Nucleosome disruption precedes transcription and is largely limited to the transcribed domain of globin genes in murine erythroleukemia cells.
We used micrococcal nuclease to separate murine erythroleukemia cell (MELC) chromatin into soluble and insoluble fractions which differ in gene content and chromatin structure. Genes that are not expressed in the erythroid lineage, such as the Ig alpha and albumin genes, distribute preferentially into the soluble rather than the insoluble fraction, and are organized into nucleosomes in both fractions. Both alpha 1- and beta maj-globin genes are enriched in the insoluble fraction and are organized into structures that are partially devoid of nucleosomes in uninduced MELC, when the genes are transcriptionally inactive. Following chemical induction of MELC and the onset of globin gene transcription, globin gene enrichment and nucleosome disruption in the insoluble chromatin fraction increase. Using seven DNA subclones that span the beta maj-globin gene we show that insolubility and nucleosome disruption are largely limited to DNA sequences lying within the transcribed domain. Non-transcribed, flanking sequences are soluble and organized into nucleosomes. In addition, the globin genes found in insoluble, non-nucleosomal chromatin contain previously engaged RNA polymerases which can elongate globin RNA chains in vitro in a pattern qualitatively and quantitatively similar to intact nuclei. These results are discussed in terms of a model for globin gene activation during erythropoeisis.
Cohen
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Sheffery
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D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005796:Genes; D005914:Globins / Q000235:genetics*; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D051379:Mice; D008836:Micrococcal Nuclease; D008957:Models, Genetic; D009693:Nucleic Acid Hybridization; D009707:Nucleosomes; D012313:RNA / Q000096:biosynthesis; D014158:Transcription, Genetic
D002843:Chromatin; D009707:Nucleosomes; D012313:RNA; D005914:Globins; D008836:Micrococcal Nuclease