15N-labeled tRNA. Identification of 4-thiouridine in Escherichia coli tRNASer1 and tRNATyr2 by 1H-15N two-dimensional NMR spectroscopy.

Publication Type Journal Article
School or College College of Pharmacy; College of Science; School of Medicine
Department Biochemistry; Medicinal Chemistry; Chemistry
Creator Poulter, Charles Dale; Davis, Darrell R.
Other Author Griffey, Richard H.; Yamaizumi, Ziro; Nishimura, Susumu; Hawkins, Bruce
Title 15N-labeled tRNA. Identification of 4-thiouridine in Escherichia coli tRNASer1 and tRNATyr2 by 1H-15N two-dimensional NMR spectroscopy.
Date 1986-09-15
Description Uridine is uniquely conserved at position 8 in elongator tRNAs and binds to A14 to form a reversed Hoogsteen base pair which folds the dihydrouridine loop back into the core of the L-shaped molecule. On the basis of 1H NMR studies, Hurd and co-workers (Hurd, R. E., Robillard, G. T., and Reid, B. R. (1977) Biochemistry 16, 2095-2100) concluded that the interaction between positions 8 and 14 is absent in Escherichia coli tRNAs with only 3 base pairs in the dihydrouridine stem. We have taken advantage of the unique 15N chemical shift of N3 in thiouridine to identify 1H and 15N resonances for the imino units of S4U8 and s4U9 in E. coli tRNASer1 and tRNATyr2. Model studies with chloroform-soluble derivatives of uridine and 4-thiouridine show that the chemical shifts of the protons in the imino moieties move downfield from 7.9 to 14.4 ppm and from 9.1 to 15.7 ppm, respectively; whereas, the corresponding 15N chemical shifts move downfield from 157.5 to 162.5 ppm and from 175.5 to 180.1 ppm upon hydrogen bonding to 5'-O-acetyl-2',3'-isopropylidene adenosine. The large difference in 15N chemical shifts for U and s4U allows one to unambiguously identify s4U imino resonances by 15N NMR spectroscopy. E. coli tRNASer1 and tRNATyr2 were selectively enriched with 15N at N3 of all uridines and modified uridines. Two-dimensional 1H-15N chemical shift correlation NMR spectroscopy revealed that both tRNAs have resonances with 1H and 15N chemical shifts characteristic of s4UA pairs. The 1H shift is approximately 1 ppm upfield from the typical s4U8 resonance at 14.8 ppm, presumably as a result of local diamagnetic anisotropies. An additional s4U resonance with 1H and 15N shifts typical of interaction of a bound water or a sugar hydroxyl group with s4U9 was discovered in the spectrum of tRNATyr2. Our NMR results for tRNAs with 3-base pair dihydrouridine stems suggest that these molecules have an U8A14 tertiary interaction similar to that found in tRNAs with 4-base pair dihydrouridine stems.
Type Text
Publisher American Society for Biochemistry and Molecular Biology (ASBMB)
Volume 261
Issue 26
First Page 12074
Last Page 12078
Subject Magnetic resonance spectroscopy
Subject MESH RNA, Transfer, Amino Acyl; Nucleic Acid Conformation
Language eng
Bibliographic Citation J Biol Chem. 1986 Sep 15;261(26):12074-8. Griffey RH, Davis DR, Yamaizumi Z, Nishimura S, Hawkins BL, Poulter CD. 15N-labeled tRNA. Identification of 4-thiouridine in Escherichia coli tRNASer1 and tRNATyr2 by 1H-15N two-dimensional NMR spectroscopy. Retrieved April 16, 2007 from http://www.jbc.org/cgi/reprint/261/26/12074.
Rights Management Copyright © 1986 American Society for Biochemistry and Molecular Biology. All rights reserved.
Format Medium application/pdf (portable document)
Identifier ir-main,935
ARK ark:/87278/s63r1bfc
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2012-07-11
ID 706282
Reference URL https://collections.lib.utah.edu/details?id=706282
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