Measurement of imino 1H-1H residual dipolar couplings in RNA

Michael P. Latham; Arthur Pardi

doi:10.1007/s10858-008-9293-8

Measurement of imino ¹H-¹H residual dipolar couplings in RNA

Michael P. Latham, Arthur Pardi

Chemistry and Biochemistry

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Imino ¹H-¹⁵N residual dipolar couplings (RDCs) provide additional structural information that complements standard ¹H-¹H NOEs leading to improvements in both the local and global structure of RNAs. Here, we report measurement of imino ¹H-¹H RDCs for the Iron Responsive Element (IRE) RNA and native E. coli tRNA^Val using a BEST-Jcomp-HMQC2 experiment. ¹H-¹H RDCs are observed between the imino protons in G-U wobble base pairs and between imino protons on neighboring base pairs in both RNAs. These imino ¹H- ¹H RDCs complement standard ¹H-¹⁵N RDCs because the ¹H-¹H vectors generally point along the helical axis, roughly perpendicular to ¹H- ¹⁵N RDCs. The use of longitudinal relaxation enhancement increased the signal-to-noise of the spectra by ∼3.5-fold over the standard experiment. The ability to measure imino ¹H- ¹H RDCs offers a new restraint, which can be used in NMR domain orientation and structural studies of RNAs.

Original language	English
Pages (from-to)	121-129
Number of pages	9
Journal	Journal of Biomolecular NMR
Volume	43
Issue number	2
DOIs	https://doi.org/10.1007/s10858-008-9293-8
State	Published - 2009

Keywords

Iron responsive element RNA
Partial alignment
RNA structure
Residual dipolar couplings
tRNA

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title = "Measurement of imino 1H-1H residual dipolar couplings in RNA",

abstract = "Imino 1H-15N residual dipolar couplings (RDCs) provide additional structural information that complements standard 1H-1H NOEs leading to improvements in both the local and global structure of RNAs. Here, we report measurement of imino 1H-1H RDCs for the Iron Responsive Element (IRE) RNA and native E. coli tRNAVal using a BEST-Jcomp-HMQC2 experiment. 1H-1H RDCs are observed between the imino protons in G-U wobble base pairs and between imino protons on neighboring base pairs in both RNAs. These imino 1H- 1H RDCs complement standard 1H-15N RDCs because the 1H-1H vectors generally point along the helical axis, roughly perpendicular to 1H- 15N RDCs. The use of longitudinal relaxation enhancement increased the signal-to-noise of the spectra by ∼3.5-fold over the standard experiment. The ability to measure imino 1H- 1H RDCs offers a new restraint, which can be used in NMR domain orientation and structural studies of RNAs.",

keywords = "Iron responsive element RNA, Partial alignment, RNA structure, Residual dipolar couplings, tRNA",

author = "Latham, {Michael P.} and Arthur Pardi",

note = "Funding Information: Acknowledgements We thank Dr. J. Boisbouvier for providing the original pulse sequence and Dr. A. Bax for valuable discussions. This work was supported in part by NIH grant AI33098, and MPL was supported in part by a NIH training grant T32 GM65103. The NMR instrumentation was purchased with partial support from NIH grant RR11969 and NSF grants 9602941 and 0230966.",

year = "2009",

doi = "10.1007/s10858-008-9293-8",

language = "English",

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journal = "Journal of Biomolecular NMR",

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AU - Pardi, Arthur

N1 - Funding Information: Acknowledgements We thank Dr. J. Boisbouvier for providing the original pulse sequence and Dr. A. Bax for valuable discussions. This work was supported in part by NIH grant AI33098, and MPL was supported in part by a NIH training grant T32 GM65103. The NMR instrumentation was purchased with partial support from NIH grant RR11969 and NSF grants 9602941 and 0230966.

PY - 2009

Y1 - 2009

N2 - Imino 1H-15N residual dipolar couplings (RDCs) provide additional structural information that complements standard 1H-1H NOEs leading to improvements in both the local and global structure of RNAs. Here, we report measurement of imino 1H-1H RDCs for the Iron Responsive Element (IRE) RNA and native E. coli tRNAVal using a BEST-Jcomp-HMQC2 experiment. 1H-1H RDCs are observed between the imino protons in G-U wobble base pairs and between imino protons on neighboring base pairs in both RNAs. These imino 1H- 1H RDCs complement standard 1H-15N RDCs because the 1H-1H vectors generally point along the helical axis, roughly perpendicular to 1H- 15N RDCs. The use of longitudinal relaxation enhancement increased the signal-to-noise of the spectra by ∼3.5-fold over the standard experiment. The ability to measure imino 1H- 1H RDCs offers a new restraint, which can be used in NMR domain orientation and structural studies of RNAs.

AB - Imino 1H-15N residual dipolar couplings (RDCs) provide additional structural information that complements standard 1H-1H NOEs leading to improvements in both the local and global structure of RNAs. Here, we report measurement of imino 1H-1H RDCs for the Iron Responsive Element (IRE) RNA and native E. coli tRNAVal using a BEST-Jcomp-HMQC2 experiment. 1H-1H RDCs are observed between the imino protons in G-U wobble base pairs and between imino protons on neighboring base pairs in both RNAs. These imino 1H- 1H RDCs complement standard 1H-15N RDCs because the 1H-1H vectors generally point along the helical axis, roughly perpendicular to 1H- 15N RDCs. The use of longitudinal relaxation enhancement increased the signal-to-noise of the spectra by ∼3.5-fold over the standard experiment. The ability to measure imino 1H- 1H RDCs offers a new restraint, which can be used in NMR domain orientation and structural studies of RNAs.

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