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Non-histone chromosomal proteins. Their isolation and role in determining specificity of transcription in vitro.

Authors
  • Blüthmann, H
  • Mrozek, S
  • Gierer, A
Type
Published Article
Journal
European journal of biochemistry / FEBS
Publication Date
Oct 15, 1975
Volume
58
Issue
2
Pages
315–326
Identifiers
PMID: 1237403
Source
Medline
License
Unknown

Abstract

We describe a method for fractionation of chromatin components by selective dissociation with salt in buffers containing 5 M urea in combination with cromatography on hydroxyapatite at 4 degrees C. This results in two histone and four non-histone fractions which are recovered in high yield and with minimal proteolytic contamination. Template capacity measurements of the isolated chromatins and pre-saturation competition hybridization experiments support the idea that a group of non-histone proteins activate the transcription of specific DNA sequences which were not transcribed from purified DNA to the same extent. In reconstitution experiments a non-histone protein fraction, NH4, prepared from lymphocyte chromatin by hydroxyapatite chromatography is shown to cause transcription in vitro of lymphocyte-specific RNA sequences. A subfraction with a molecular weight of 30 000 comprising 40% of the NH4 fraction protein is characteristic for this tissue and not found in liver chromatin.

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