A Mg(2+)-dependent endonuclease is responsible for internucleosomal DNA fragmentation in human B lymphoblastic IM9 cells.
We have identified a Mg(2+)-dependent endonuclease activity from human B lymphoblastic IM9 cell lysates and nuclei using autodigestion method and DNA-native-polyacrylamide gel electrophoresis (DNA-native-PAGE) nuclease assay system. The level of the endonuclease activity in cell lysates was significantly decreased at certain stage by treatment of the cells with cycloheximide. However, the enzyme activity consistently remained for over 12 hours in the isolated nuclei of the apoptotic IM9 cells. The Mg(2+)-dependent endonuclease isolated from the nuclei by native-PAGE elution was able to catalyze the conversion of supercoiled plasmid DNA into linear form. This particular endonuclease activity was not detected in cycloheximide treated-U937 cells. Several lines of experimental evidence suggest that the Mg(2+)-dependent endonuclease localized in the nucleus may be responsible for the DNA fragmentation of apoptotic IM9 cells.
Kwon
H J
HJ
Department of Biochemistry, College of Science, and Bioproducts Research Center, Yonsei University, Seoul, Korea.
Kim
D S
DS
D017209:Apoptosis / Q000187:drug effects; D001402:B-Lymphocytes; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology*; D003513:Cycloheximide / Q000494:pharmacology; D053938:DNA Fragmentation; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009707:Nucleosomes / Q000502:physiology*; D010957:Plasmids; D013997:Time Factors; D020298:U937 Cells
D009707:Nucleosomes; D003513:Cycloheximide; D004720:Endonucleases