The absence of oligonucleosomal DNA fragmentation during apoptosis of IMR-5 neuroblastoma cells: disappearance of the caspase-activated DNase.
Caspase-activated DNase is responsible for the oligonucleosomal DNA degradation during apoptosis. DNA degradation is thought to be important for multicellular organisms to prevent oncogenic transformation or as a mechanism of viral defense. It has been reported that certain cells, including some neuroblastoma cell lines such as IMR-5, enter apoptosis without digesting DNA in such a way. We have analyzed the causes for the absence of DNA laddering in staurosporine-treated IMR-5 cells, and we have found that most of the molecular mechanisms controlling apoptosis are well preserved in this cell line. These include degradation of substrates for caspases, blockade of cell death by antiapoptotic genes such as Bcl-2 or Bcl-X(L), or normal levels and adequate activation of caspase-3. Moreover, these cells display normal levels of caspase-activated DNase and its inhibitory protein, inhibitor of caspase-activated DNase, and their cDNA sequences are identical to those reported previously. Nevertheless, IMR-5 cells lose caspase-activated DNase during apoptosis and recover their ability to degrade DNA when human recombinant caspase-activated DNase is overexpressed. Our results lead to the conclusion that caspase-activated DNase is processed during apoptosis of IMR-5 cells, making these cells a good model to study the relevance of this endonuclease in physiological or pathological conditions.
Yuste
V J
VJ
Grup de Neurobiologia Molecular, Departament de Ciències Mèdiques Bàsiques, Universitat de Lleida, 25198 Lleida, Spain.
Bayascas
J R
JR
Llecha
N
N
Sánchez-López
I
I
Boix
J
J
Comella
J X
JX
D017209:Apoptosis; D001483:Base Sequence; D002843:Chromatin / Q000378:metabolism; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D009447:Neuroblastoma / Q000473:pathology*; D009707:Nucleosomes / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D002843:Chromatin; D017931:DNA Primers; D004273:DNA, Neoplasm; D009707:Nucleosomes; D003851:Deoxyribonucleases; C109663:caspase-activated deoxyribonuclease