An improved method for the separation and quantitation of the modified nucleosides of transfer RNA.

Nucleic acids research ·Vol. 3 ·No. 1 ·1976-01-00 ·Pages 285-95

Rogg H, Brambilla R, Keith G, Staehelin M

Abstract

A method is described which allows a very efficient determination of the modified nucleosides of tRNA. The technique involves enzymatic degradation of the tRNA to nucleosides at pH 7.6 and their separation by two-dimensional thin-layer chromatography on cellulose-coated aluminum foils. Based on the analysis of two mammalian tRNAs it is shown that the technique is suitable for the determination of chemically unstable nucleosides as well as the ribose-methylated compounds. At least 36 of the 45 known modified nucleosides can be separated and quantitatively determined by the method described. This procedure is especially suitable for the estimation of the nucleoside composition of unlabeled tRNAs as well as for studying the post-transcriptional modifications of tRNA.

MeSH Terms

Aluminum Cellulose Chromatography, Thin Layer/methods RNA, Transfer Ribonucleases Ribonucleosides/isolation & purification

Chemicals

Ribonucleosides Cellulose RNA, Transfer Aluminum Ribonucleases

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Rogg H

Brambilla R

Keith G

Staehelin M

References (28)

28 references, click to expand

The effect of growth temperatures on the in vivo ribose methylation of Bacillus stearothermophilus transfer RNA.
Arch Biochem Biophys. 1973 Jan;154(1):277-82 PMID: 4689778
Analysis of methylated constituents from RNA by thin-layer chromatography.
Biochim Biophys Acta. 1967 Apr 18;138(2):430-2 PMID: 6058129
Chromatographic studies of nucleic acids. 4. The nucleic acid of the turnip yellow mosaic virus including a note on the nucleic acid of the tomato bushy stunt virus.
Biochem J. 1951 Sep;49(4):401-6 PMID: 14886301
Ion-exchange thin-layer chromatography. 18. Detection of purine derivatives in the nanogram range by phosphorescence at 77 degrees K.
Anal Biochem. 1967 Dec;21(3):480-5 PMID: 5582136
Ion-exlusion chromatography: analysis and isolation of nucleic acid components, and influence of separation parameters.
Arch Biochem Biophys. 1972 Oct;152(2):800-10 PMID: 4635791
On the bias of the distribution of the 2'-O-methylribonucleotide constituents of yeast transfer RNA.
Biochim Biophys Acta. 1968 Dec 17;169(2):285-96 PMID: 4302867
STRUCTURE OF A RIBONUCLEIC ACID.
Science. 1965 Mar 19;147(3664):1462-5 PMID: 14263761
SOME NUCLEOTIDE SEQUENCES FROM PARTIALLY PURIFIED TRANSFER RIBONUCLEIC ACIDS.
Biochim Biophys Acta. 1964 Jun 22;87:199-211 PMID: 14192361
Methylated bases of transfer ribonucleic acid from HeLa and L cells.
Arch Biochem Biophys. 1968 Mar 20;124(1):472-82 PMID: 5661617
The primary structure of two mammalian tRNAs Phe: identity of calf liver and rabbit liver tRNAs Phe.
FEBS Lett. 1974 Nov 1;48(1):50-2 PMID: 4609801
Oligonucleotide sequences of RNase T 1 and pancreatic RNase digests of E. coli aspartic acid tRNA.
Biochem Biophys Res Commun. 1972 Dec 18;49(6):1605-9 PMID: 4565380
The sequence of phenylalanine tRNA from E. coli.
FEBS Lett. 1969 Jun;3(4):275-278 PMID: 11947028
The separation and estimation of ribonucleotides in minute quantities.
J Biol Chem. 1950 Sep;186(1):37-50 PMID: 14778802
A fast and sensitive method for the analysis of modified nucleosides in tRNA.
Anal Biochem. 1974 Apr;58(2):578-91 PMID: 4363853
Transfer ribonucleic acid from Mycoplasma laidlawii A.
Eur J Biochem. 1971 Feb;18(4):573-81 PMID: 5545013
THIN-LAYER CHROMATOGRAPHY OF NUCLEOTIDES, NUCLEOSIDES AND NUCLEIC ACID BASES.
Biochim Biophys Acta. 1965 Jan 11;95:1-7 PMID: 14289030
Methylation of ribonucleic acid by the carcinogens dimethyl sulphate, N-methyl-N-nitrosourea and N-methyl-N'-nitro-N-nitrosoguanidine. Comparisons of chemical analyses at the nucleoside and base levels.
Biochem J. 1972 Jun;128(1):117-32 PMID: 4673570
The determination of 2'-O-methylnucleosides in RNA.
Anal Biochem. 1974 Oct;61(2):367-78 PMID: 4370836
Nucleotide sequences of rat liver serine-tRNA. 2. The products of digestion with ribonuclease T.
Eur J Biochem. 1971 Jul 29;21(2):243-8 PMID: 5105576
Primary sequence of glutamic acid tRNA II from Escherichia coli.
FEBS Lett. 1972 Feb 1;20(2):239-241 PMID: 11946426
Studies on polynucleotides, lxviii the primary structure of yeast phenylalanine transfer RNA.
Proc Natl Acad Sci U S A. 1967 Mar;57(3):751-8 PMID: 16591527
ON THE SPECIFICITY OF RNAASE T1.
Biochim Biophys Acta. 1964 Jul 22;87:493-5 PMID: 14211643
Base analysis of ribopolynucleotides by chemical tritium labeling: a methodological study with model nucleosides and purified tRNA species.
Anal Biochem. 1972 Jul;48(1):172-98 PMID: 4339367
[Extension of the thin-layer chromatography technique using prefabricated roll preparations].
J Chromatogr. 1970 Apr 8;48(1):144-60 PMID: 5459022
A method for determination of the methylated constituents of transfer ribonucleic acid.
Biochemistry. 1974 Oct 8;13(21):4409-16 PMID: 4369721
Regulation of tRNA.
Annu Rev Biochem. 1973;42:439-70 PMID: 4199854
Determination of the extent of methylation of tRNA.
Anal Biochem. 1975 Apr;64(2):537-44 PMID: 1093443
Derivatives of N-(N-(9-beta-D-ribofuranosylpurin-6-yl)carbamoyl)threonine in phosphodiesterase hydrolysates of wheat embryo transfer ribonucleic acid.
Biochemistry. 1974 Jan 29;13(3):543-53 PMID: 4358950

Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/3.1.285

Published

1976-01-00

Pages

285-95

Language

English

Region

England

NLM ID

0411011

PMCID

PMC342895

Subset

External Links

PubMed source DOI: 10.1093/nar/3.1.285 PMC Full Text View journal details

Analysis Services

An improved method for the separation and quantitation of the modified nucleosides of transfer RNA.

Contact

WeChat Official Account