Fractionation of micrococcal nuclease-digested chromatin solubilized at physiologic ionic strength.
When mouse brain nuclei are optimally digested with micrococcal nuclease, most of the chromatin is soluble in a 180 mM salt/1 mM EDTA buffer [1]. At this ionic concentration, chromatin maintains its native structure [2]. In an attempt to selectively extract different fractions of chromatin from digested nuclei, we have examined the differential solubility of chromatin in the 180 mM salt buffer containing concentrations of MgCl2 ranging from 2 to 0 mM. The results suggest that digested chromatin may be fractionated into specific soluble chromatin fractions which correspond to nuclease-sensitive chromatin, bulk chromatin, and heterochromatin. These soluble fractions have a high molecular weight (up to 20 kbp), and contain a full complement of histones as well as a complex assortment of non-histone proteins. The residual insoluble fraction may be equivalent to a native, nuclear matrix-bound chromatin fraction.
D000818:Animals; D002467:Cell Nucleus / Q000648:ultrastructure; D005591:Chemical Fractionation; D002843:Chromatin / Q000032:analysis / Q000648:ultrastructure; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis; D004247:DNA / Q000032:analysis; D004276:DNA, Satellite / Q000032:analysis; D006570:Heterochromatin / Q000032:analysis; D006657:Histones / Q000032:analysis; D008274:Magnesium; D015636:Magnesium Chloride; D051379:Mice; D008836:Micrococcal Nuclease / Q000378:metabolism; D008970:Molecular Weight; D009994:Osmolar Concentration; D012995:Solubility
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004276:DNA, Satellite; D006570:Heterochromatin; D006657:Histones; D015636:Magnesium Chloride; D004247:DNA; D008836:Micrococcal Nuclease; D008274:Magnesium