Abstract
Internucleosomal DNA fragmentation following the activation of endonucleases is the common end point of apoptosis. DNase I, a Ca 2+/Mg 2+-dependent endonuclease ubiquitously expressed in mammalian tissues, is believed to play a role in this process. To analyze the in vivo function of this enzyme in human cells, we have generated a cell line with targeted disruption of the DNase I gene, as well as several stable cell lines which overexpress the DNase I gene. Inactivation of the human DNase I gene was obtained in the Jurkat T cell clone JA3, characterized by high susceptibility to apoptotic cell death induced by pharmacological stimuli. JA3 cells, after disruption of the DNase I gene, became resistant to apoptotic stimuli. DNase I was overexpressed in the human cell lines JA3, K562 (erythroleukemia), M 14 (melanoma) and CEM (T cell lymphoma). Remarkably, stable overexpression of DNase I gene resulted in accelerated apoptosis in JA3 cells and induced apoptosis in K562, CEM and M14 cell lines, which are otherwise resistant to internucleosomal DNA degradation following pharmacological stimuli. Our study provides the first in vivo evidence that DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis.
| Original language | English |
|---|---|
| Pages (from-to) | 743-751 |
| Number of pages | 9 |
| Journal | European Journal of Immunology |
| Volume | 31 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2001 |
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Keywords
- Chemotherapy
- DNA fragmentation
- DNase I
- Gene targeting
ASJC Scopus subject areas
- Immunology
Cite this
DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis. / Oliveri, Mara; Daga, Antonio; Cantoni, Claudia; Lunardi, Claudio; Millo, Romano; Puccetti, Antonio.
In: European Journal of Immunology, Vol. 31, No. 3, 2001, p. 743-751.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis
AU - Oliveri, Mara
AU - Daga, Antonio
AU - Cantoni, Claudia
AU - Lunardi, Claudio
AU - Millo, Romano
AU - Puccetti, Antonio
PY - 2001
Y1 - 2001
N2 - Internucleosomal DNA fragmentation following the activation of endonucleases is the common end point of apoptosis. DNase I, a Ca 2+/Mg 2+-dependent endonuclease ubiquitously expressed in mammalian tissues, is believed to play a role in this process. To analyze the in vivo function of this enzyme in human cells, we have generated a cell line with targeted disruption of the DNase I gene, as well as several stable cell lines which overexpress the DNase I gene. Inactivation of the human DNase I gene was obtained in the Jurkat T cell clone JA3, characterized by high susceptibility to apoptotic cell death induced by pharmacological stimuli. JA3 cells, after disruption of the DNase I gene, became resistant to apoptotic stimuli. DNase I was overexpressed in the human cell lines JA3, K562 (erythroleukemia), M 14 (melanoma) and CEM (T cell lymphoma). Remarkably, stable overexpression of DNase I gene resulted in accelerated apoptosis in JA3 cells and induced apoptosis in K562, CEM and M14 cell lines, which are otherwise resistant to internucleosomal DNA degradation following pharmacological stimuli. Our study provides the first in vivo evidence that DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis.
AB - Internucleosomal DNA fragmentation following the activation of endonucleases is the common end point of apoptosis. DNase I, a Ca 2+/Mg 2+-dependent endonuclease ubiquitously expressed in mammalian tissues, is believed to play a role in this process. To analyze the in vivo function of this enzyme in human cells, we have generated a cell line with targeted disruption of the DNase I gene, as well as several stable cell lines which overexpress the DNase I gene. Inactivation of the human DNase I gene was obtained in the Jurkat T cell clone JA3, characterized by high susceptibility to apoptotic cell death induced by pharmacological stimuli. JA3 cells, after disruption of the DNase I gene, became resistant to apoptotic stimuli. DNase I was overexpressed in the human cell lines JA3, K562 (erythroleukemia), M 14 (melanoma) and CEM (T cell lymphoma). Remarkably, stable overexpression of DNase I gene resulted in accelerated apoptosis in JA3 cells and induced apoptosis in K562, CEM and M14 cell lines, which are otherwise resistant to internucleosomal DNA degradation following pharmacological stimuli. Our study provides the first in vivo evidence that DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis.
KW - Chemotherapy
KW - DNA fragmentation
KW - DNase I
KW - Gene targeting
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UR - http://www.scopus.com/inward/citedby.url?scp=0035081830&partnerID=8YFLogxK
U2 - 10.1002/1521-4141(200103)31:3<743::AID-IMMU743>3.0.CO;2-9
DO - 10.1002/1521-4141(200103)31:3<743::AID-IMMU743>3.0.CO;2-9
M3 - Article
C2 - 11241278
AN - SCOPUS:0035081830
VL - 31
SP - 743
EP - 751
JO - European Journal of Immunology
JF - European Journal of Immunology
SN - 0014-2980
IS - 3
ER -