Role of alkaline endonucleases in the release of soluble chromatin from thymus, spleen and liver nuclei of normal and irradiated mice.
Thymus, spleen and liver nuclei released a large fraction of soluble chromatin in vitro when incubation was carried out in sucrose media containing low concentrations of CaCl2 and/or MgCl2. A significant fraction of deoxyribopolynucleotides (DPN) was also extracted from nuclei. After 30 min of incubation at 37 degrees C, the maximum release of soluble chromatin was observed near a pH of 8, which corresponds to the optimum pH of the alkaline endonuclease activity from thymus, spleen and liver. The soluble chromatin and DPN were precipitated by increasing the bivalent ion concentration of the medium. The protein/DNA ratio and the molecular weight of DNA suggest that the soluble chromatin and DPN represent nucleosome-like particles. The release of soluble chromatin in the first 4 hours of incubation was significantly increased if the nuclear fraction was isolated from the thymus and spleen of whole-body irradiated mice (1000 rad). This effect was absent in the liver nuclei.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002467:Cell Nucleus / Q000201:enzymology*; D002843:Chromatin / Q000378:metabolism*; D003037:Cobalt Radioisotopes; D004247:DNA / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism / Q000528:radiation effects*; D005260:Female; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology / Q000528:radiation effects* / Q000648:ultrastructure; D008274:Magnesium / Q000494:pharmacology; D051379:Mice; D009243:NAD / Q000378:metabolism; D013154:Spleen / Q000201:enzymology / Q000528:radiation effects* / Q000648:ultrastructure; D013950:Thymus Gland / Q000201:enzymology / Q000528:radiation effects* / Q000648:ultrastructure
D002843:Chromatin; D003037:Cobalt Radioisotopes; D009243:NAD; D004247:DNA; D004720:Endonucleases; D008274:Magnesium; D002118:Calcium