DNase I mediates internucleosomal DNA degradation in human cells undergoing drug-induced apoptosis

Mara Oliveri, Antonio Daga, Claudia Cantoni, Claudio Lunardi, Romano Millo, Antonio Puccetti

Research output: Contribution to journalArticlepeer-review

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 languageEnglish
Pages (from-to)743-751
Number of pages9
JournalEuropean Journal of Immunology
Volume31
Issue number3
DOIs
Publication statusPublished - 2001

Keywords

  • Chemotherapy
  • DNA fragmentation
  • DNase I
  • Gene targeting

ASJC Scopus subject areas

  • Immunology

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