Conformational flexibility in RNA: The role of dihydrouridine

Joseph J. Dalluge, Takeshi Hashizume, Alan E. Sopchik, James A. McCloskey, Darrell R. Davis

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Abstract

In order to further understand the structural role of the modified nucleoside dihydrouridine in RNA the solution conformations of Dp and ApDpA were analyzed by one- and two-dimensional proton NMR spectroscopy and compared with those of the related uridine-containing compounds. The analyses indicate that dihydrouridine significantly destabilizes the C3'-endo sugar conformation associated with base stacked, ordered, A-type helical RNA. Equilibrium constants (K(eq) = [C2'endo]/[C3'-endo]) for C2'-endo-C3'-endo interconversion at 25°C for Dp, the 5'-terminal A of ApDpA and D in ApDpA are 2.08, 1.35 and 10.8 respectively. Stabilization of the C2'-endo form was shown to be enhanced at low temperature, indicating that C2'-endo is the thermodynamically favored conformation for dihydrouridine. ΔH values show that for Dp the C2'-endo sugar conformation is stabilized by 1.5 kcal/mol compared with Up. This effect is amplified for D in the oligonucleotide ApDpA and propagated to the 5'-neighboring A, with stabilization of the C2'-endo form by 5.3 kcal/mol for D and 3.6 kcal/mol for the 5'-terminal A. Post-transcriptional formation of dihydrouridine therefore represents a biological strategy opposite in effect to ribose methylation, 2-thiolation or pseudouridylation, all of which enhance regional stability through stabilization of the C3'-endo conformer. Dihydrouridine effectively promotes the C2'-endo sugar conformation, allowing for greater conformational flexibility and dynamic motion in regions of RNA where tertiary interactions and loop formation must be simultaneously accomodated.

Original languageEnglish (US)
Pages (from-to)1073-1079
Number of pages7
JournalNucleic acids research
Volume24
Issue number6
DOIs
StatePublished - 1996
Externally publishedYes

Bibliographical note

Funding Information:
The authors wish to thank Jay Olsen for his technical assistance with all NMR experiments and Richard H. Griffey (ISIS Pharmaceuticals) for providing his PSEUROT C program. This work was supported by NIH grant GM29812 to J.A.M. and NSF grant MCB-9317196 to D.R.D. The NMR facility is supported by NIH grants P30 CA42014 and S10 RR06262.

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