Fractionation by micrococcal nuclease digestion of Drosophila embryo chromatin: isolation of a fraction enriched in two major nonhistone proteins.
Limited digestion of Drosophila melanogaster embryo nuclei with mitochondrial nuclease, followed by selective solubilization in 0.1 M NaCl, yields a soluble nucleoprotein fraction (S3) enriched in two dominant protein bands of apparent molecular weight of 44000 and 48000. The analysis of the nucleosome monomer and multimer peaks, separated on sucrose gradients after slight digestion with micrococcal nuclease, shows that these proteins are associated with chromatin subunits, and that they are principally found in the subnucleosome region of fraction S3. This doublet is tentatively identified as a member of the noncanonical HMG of Drosophila. The thermal denaturation and the circular dichroism spectra of the fraction soluble in 0.1 M NaCl (S3) and insoluble fraction (P3) show that both fractions are also structurally different.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000032:analysis*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000302:isolation & purification*; D002942:Circular Dichroism; D004331:Drosophila melanogaster / Q000032:analysis*; D004625:Embryo, Nonmammalian; D006609:High Mobility Group Proteins; D006358:Hot Temperature; D008836:Micrococcal Nuclease; D008970:Molecular Weight; D009707:Nucleosomes / Q000032:analysis*; D011489:Protein Denaturation
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006609:High Mobility Group Proteins; D009707:Nucleosomes; D008836:Micrococcal Nuclease