Characterization of an Mg2+-dependent endonucleolytic activity of the rat hepatocyte nuclear matrix.
Initial degradation of chromatin into high-molecular mass DNA fragments during apoptosis reflects the periodicity of chromatin organization into nuclear matrix-attached loops. In this article, we put forward the hypothesis that this pattern of DNA cleavage is also a result of the localization of an endonuclease on the nuclear matrix. Namely, we observed an endonucleolytic activity of the isolated rat hepatocyte nuclear matrix. It was Mg2+-dependent, with an optimal activity at pH 7.2 in the absence of either Na+ or K+. It was fully active in the presence of Zn2+ and capable of introducing single-strand breaks into plasmid DNA. It did not display a sequence-specific activity. A 23 kDa DNA nuclease that was principally localized on the rat hepatocyte nuclear matrix was detected. The enzyme shared the biochemical requirements with the nuclear matrix endonucleolytic activity, thus we proposed that p23 could be responsible for the endonucleolytic activity of the nuclear matrix. In view of its properties and preferential localization on the nuclear matrix, the endonuclease described herein could be a possible candidate that brings about initial DNA cleavage during apoptosis.
Molecular Biology Laboratory, Institute for Biological Research, 29, Novembra 142, Belgrade 11060, Serbia and Montenegro.
Nevena
N
Grdović
Goran
G
Poznanović
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology*; D002843:Chromatin / Q000378:metabolism; D004249:DNA Damage / Q000502:physiology; D004720:Endonucleases / Q000378:metabolism*; D022781:Hepatocytes / Q000201:enzymology*; D007477:Ions / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism*; D008297:Male; D015530:Nuclear Matrix / Q000201:enzymology*; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000378:metabolism; D051381:Rats
D002843:Chromatin; D007477:Ions; D004720:Endonucleases; D008274:Magnesium