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大肠杆菌16S核糖体RNA 3'末端区域的拓扑结构；一项RNA内交联研究。

The topography of the 3'-terminal region of Escherichia coli 16S ribosomal RNA; an intra-RNA cross-linking study.

作者信息

Döring T, Greuer B, Brimacombe R

机构信息

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

出版信息

Nucleic Acids Res. 1992 Apr 11;20(7):1593-7. doi: 10.1093/nar/20.7.1593.

DOI:10.1093/nar/20.7.1593
PMID:1374556
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC312243/
Abstract

30S ribosomal subunits, 70S ribosomes or polysomes from E. coli were subjected to mild ultraviolet irradiation, and the 3'-terminal region of the 16S RNA was excised by 'addressed cleavage' using ribonuclease H in the presence of suitable complementary oligodeoxynucleotides. RNA fragments from this region containing intra-RNA cross-links were separated by two-dimensional gel electrophoresis and the cross-link sites identified by our standard procedures. Five new cross-links were found in the 30S subunit, which were localized at positions 1393-1401 linked to 1531-1532, 1393-1401 linked to 1506, 1393-1401 to 1502-1504, 1402-1403 to 1498-1501, and 1432 to 1465-69, respectively. In 70S ribosomes or polysomes the first four of these were absent, but instead two cross-links between the 1400-region and tRNA were observed. These results are discussed in the context of the tertiary folding of the 3'-terminal region of the 16S RNA and its known functional significance as part of the ribosomal decoding centre.

摘要

对来自大肠杆菌的30S核糖体亚基、70S核糖体或多核糖体进行轻度紫外线照射，然后在合适的互补寡聚脱氧核苷酸存在的情况下，使用核糖核酸酶H通过“定向切割”切除16S RNA的3'末端区域。通过二维凝胶电泳分离该区域中含有RNA内交联的RNA片段，并通过我们的标准程序确定交联位点。在30S亚基中发现了五个新的交联，它们分别位于1393 - 1401与1531 - 1532相连、1393 - 1401与1506相连、1393 - 1401与1502 - 1504相连、1402 - 1403与1498 - 1501相连以及1432与1465 - 69相连的位置。在70S核糖体或多核糖体中，前四个交联不存在，但观察到1400区域与tRNA之间有两个交联。这些结果将在16S RNA 3'末端区域的三级折叠及其作为核糖体解码中心一部分的已知功能意义的背景下进行讨论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6845/312243/11a8cf4f7793/nar00081-0148-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6845/312243/11a8cf4f7793/nar00081-0148-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6845/312243/11a8cf4f7793/nar00081-0148-a.jpg

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本文引用的文献

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Covalent crosslinking of tRNA1Val to 16S RNA at the ribosomal P site: identification of crosslinked residues.tRNA1Val在核糖体P位点与16S RNA的共价交联：交联残基的鉴定。
Proc Natl Acad Sci U S A. 1982 Sep;79(18):5450-4. doi: 10.1073/pnas.79.18.5450.
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The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.大肠杆菌16S核糖体RNA的3'末端序列：与无义三联体及核糖体结合位点的互补性
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Intra-RNA and RNA-protein cross-linking techniques in Escherichia coli ribosomes.
从不断演变的核糖体RNA中汲取的经验教训：从比较的角度看16S和23S核糖体RNA结构
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Probing the conformational changes in 5.8S, 18S and 28S rRNA upon association of derived subunits into complete 80S ribosomes.探究5.8S、18S和28S核糖体RNA（rRNA）在衍生亚基缔合形成完整80S核糖体时的构象变化。
Nucleic Acids Res. 1994 Jul 25;22(14):2776-83. doi: 10.1093/nar/22.14.2776.
5
Three widely separated positions in the 16S RNA lie in or close to the ribosomal decoding region; a site-directed cross-linking study with mRNA analogues.16S核糖体RNA中三个相距甚远的位置位于核糖体解码区域内或其附近；一项使用mRNA类似物的定点交联研究。
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大肠杆菌核糖体中的RNA内部及RNA-蛋白质交联技术
Methods Enzymol. 1988;164:287-309. doi: 10.1016/s0076-6879(88)64050-x.
4
Investigation of the tertiary folding of Escherichia coli 16S RNA by in situ intra-RNA cross-linking within 30S ribosomal subunits.通过30S核糖体亚基内的原位RNA内交联研究大肠杆菌16S RNA的三级折叠。
Nucleic Acids Res. 1985 Oct 11;13(19):6919-36. doi: 10.1093/nar/13.19.6919.
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Interaction of antibiotics with functional sites in 16S ribosomal RNA.抗生素与16S核糖体RNA功能位点的相互作用。
Nature. 1987;327(6121):389-94. doi: 10.1038/327389a0.
6
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Model for the three-dimensional folding of 16 S ribosomal RNA.16S核糖体RNA的三维折叠模型
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