[Effect of the conditions of isolation and incubation of the nuclei on digestion of DNA chromatin by calcium- and magnesium-dependent endonuclease. Change in the spectrum of chromosomal proteins and possible role of DNA-protein interactions].
Digesting of chromosomal DNA of interphase rat liver nuclei by Ca, Mg-dependent endonuclease in situ in the presence of chelating agents results in the appearance of the soluble DNP--up to 30% of the total DNA. In addition, 50% of the chromatin is solubilised after mild ultrasonication. In the absence of the chelating agents the degree of fragmentation is considerably increased. The process is accompanied by a loss of some histone and nonhistone chromosomal proteins; the nonhistone proteins are lost selectively. The preliminary removal of the nuclear membrane and significant part of the proteins by tritone X-100 promotes the chromatin degradation and the appearance of low molecular weight fragments. The DNA-fragments of solubilised chromatin are similar to the DNA-fragments of residual chromatin, but in the presence of the chelating agents the latter does not contain monomeric fragments.
D000818:Animals; D002118:Calcium; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000032:analysis / Q000378:metabolism*; D002875:Chromosomes / Q000378:metabolism; D003470:Culture Media; D046508:Culture Techniques; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D003852:Deoxyribonucleoproteins / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D008274:Magnesium; D008297:Male; D009698:Nucleoproteins / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats
D002843:Chromatin; D003470:Culture Media; D003852:Deoxyribonucleoproteins; D009698:Nucleoproteins; D011092:Polyethylene Glycols; D004247:DNA; D004720:Endonucleases; D008274:Magnesium; D002118:Calcium
Vliianie usloviĭ vydeleniia i inkubatsii iader na droblenie DNK khromatina Ca, Mg-zavisimoi endonukleazoĭ. Izmenenie spektra khromosomal'nykh belkov i vozmozhnaia rol' DNK-belkovykh vzaimodeistviĭ.