+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Effect of nucleoside modifications on the structure and thermal stability of Escherichia coli valine tRNA

Effect of nucleoside modifications on the structure and thermal stability of Escherichia coli valine tRNA

Biochimie 76(12): 1192-1204

Transfer RNA transcribed in vitro lacks the base modifications found in native tRNA. To understand the effect of base modifications on the structure of tRNA, the downfield region of the 1H NMR spectrum of in vitro transcribed E. coli tRNA-Val in aqueous phosphate buffer in the presence of excess Mg-2+ was investigated. The resonances of all imino protons involved in hydrogen bonds in the helical stem regions and in tertiary interactions were assigned using two-dimensional nuclear Overhauser enhancement spectroscopy (NOESY) and one-dimensional difference nuclear Overhauser effect (NOE) methods. In addition, some aromatic C2 and C8 proton resonances as well as one amino proton resonance were assigned. The chemical shifts of the assigned resonances of unmodified E. coli tRNA-Val were compared with those of the native tRNA molecule under similar solution conditions. The similarity of the NMR data for unmodified and modified tRNA indicates that the in vitro transcribed tRNA has nearly the same solution structure as the native molecule in the presence of excess Mg-2+. The only significant differences were the chemical shifts of resonances corresponding to protons in (or interacting with) bases, indicating the possibility of local structural perturbations. The thermal stability of E. coli modified and unmodified tRNA-Val in the presence of Mg-2+ was also investigated by analyzing the temperature dependence of the imino proton spectra. Several tertiary interactions involving modified nucleosides in native E. coli tRNA-Val are less stable in the absence of base modifications.

Please choose payment method:

(PDF emailed within 1 workday: $29.90)

Accession: 008540473

Download citation: RISBibTeXText

PMID: 7538325

Related references

Nucleoside modifications affect the structure and stability of the anticodon of tRNA(Lys,3). Nucleosides and Nucleotides 18(6-7): 1579-1581, 1999

Nucleoside modifications stabilize Mg-2+ binding in Escherichia coli tRNA-Val: An imino proton NMR investigation. Biochemistry 33(30): 8905-8911, 1994

Effect of modified nucleotides on Escherichia coli tRNAGlu structure and on its aminoacylation by glutamyl-tRNA synthetase: Predominant and distinct roles of the mnm5 and s2 modifications of U34. European Journal of Biochemistry 266(3): 1128-1135, 1999

Possible anticodon sequences of tRNA His , tRNA Asm , and tRNA Asp from Escherichia coli B. Universal presence of nucleoside Q in the first postion of the anticondons of these transfer ribonucleic acids. Biochemistry 11(2): 301-308, 1972

Interactions of Escherichia coli SO-187 tRNA(IVal) with Bacillus stearothermophilus valine-tRNA synthetase studied by 13C-NMR. Biochimica et Biophysica Acta 1008(3): 293-300, 1989

Fluorine-19 nuclear magnetic resonance as a probe of the solution structure of mutants of 5-fluorouracil-substituted Escherichia coli valine tRNA. Journal Of Molecular Biology. 227(4): 1164-1172, 1992

Fidelity in the aminoacylation of tRNA(Val) with hydroxy analogues of valine, leucine, and isoleucine by valyl-tRNA synthetases from Saccharomyces cerevisiae and Escherichia coli. Biochemistry 29(34): 7953-7958, 1990

A comparative study of the thermal stability of phenylalanyl tRNA synthetase from Escherichia coli MRE-600 and Thermus thermophilus HB 8. Biofizika 39(5): 783-787, 1994

Studies on valyl-tRNA synthetase and tRNA from Escherichia coli. II. Interaction between valyl-tRNA synthetase and valine acceptor tRNA. Journal of Molecular Biology 44(1): 17-30, 1969

Recognition of the nucleoside in the first position of the anticodon of isoleucine trna by isoleucyl trna synthetase from escherichia coli. Nucleosides & Nucleotides 11(2-4): 719-730, 1992

Nucleotide sequence of valine tRNA 1 from Escherichia coli B. Biochimica et Biophysica Acta 195(2): 590-592, 1969

NMR studies of Bacillus subtilis tRNA(Trp) hyperexpressed in Escherichia coli. Assignment of imino proton signals and determination of thermal stability. Journal of Biological Chemistry 275(10): 6712-6716, 2000

Nucleotide sequence of oligonucleotides derived from Escherichia coli valine transfer RNA by ribonuclease T1 digestion: comparison of the sequences neighboring 3'- and 5'-terminals and anticodon region of Escherichia coli valine transfer RNA with those of yeast valine transfer RNA. Biochimica et Biophysica Acta 182(2): 590-592, 1969

Stability of Escherichia coli alanyl-tRNA synthetase quaternary structure under increased pressure. Archives of Biochemistry and Biophysics 346(2): 322-323, 1997

Comparative studies on the isoleucyl- and leucyl-tRNA synthetases from Bacillus stearothermophilus and Escherichia coli: thermal stability of the aminoacyladenylate-enzyme complexes. Febs Letters 4(3): 239-242, 1969