2-phenylindole, 5-FdUrd, 5-fluoro-2′-deoxyuridine, Apoptosis, CIAP, DAPI, DNA fragmentation, Endonuclease, PBS, SDS, TAE buffer, TE buffer, VP-16, calf intestinal alkaline phosphatase, etoposide, phosphate buffer solution, sodium dodecyl sulfate",

author = "Sokolova, {Irina A.} and Cowan, {Kenneth H.} and Erasmus Schneider",

year = "1995",

doi = "10.1016/0167-4889(94)00233-5",

language = "English (US)",

volume = "1266",

pages = "135--142",

journal = "Biochimica et Biophysica Acta - Molecular Cell Research",

issn = "0167-4889",

publisher = "Elsevier",

number = "2",

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TY - JOUR

T1 - Ca2+/Mg2+-dependent endonuclease activation is an early event in VP-16-induced apoptosis of human breast cancer MCF7 cells in vitro

AU - Sokolova, Irina A.

AU - Cowan, Kenneth H.

AU - Schneider, Erasmus

PY - 1995

Y1 - 1995

N2 - Apoptosis is now recognized as one of the major processes regulating the size of cell populations. However, despite intensive investigations the biochemical and enzymological mechanisms involved in apoptosis remain unclear. In the present study we demonstrate activation of a Ca2+/Mg2+-dependent endonuclease during VP-16-induced apoptosis in MCF7 cells. Nuclease activation occurred prior to the appearance of internucleosomal DNA fragmentation, suggesting that this activation may be an early and possibly critical step in drug-induced apoptosis. Analysis of the internucleosomal DNA fragments showed that they contained phosphorylated 5′-ends indicating that they were produced by a Ca2+/Mg2+-dependent endonuclease.

AB - Apoptosis is now recognized as one of the major processes regulating the size of cell populations. However, despite intensive investigations the biochemical and enzymological mechanisms involved in apoptosis remain unclear. In the present study we demonstrate activation of a Ca2+/Mg2+-dependent endonuclease during VP-16-induced apoptosis in MCF7 cells. Nuclease activation occurred prior to the appearance of internucleosomal DNA fragmentation, suggesting that this activation may be an early and possibly critical step in drug-induced apoptosis. Analysis of the internucleosomal DNA fragments showed that they contained phosphorylated 5′-ends indicating that they were produced by a Ca2+/Mg2+-dependent endonuclease.

KW - 10 mM Tris-HCl/1 m1vl EDTA (pH 8.0)

KW - 40 mM Tris-acetate/2 mM EDTA (pH 7.8)

KW - 4′,6-diamino-2-phenylindole

KW - 5-FdUrd

KW - 5-fluoro-2′-deoxyuridine

KW - Apoptosis

KW - CIAP

KW - DAPI

KW - DNA fragmentation

KW - Endonuclease

KW - PBS

KW - SDS

KW - TAE buffer

KW - TE buffer

KW - VP-16

KW - calf intestinal alkaline phosphatase

KW - etoposide

KW - phosphate buffer solution

KW - sodium dodecyl sulfate

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DO - 10.1016/0167-4889(94)00233-5

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AN - SCOPUS:0029653416

SN - 0167-4889

VL - 1266

SP - 135

EP - 142

JO - Biochimica et Biophysica Acta - Molecular Cell Research

JF - Biochimica et Biophysica Acta - Molecular Cell Research

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