Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin.
Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin.
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沿着基因组 DNA 绘制和定位 DNA 结合蛋白。
DOI：
10.1093/nar/21.20.4796
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发表时间：
1993
影响因子：
14.9
通讯作者：
A. Mirzabekov
中科院分区：
文献类型：
--
作者：
S. Belikov;A. I. Belgovsky;M. Partolina;V. Karpov;A. Mirzabekov
已经研究了黑腹果蝇核糖体非转录间隔区中所谓“Alu-重复序列”的染色质结构，该间隔区含有与核糖体基因启动子同源的序列。使用基于紫外线诱导 DNA-蛋白质交联和 2-D 凝胶延迟电泳的“蛋白质图像”杂交测定，发现了与“Alu 重复”DNA 相关的两种分子量为 50 kD (rABP50) 和 70 kD (rABP70) 的蛋白质。 Exo III 交联位点图谱和 DNase I 足迹分析提供了“Alu 重复序列”内 H1、rABP50 和 rABP70 接触的详细图谱，以及卫星 DNA 上 H1 和非组蛋白接触的详细图谱。这些数据表明非组蛋白、组蛋白 H1 和核小体在所研究的基因组区域内的精确定位，并解释了“Alu 重复序列”中不寻常的 240 bp 长核小体颗粒的存在。相同的方法可以适用于在基因组 DNA 上连续绘制和定位蛋白质。
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.