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.
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.
Postnikov
Y V
YV
Engelhardt Institute of Molecular Biology, USSR Academy of Sciences, Moscow.
Shick
V V
VV
Belyavsky
A V
AV
Khrapko
K R
KR
Brodolin
K L
KL
Nikolskaya
T A
TA
Mirzabekov
A D
AD
D000818:Animals; D015153:Blotting, Western; D002645:Chickens; D002843:Chromatin; D003432:Cross-Linking Reagents; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000235:genetics*; D006657:Histones / Q000235:genetics*; D009113:Muramidase / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010047:Ovalbumin / Q000235:genetics; D011233:Precipitin Tests; D015183:Restriction Mapping; D014158:Transcription, Genetic
D002843:Chromatin; D003432:Cross-Linking Reagents; D015342:DNA Probes; D006609:High Mobility Group Proteins; D006657:Histones; D010047:Ovalbumin; D009113:Muramidase