Complementary-addressed site-directed spin labeling of long natural RNAs.
Nucleic Acids Res
; 44(16): 7935-43, 2016 09 19.
Article
em En
| MEDLINE
| ID: mdl-27269581
ABSTRACT
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 ≈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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Marcadores de Spin
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RNA Viral
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Hepacivirus
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article