Cytoplasmic origin of the so-called nuclear neutral histone protease.
1. We have investigated the origin of proteolytic activity which causes degradation of histones in chromatin isolated from Xenopus liver and the rat liver at neutral pH. Polyacrylamide disc gel electrophoresis was used for detection of proteolytic products of histones. 2. No proteolytic degradation of histones occurs in chromatin isolated from Xenopus erythrocytes and rat liver according to our procedure even after prolonged incubation at pH 8.0 and pH 5.0. However with chromatin isolated from Xenopus liver a high level of histone degradation is observed under similar conditions. 3. Mixing isolated nuclei from Xenopus erythrocytes with a crude cytoplasmic fraction from Xenopus liver causes histone proteolysis in isolated chromatin at pH 8.0. In similar experiments with corresponding fractions from rat liver histone proteolysis can be introduced only after repeated freezing and thawing of the cytoplasmic fraction. 4. A purified lysosomal preparation from rat liver causes a similar type of histone degradation upon incubation with chromatin from Xenopus erythrocytes and rat liver. 5. The neutral proteolytic activity that can be introduced in isolated chromatin by a crude cytoplasmic fraction and by a purified lysosomal erythrocytes and rat liver. 5. The neutral proteolytic activity that can be introduced in isolated chromatin by a crude cytoplasmic fraction and by a purified lysosomal fraction from rat liver is inhibited by sodium bisulphite. 6. We conclude that the neutral proteolytic activity which causes degradation of histones in isolated chromatin is due to a contamination with neutral protease(s) originating from cytoplasmic organelles.
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology; D002843:Chromatin; D003593:Cytoplasm / Q000201:enzymology; D004912:Erythrocytes / Q000201:enzymology; D006657:Histones; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology; D010447:Peptide Hydrolases / Q000032:analysis*; D051381:Rats; D014981:Xenopus
D002843:Chromatin; D006657:Histones; D010447:Peptide Hydrolases