Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin.
Chromatin structure of so-called 'Alu-repeat' in D. melanogaster ribosomal non-transcribed spacer that contains sequences homologous to the promoter of ribosomal genes has been studied. Using the 'protein image' hybridization assay based on UV-light-induced DNA-protein crosslinking and 2-D gel retardation electrophoresis, two proteins of the molecular mass of 50 kD (rABP50) and 70 kD (rABP70), associated with 'Alu-repeat' DNA have been found. Exo III mapping of crosslinking sites and DNase I footprinting have provided a detailed map of H1, rABP50 and rABP70 contacts within the 'Alu-repeat' and H1 and a non-histone protein contacts on satellite DNA. These data indicate precise positioning of non-histone proteins, histone H1 and nucleosomes within genomic regions studied and account for the presence of unusual 240 bp long nucleosomal particles in 'Alu-repeats'. The same approach can be adapted for successive mapping and positioning proteins on genomic DNA.
Belikov
S V
SV
W.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.
Belgovsky
A I
AI
Partolina
M P
MP
Karpov
V L
VL
Mirzabekov
A D
AD
D000818:Animals; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004275:DNA, Ribosomal / Q000235:genetics*; D004276:DNA, Satellite / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics*; D006657:Histones / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism; D011485:Protein Binding; D012091:Repetitive Sequences, Nucleic Acid
D002843:Chromatin; D004275:DNA, Ribosomal; D004276:DNA, Satellite; D004268:DNA-Binding Proteins; D006657:Histones; D009707:Nucleosomes; D004247:DNA