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对经紫外线照射的大肠杆菌50S核糖体亚基所诱导的23S RNA二级和三级结构中一系列RNA内交联进行定位。

Localisation of a series of intra-RNA cross-links in the secondary and tertiary structure of 23S RNA, induced by ultraviolet irradiation of Escherichia coli 50S ribosomal subunits.

作者信息

Stiege W, Glotz C, Brimacombe R

出版信息

Nucleic Acids Res. 1983 Mar 25;11(6):1687-706. doi: 10.1093/nar/11.6.1687.

DOI:10.1093/nar/11.6.1687
PMID:6340065
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC325829/
Abstract

Intra-RNA cross-links were introduced into E. coli 50S ribosomal subunits by mild ultraviolet irradiation. The subunits were partially digested with cobra venom nuclease, and the cross-linked RNA complexes were isolated by two-dimensional electrophoresis. Many of the complexes were submitted to a second partial digestion procedure. Oligonucleotide analysis of the RNA fragments obtained in this manner enabled cross-links between the following ribonuclease T1 oligonucleotides in the 23S RNA to be established: positions 292-296 and 339-350; 601-604 and 652-656; 1018-1022 and 1140-1149; 1433-1435 and 1556-1560; 1836-1839 and 1898-1903; 2832-2834 (tentative) and 2878-2885; 2849-2852 and 2865-2867 (tentative); 739-748 and 2609-2618; 571-577 and 2030-2032; 1777-1792 (tentative) and 2584-2588. The first seven of these cross-links lie within the secondary structure of the 23S RNA, whereas the last three are tertiary structural cross-links. The degree of precision of the individual determinations was variable, depending on the nucleotide sequence in the vicinity of the cross-link site concerned.

摘要

通过温和的紫外线照射将RNA内交联引入大肠杆菌50S核糖体亚基。用眼镜蛇毒核酸酶对亚基进行部分消化，通过二维电泳分离交联的RNA复合物。许多复合物进行了第二次部分消化程序。对以这种方式获得的RNA片段进行寡核苷酸分析，从而确定了23S RNA中以下核糖核酸酶T1寡核苷酸之间的交联：位置292 - 296和339 - 350；601 - 604和652 - 656；1018 - 1022和1140 - 1149；1433 - 1435和1556 - 1560；1836 - 1839和1898 - 1903；2832 - 2834（暂定）和2878 - 2885；2849 - 2852和2865 - 2867（暂定）；739 - 748和2609 - 2618；571 - 577和2030 - 2032；1777 - 1792（暂定）和2584 - 2588。这些交联中的前七个位于23S RNA的二级结构内，而最后三个是三级结构交联。各个测定的精确程度各不相同，这取决于相关交联位点附近的核苷酸序列。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/b58ea7956cb8/nar00351-0068-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/7e8244cc15de/nar00351-0065-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/1bb76dc37f50/nar00351-0066-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/b58ea7956cb8/nar00351-0068-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/7e8244cc15de/nar00351-0065-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/1bb76dc37f50/nar00351-0066-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/48fe/325829/b58ea7956cb8/nar00351-0068-a.jpg

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本文引用的文献

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Secondary structure of the large subunit ribosomal RNA from Escherichia coli, Zea mays chloroplast, and human and mouse mitochondrial ribosomes.
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Structural characterization of U*-1915 in domain IV from Escherichia coli 23S ribosomal RNA as 3-methylpseudouridine.来自大肠杆菌23S核糖体RNA结构域IV的U*-1915被鉴定为3-甲基假尿苷的结构特征。
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