NMR Spectroscopy of RNA Duplexes Containing Pseudouridine in Supercooled Water

Overview

Journal RNA

Specialty Molecular Biology

Date 2005 Jul 1

PMID 15987812

Citations 12

Authors

Kersten T Schroeder

Jack J Skalicky

Nancy L Greenbaum

Affiliations

Soon will be listed here.

Abstract

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 degrees 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.

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