Detection of Mg(2+)-dependent endonuclease activity in myeloid leukemia cell nuclei capable of producing internucleosomal DNA cleavage.
We detected Mg(2+)-dependent, Ca(2+)-independent endonuclease activity in non-apoptotic myeloid leukemia cell nuclei using autodigestion method which cleaved the chromatin of the autologous leukemia cells to an oligonucleosomal length pattern. Similar endonuclease activity could be successfully recovered in the protein extracts of the human leukemia cell nuclei. The extracts consistently elicited characteristic DNA cleavage of another leukemia cell (KG-1) nuclei as the target, the enzyme activity of which had been inactivated. We propose that this method is a useful tool for the study of endonucleases involved in apoptosis.
Kawabata
H
H
Department of Medicine, Faculty of Medicine, Kyoto University, Japan.
Anzai
N
N
Masutani
H
H
Hirama
T
T
Yoshida
Y
Y
Okuma
M
M
D000818:Animals; D017209:Apoptosis; D002467:Cell Nucleus / Q000201:enzymology*; D004273:DNA, Neoplasm / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D006801:Humans; D007951:Leukemia, Myeloid; D008214:Lymphocytes / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology*; D008297:Male; D009687:Nuclear Proteins / Q000302:isolation & purification / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013601:T-Lymphocytes / Q000201:enzymology; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D009687:Nuclear Proteins; D009707:Nucleosomes; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I; C019329:deoxyribonuclease II; D008274:Magnesium