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核糖体RNA的三维折叠；用双（2-氯乙基）甲胺处理大肠杆菌50S核糖体亚基诱导的23S RNA中一系列RNA内交联的定位。

The three-dimensional folding of ribosomal RNA; localization of a series of intra-RNA cross-links in 23S RNA induced by treatment of Escherichia coli 50S ribosomal subunits with bis-(2-chloroethyl)-methylamine.

作者信息

Döring T, Greuer B, Brimacombe R

机构信息

Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, FRG.

出版信息

Nucleic Acids Res. 1991 Jul 11;19(13):3517-24. doi: 10.1093/nar/19.13.3517.

DOI:10.1093/nar/19.13.3517
PMID:1712937
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC328374/
Abstract

Intact 50S ribosomal subunits from E.coli were cross-linked with the symmetrical bifunctional reagent bis-(2-chloroethyl)-methylamine. After deproteinization, selected regions of the 23S RNA were excised by treatment with ribonuclease H in the presence of appropriate complementary decadeoxynucleotides, and screened for the presence of intra-RNA cross-links by two-dimensional gel electrophoresis. Individual isolated cross-linked RNA fragments were analysed by our established procedures. Sixteen intra-RNA cross-links were identified, three of which corresponded to those previously published. The thirteen 'new' cross-links were localized in the 23S RNA at positions 774-78 linked to 792-94, 876-79 linked to 899-900, 979-81 or 983-84 to 2029, 1715 to 1743-46, 1911-21 to 1964, 1933 to 1966, 2032 to 2054-55, 2112 to 2169-71, 2116-17 to 2163-67, 2128-32 to 2156-59, 2392-93 to 2422-23, 2737-38 to 2763-66, and 2791 to 2890. These results are discussed in the context of three-dimensional model-building studies with the 23S RNA, with particular reference to the environment of the 'active centre' of the 50S subunit.

摘要

来自大肠杆菌的完整50S核糖体亚基与对称双功能试剂双（2-氯乙基）甲胺发生交联。脱蛋白后，在适当的互补十脱氧核苷酸存在下，用核糖核酸酶H处理，切除23S RNA的选定区域，并通过二维凝胶电泳筛选RNA内交联的存在。通过我们既定的程序分析各个分离的交联RNA片段。鉴定出16个RNA内交联，其中3个与先前发表的交联相对应。这13个“新”交联位于23S RNA的774 - 78位与792 - 94位相连、876 - 79位与899 - 900位相连、979 - 81或983 - 84位与2029位相连、1715位与1743 - 46位相连、1911 - 21位与1964位相连、1933位与1966位相连、2032位与2054 - 55位相连、2112位与2169 - 71位相连、2116 - 17位与2163 - 67位相连、2128 - 32位与2156 - 59位相连、2392 - 93位与2422 - 23位相连、2737 - 38位与2763 - 66位相连以及2791位与2890位相连。结合对23S RNA的三维模型构建研究结果对这些结果进行了讨论，特别提及了50S亚基“活性中心”的环境。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/0e882e7c96ab/nar00093-0047-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/1a3d62210325/nar00093-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/68d30a995021/nar00093-0047-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/0e882e7c96ab/nar00093-0047-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/1a3d62210325/nar00093-0045-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/68d30a995021/nar00093-0047-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/848d/328374/0e882e7c96ab/nar00093-0047-b.jpg

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