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dc.contributor.author
Duss, Olivier
dc.contributor.author
Diarra Dit Konté, Nana
dc.contributor.author
Allain, Frédéric H.-T.
dc.contributor.editor
Kelman, Zvi
dc.date.accessioned
2022-05-05T06:54:02Z
dc.date.available
2017-06-11T21:10:15Z
dc.date.available
2022-05-05T06:54:02Z
dc.date.issued
2015
dc.identifier.isbn
978-0-12-803048-6
en_US
dc.identifier.issn
0076-6879
dc.identifier.other
10.1016/bs.mie.2015.05.029
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/106880
dc.description.abstract
RNA is a crucial regulator involved in most molecular processes of life. Understanding its function at the molecular level requires high-resolution structural information. However, the dynamic nature of RNA complicates structure determination because crystallization is often not possible or can result in crystal-packing artifacts resulting in nonnative structures. To study RNA and its complexes in solution, we described an approach in which large multi-domain RNA or protein–RNA complex structures can be determined at high resolution from isolated domains determined by nuclear magnetic resonance (NMR) spectroscopy, and then constructing the entire macromolecular structure using electron paramagnetic resonance (EPR) long-range distance constraints. Every step in this structure determination approach requires different types of isotope or spin-labeled RNAs. Here, we present a simple modular RNA cut and paste approach including protocols to generate (1) small isotopically labeled RNAs (< 10 nucleotides) for NMR structural studies, which cannot be obtained by standard protocols, (2) large segmentally isotope and/or spin-labeled RNAs for diamagnetic NMR and paramagnetic relaxation enhancement NMR, and (3) large spin-labeled RNAs for pulse EPR spectroscopy.
en_US
dc.language.iso
en
en_US
dc.publisher
Academic Press
en_US
dc.subject
Multiple segmental isotope labeling of RNA
en_US
dc.subject
Nucleic magnetic resonance
en_US
dc.subject
Electron paramagnetic resonance
en_US
dc.subject
Paramagnetic relaxation enhancement
en_US
dc.subject
RNA solution structure determination
en_US
dc.subject
RNA dynamics
en_US
dc.subject
Protein–RNA interactions
en_US
dc.subject
Spin-labeling of RNA
en_US
dc.subject
Small isotope-labeled RNAs for NMR
en_US
dc.subject
RNA ligation
en_US
dc.subject
Site-directed spin-labeling of RNA
en_US
dc.title
Cut and Paste RNA for Nuclear Magnetic Resonance, Paramagnetic Resonance Enhancement, and Electron Paramagnetic Resonance Structural Studies
en_US
dc.type
Book Chapter
ethz.book.title
Isotope Labeling of Biomolecules - Labeling Methods
en_US
ethz.journal.title
Methods in Enzymology
ethz.journal.volume
565
en_US
ethz.pages.start
537
en_US
ethz.pages.end
562
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
New York, NY
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02030 - Dep. Biologie / Dep. of Biology::02517 - Institut für Biochemie / Institute of Biochemistry (IBC)::03591 - Allain, Frédéric / Allain, Frédéric
ethz.date.deposited
2017-06-11T21:10:47Z
ethz.source
ECIT
ethz.identifier.importid
imp593653aff2f0a80545
ethz.ecitpid
pub:167296
ethz.eth
yes
en_US
ethz.availability
Metadata only
en_US
ethz.rosetta.installDate
2017-07-18T12:29:32Z
ethz.rosetta.lastUpdated
2022-03-28T14:16:39Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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