N-Butyrate incubation of immature chicken erythrocytes preferentially enhances the solubility of beta A chromatin

Abstract

The solubility of adult beta-globin chromatin (beta A chromatin) from immature chicken red blood cells can be controlled by the presence or absence of n-butyrate in a cell incubation medium. In the absence of n-butyrate, only a small percentage (approximately 4%) of the total beta A chromatin is in a soluble chromatin fraction following micrococcal nuclease digestion and centrifugation. This percentage increases to approximately 40-45% of the beta A chromatin if cells are incubated 1 hour in the presence of 10 mM sodium n-butyrate. The highest yield and enrichment of solubilized beta A chromatin is attained when 1-4% of the DNA is rendered acid soluble, and in buffers containing 1.5 - 5 mM MgCl2. The soluble beta A nucleohistone is nucleosome oligomer size (contains DNA 250-600 bases in length) and can be separated from soluble, transcriptionally inert mononucleosomes by agarose A-5m exclusion chromatography. The enhanced solubility appears to be specific for transcriptionally active chromatin. Whereas 40-45% of the beta A chromatin is recovered in the supernatant fraction from n-butyrate incubated immature erythrocytes, nucleohistone containing ovalbumin DNA sequences remains insoluble.