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沿基因组DNA对DNA结合蛋白进行定位和映射。黑腹果蝇核糖体“Alu重复序列”和1.688卫星染色质的结构。

Mapping and positioning DNA-binding proteins along genomic DNA. Structure of D. melanogaster ribosomal 'Alu-repeats' and 1.688 satellite chromatin.

作者信息

Belikov S V, Belgovsky A I, Partolina M P, Karpov V L, Mirzabekov A D

机构信息

W.A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow.

出版信息

Nucleic Acids Res. 1993 Oct 11;21(20):4796-802. doi: 10.1093/nar/21.20.4796.

DOI:10.1093/nar/21.20.4796
PMID:8233828
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC331508/
Abstract

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.

摘要

对果蝇核糖体非转录间隔区中所谓的“Alu重复序列”的染色质结构进行了研究，该区域包含与核糖体基因启动子同源的序列。使用基于紫外线诱导的DNA-蛋白质交联和二维凝胶阻滞电泳的“蛋白质图像”杂交分析，发现了两种与“Alu重复序列”DNA相关的蛋白质，分子量分别为50 kD（rABP50）和70 kD（rABP70）。交联位点的外切核酸酶III图谱分析和DNase I足迹分析提供了“Alu重复序列”内H1、rABP50和rABP70接触位点以及卫星DNA上H1和一种非组蛋白接触位点的详细图谱。这些数据表明在所研究的基因组区域内非组蛋白、组蛋白H1和核小体的精确定位，并解释了“Alu重复序列”中存在异常的240 bp长核小体颗粒的原因。同样的方法可用于在基因组DNA上连续定位和测绘蛋白质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/b9353c4c7f9d/nar00069-0152-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/6ec27431b6e3/nar00069-0150-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/c956e1a53bc5/nar00069-0151-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/26cf74def235/nar00069-0151-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/a0a6c6bb147d/nar00069-0152-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/e9c1b036795b/nar00069-0152-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/b9353c4c7f9d/nar00069-0152-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/6ec27431b6e3/nar00069-0150-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/c956e1a53bc5/nar00069-0151-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/26cf74def235/nar00069-0151-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/a0a6c6bb147d/nar00069-0152-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/e9c1b036795b/nar00069-0152-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7501/331508/b9353c4c7f9d/nar00069-0152-c.jpg

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本文引用的文献

1
Two non-histone proteins are associated with the promoter region and histone HI with the transcribed region of active hsp-70 genes as revealed by UV-induced DNA-protein crosslinking in vivo.体内紫外线诱导的DNA-蛋白质交联显示，两种非组蛋白与活性hsp-70基因的启动子区域相关联，组蛋白H1与转录区域相关联。
Nucleic Acids Res. 1993 Feb 25;21(4):1031-4. doi: 10.1093/nar/21.4.1031.
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Primary organization of the nucleosome core particles. Sequential arrangement of histones along DNA.核小体核心颗粒的初级结构。组蛋白沿DNA的顺序排列。
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Repeated genes in eukaryotes.
真核生物中的重复基因。
Annu Rev Biochem. 1980;49:727-64. doi: 10.1146/annurev.bi.49.070180.003455.
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A mosaicism in the higher order structure of Xenopus oocyte nucleolar chromatin prior to and during ribosomal gene transcription.非洲爪蟾卵母细胞核仁染色质在核糖体基因转录之前及转录期间的高阶结构中的镶嵌性。
Cell. 1981 Mar;23(3):711-20. doi: 10.1016/0092-8674(81)90434-7.
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Nontranscribed spacer sequences promote in vitro transcription of Drosophila ribosomal DNA.非转录间隔序列促进果蝇核糖体DNA的体外转录。
Nucleic Acids Res. 1982 Nov 11;10(21):6879-86. doi: 10.1093/nar/10.21.6879.
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A protein binds to a satellite DNA repeat at three specific sites that would be brought into mutual proximity by DNA folding in the nucleosome.一种蛋白质在三个特定位点与卫星DNA重复序列结合，这些位点在核小体中会通过DNA折叠而相互靠近。
Cell. 1984 Jul;37(3):889-901. doi: 10.1016/0092-8674(84)90424-0.
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Chromatin structure of hsp 70 genes, activated by heat shock: selective removal of histones from the coding region and their absence from the 5' region.热休克激活的hsp 70基因的染色质结构：组蛋白从编码区的选择性去除及其在5'区域的缺失。
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Genomic sequencing.基因组测序
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