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运用不同荧光技术对含有前体信使核糖核酸的核核糖核蛋白颗粒进行结构研究。

Structural investigation of nuclear RNP particles containing pre-mRNA by different fluorescence techniques.

作者信息

Borissova O F, Krichevskaya A A, Samarina O P

出版信息

Nucleic Acids Res. 1981 Feb 11;9(3):663-81. doi: 10.1093/nar/9.3.663.

DOI:10.1093/nar/9.3.663
PMID:7220348
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC327229/
Abstract

Ethidium bromide (EB) adsorption isotherms on 30S nuclear RNP particles isolated from liver nuclei has revealed 6% of double-stranded regions in pre-mRNA (dsRNA). It has been established by measurements of the EB fluorescence polarization that the bulk of dsRNA regions in RNP is rigidly attached to RNP. They are longer than 45 degree A. The increase of NaCl concentration from 0.1 up to 0.4 M causes a significant loosening of dsRNA-protein bonds. As a result the dsRNA segments become more flexible. Measurements of energy transfer from fluorescamine (covalently bound to the protein) to EB (adsorbed on dsRNA) have yielded information about dsRNA location. The fact that absorbtion of exciting light by fluorescamine causes pronounced increase of EB fluorescence is consistent with the idea that helical regions of RNA are located outside the RNP particles.

摘要

溴化乙锭（EB）在从肝细胞核中分离出的30S核核糖核蛋白（RNP）颗粒上的吸附等温线显示，前体信使核糖核酸（pre-mRNA，双链RNA）中双链区域占6%。通过测量EB荧光偏振确定，核糖核蛋白中大部分双链RNA区域与核糖核蛋白紧密相连。它们的长度超过45埃。氯化钠浓度从0.1M增加到0.4M会导致双链RNA与蛋白质的键显著松弛。结果，双链RNA片段变得更加灵活。对从荧光胺（共价结合到蛋白质上）到EB（吸附在双链RNA上）的能量转移测量得出了有关双链RNA位置的信息。荧光胺吸收激发光导致EB荧光显著增强这一事实与RNA螺旋区域位于核糖核蛋白颗粒外部的观点一致。

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本文引用的文献

1
Polarized fluorescence of acridine orange-transfer RNA complexes.吖啶橙 - 转运RNA复合物的偏振荧光
FEBS Lett. 1970 Jun 8;8(4):201-204. doi: 10.1016/0014-5793(70)80263-0.
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Binding isotherms of tRNA-acriflavine complexes.转运核糖核酸-吖啶黄素复合物的结合等温线
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A fluorescent complex between ethidium bromide and nucleic acids. Physical-chemical characterization.溴化乙锭与核酸之间的荧光复合物。物理化学特性
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Structural organization of nuclear complexes containing DNA-like RNA.含有类DNA RNA的核复合物的结构组织
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Calculation of binding isotherms for heterogenous polymers.非均相聚合物结合等温线的计算。
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Resonance energy transfer between ethidium bromide molecules bound to nucleic acids. Does intercalation wind or unwind the DNA helix?与核酸结合的溴化乙锭分子间的共振能量转移。嵌入作用会使DNA螺旋缠绕还是解旋？
J Mol Biol. 1971 Jul 14;59(1):43-62. doi: 10.1016/0022-2836(71)90412-8.
9
Decay of fluorescence emission anisotropy of the ethidium bromide-DNA complex. Evidence for an internal motion in DNA.溴化乙锭 - DNA复合物荧光发射各向异性的衰减。DNA内部运动的证据。
Proc Natl Acad Sci U S A. 1970 Feb;65(2):417-21. doi: 10.1073/pnas.65.2.417.
10
D-RNA containing ribonucleoprotein particles.含有核糖核蛋白颗粒的D-RNA
Adv Cell Biol. 1971;2:47-110. doi: 10.1007/978-1-4615-9588-5_2.

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