Complementary-addressed site-directed spin labeling of long natural RNAs

被引：31

作者：

Babaylova, Elena S. ^{[1
,2
]}

Malygin, Alexey A. ^{[1
,2
]}

Lomzov, Alexander A. ^{[1
,2
]}

Pyshnyi, Dmitrii V. ^{[1
,2
]}

Yulikov, Maxim ^{[3
]}

Jeschke, Gunnar ^{[3
]}

Krumkacheva, Olesya A. ^{[2
,4
]}

Fedin, Matvey V. ^{[2
,4
]}

Karpova, Galina G. ^{[1
,2
]}

Bagryanskaya, Elena G. ^{[2
,5
]}

机构：

[1] Inst Chem Biol & Fundamental Med SB RAS, Novosibirsk 630090, Russia

[2] Novosibirsk State Univ, Novosibirsk 630090, Russia

[3] ETH, Phys Chem Lab, CH-8093 Zurich, Switzerland

[4] Int Tomog Ctr SB RAS, Novosibirsk 630090, Russia

[5] NN Vorozhtsov Novosibirsk Inst Organ Chem SB RAS, Novosibirsk 630090, Russia

来源：

NUCLEIC ACIDS RESEARCH | 2016年 / 44卷 / 16期

基金：

俄罗斯科学基金会;

关键词：

HEPATITIS-C VIRUS; PARAMAGNETIC-RESONANCE SPECTROSCOPY; NANOMETER DISTANCE MEASUREMENTS; COMPREHENSIVE SOFTWARE PACKAGE; INDEPENDENT NITROXIDE PROBE; PULSED EPR SPECTROSCOPY; RIBOSOME ENTRY SITE; NUCLEIC-ACIDS; TRANSLATION INITIATION; PELDOR SPECTROSCOPY;

D O I：

10.1093/nar/gkw516

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Nanoscale distance measurements by pulse dipolar Electron paramagnetic resonance (EPR) spectroscopy allow new insights into the structure and dynamics of complex biopolymers. EPR detection requires site directed spin labeling (SDSL) of biomolecule(s), which remained challenging for long RNAs up-to-date. Here, we demonstrate that novel complementary-addressed SDSL approach allows efficient spin labeling and following structural EPR studies of long RNAs. We succeeded to spin-label Hepatitis C Virus RNA internal ribosome entry site consisting of a parts per thousand 330 nucleotides and having a complicated spatial structure. Application of pulsed double electron-electron resonance provided spin-spin distance distribution, which agrees well with the results of molecular dynamics (MD) calculations. Thus, novel SDSL approach in conjunction with EPR and MD allows structural studies of long natural RNAs with nanometer resolution and can be applied to systems of biological and biomedical significance.

引用

页码：7935 / 7943

页数：9

共 29 条

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Huang, Keyun
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 2023, 239
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