MAPPING AND POSITIONING DNA-BINDING PROTEINS ALONG GENOMIC DNA - STRUCTURE OF DROSOPHILA-MELANOGASTER RIBOSOMAL ALU-REPEATS AND 1.688 SATELLITE CHROMATIN

被引:18
作者
BELIKOV, SV [1 ]
BELGOVSKY, AI [1 ]
PARTOLINA, MP [1 ]
KARPOV, VL [1 ]
MIRZABEKOV, AD [1 ]
机构
[1] WA ENGELHARDT INST MOLEC BIOL, MOSCOW 117984, RUSSIA
关键词
D O I
10.1093/nar/21.20.4796
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 nonhistone protein contacts on satellite DNA. These data indicate precise positioning of non-histone proteins, histone Hl 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 succesive mapping and positioning proteins on genomic DNA.
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页码:4796 / 4802
页数:7
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