title: A general method for purification of H1 histones that are active for repression of basal RNA polymerase II transcription.
abstract: H1 histones were purified from extracts of salt-treated nuclei as a co-product of RNA polymerase II transcription factors from both Drosophila embryos and HeLa cells by a simple and general method. This procedure was also used to purify H1 as co-product of the core histones from calf thymus. The key steps in this purification exploit the solubility of H1 in 2.26 M ammonium sulfate and the chromatographic properties of the highly charged H1 molecules on a phenyl-Sepharose resin. H1 that is prepared by this procedure is active for in vitro repression of basal RNA polymerase II transcription. This method provides a new means of purifying H1 by a mild procedure that is likely to be generally useful for studies of transcription and chromatin structure.
authors.affiliation: Department of Biology, University of California, San Diego, La Jolla 92093.
authors.forename: G E
authors.initials: GE
authors.lastname: Croston
authors.forename: L M
authors.initials: LM
authors.lastname: Lira
authors.forename: J T
authors.initials: JT
authors.lastname: Kadonaga
mesh_terms: D000645:Ammonium Sulfate; D000818:Animals; D002417:Cattle; D002847:Chromatography, Agarose; D004331:Drosophila melanogaster; D006367:HeLa Cells; D006657:Histones / Q000302:isolation & purification* / Q000494:pharmacology*; D006801:Humans; D012319:RNA Polymerase II / Q000235:genetics*; D012685:Sepharose / Q000031:analogs & derivatives; D013950:Thymus Gland / Q000737:chemistry; D014158:Transcription, Genetic / Q000187:drug effects*
chemical_list: D006657:Histones; C020927:phenyl-sepharose; D012685:Sepharose; D012319:RNA Polymerase II; D000645:Ammonium Sulfate