Distribution of high mobility group proteins 1/2, E and 14/17 and linker histones H1 and H5 on transcribed and non-transcribed regions of chicken erythrocyte chromatin

Nucleic Acids Res. 1991 Feb 25;19(4):717-25. doi: 10.1093/nar/19.4.717.

Abstract

Quantitative analysis of distribution of chromosomal proteins on single copy DNA sequences has been further developed. Our approach consists of DNA-protein crosslinking within whole cells or isolated nuclei, specific immunoaffinity isolation of crosslinked complexes via protein and identification of crosslinked DNA by hybridisation with single-stranded DNA probes. The present study shows that transcribed chromatin of chicken embryonic erythrocyte beta globin gene is characterized by about 1.5-2.5-fold higher density of HMG 14/17 and 2-fold lower density of H1 and H5 as compared with non-transcribed chromatin of ovalbumin and lysozyme genes, whereas HMG 1/2, E proteins were equally distributed between DNA of both transcribed and non-transcribed genes. The depletion of H1/H5 in beta globin sequences was verified by the 'protein image' hybridisation technique (1). The DNase I hypersensitive site located 5' upstream from beta globin gene is deficient in all the proteins assayed, what implies a drastic disruption in the nucleosomal array. Minor quantitative changes of protein pattern suggest transient local perturbation of the chromatin on transcription.

MeSH terms

  • Animals
  • Blotting, Western
  • Chickens
  • Chromatin*
  • Cross-Linking Reagents
  • DNA Probes
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Erythrocytes / metabolism*
  • High Mobility Group Proteins / genetics*
  • Histones / genetics*
  • Muramidase / genetics
  • Nucleic Acid Hybridization
  • Ovalbumin / genetics
  • Precipitin Tests
  • Restriction Mapping
  • Transcription, Genetic*

Substances

  • Chromatin
  • Cross-Linking Reagents
  • DNA Probes
  • High Mobility Group Proteins
  • Histones
  • Ovalbumin
  • Muramidase