Levinger L
J Biol Chem. 1985 Sep 25;260(21):11799-804.
Nucleosomes have been fractionated on nondenaturing polyacrylamide gels, and nucleosome subtypes containing the Drosophila melanogaster specific protein D1 and ubiquitinated core histone H2A were identified by solubility in 0.1 M NaCl before nucleoprotein gel electrophoresis. Nucleosomes which contain DNA complementary to the 1.672 density simple satellite (sequence-AATAT-) bind protein D1, as demonstrated by two-dimensional hybridization mapping. This hybridization pattern allows the identification of D1 dinucleosomes, which, like D1 mononucleosomes, are reduced in mobility on the first dimension (nucleoprotein) gel by the addition of D1, an AT sequence-specific DNA-binding protein. The 1.705 density simple satellite (sequence-AAGAG-) is also found in nucleosomes, in a radically different subset from those of the -AATAT- DNA sequence. -AAGAG- nucleosomes do not contain D1 protein, but appear to be enriched in ubiquitinated core histone H2A. One-dimensional hybridization patterns suggest that -AAGAG- nucleosomal DNA is rapidly trimmed to a shorter DNA length than either bulk or -AATAT- nucleosomes.
核小体已在非变性聚丙烯酰胺凝胶上进行了分级分离,并且通过在核蛋白凝胶电泳前在0.1M NaCl中的溶解性鉴定出了含有黑腹果蝇特异性蛋白D1和泛素化核心组蛋白H2A的核小体亚型。通过二维杂交图谱证明,含有与1.672密度简单卫星(序列 - AATAT - )互补的DNA的核小体结合蛋白D1。这种杂交模式允许鉴定D1二聚核小体,其与D1单核小体一样,通过添加D1(一种AT序列特异性DNA结合蛋白)在第一维(核蛋白)凝胶上的迁移率降低。在核小体中也发现了1.705密度简单卫星(序列 - AAGAG - ),它与 - AATAT - DNA序列的核小体属于完全不同的子集。 - AAGAG - 核小体不包含D1蛋白,但似乎富含泛素化核心组蛋白H2A。一维杂交模式表明, - AAGAG - 核小体DNA比整体或 - AATAT - 核小体更快地被修剪成更短的DNA长度。