Open AccessNMR spectroscopy of RNA duplexes containing pseudouridine in supercooled water
Author(s) -
Kersten T. Schroeder,
Jack J. Skalicky,
Nancy L. Greenbaum
Publication year - 2005
Publication title -
rna
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.037
H-Index - 171
eISSN - 1469-9001
pISSN - 1355-8382
DOI - 10.1261/rna.2270205
Subject(s) - pseudouridine , rna , duplex (building) , nuclear magnetic resonance spectroscopy , base pair , hydrogen bond , supercooling , crystallography , proton nmr , helix (gastropod) , two dimensional nuclear magnetic resonance spectroscopy , biology , oligonucleotide , stereochemistry , dna , chemistry , molecule , biochemistry , transfer rna , gene , physics , thermodynamics , organic chemistry , ecology , snail
We have performed NMR experiments in supercooled water in order to decrease the temperature-dependent exchange of protons in RNA duplexes. NMR spectra of aqueous samples of RNA in bundles of narrow capillaries that were acquired at temperatures as low as −18°C reveal resonances of exchangeable protons not seen at higher temperatures. In particular, we detected the imino protons of terminal base pairs and the imino proton of a non-base-paired pseudouridine in a duplex representing the eukaryotic pre-mRNA branch site helix. Analysis of the temperature dependence of chemical shift changes (thermal coefficients) for imino protons corroborated hydrogen bonding patterns observed in the NMR-derived structural model of the branch site helix. The ability to observe non-base-paired imino protons of RNA is of significant value in structure determination of RNA motifs containing loop and bulge regions.
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom