Base analysis of ribopolynucleotides by chemical tritium labeling: A methodological study with model nucleosides and purified tRNA species

Randerath, Erika; Yu, Chuan-Tao; Randerath, Kurt

doi:10.1016/0003-2697(72)90181-9

Cited by 148 publications

(58 citation statements)

References 50 publications

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“…A quantitative calculation revealed that this modified nucleoside occurs in B. subtilis tRNAs in nearly molar amounts (table 1). Other conditions and abbreviations as described in Materials and methods or [7] and 181. This result agrees well with the analysis of ribothymidine in B. subtilis tRNAs reported by Randerath [9].…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Occurrence and biosynthesis of ribothymidine in tRNAs of B. subtilis

Arnold¹,

Schmidt²,

Kersten³

1975

FEBS Letters

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Section: Resultsmentioning

confidence: 99%

“…The nucleosides were determined with the technique of Randerath [7]. After digestion of 0.5 AZ6e units to the nucleoside level, the ribose moieties were oxidized with periodate, followed by reduction with ["HI KBH4 (2 Ci/mmol).…”

Section: North-holland Publishing Company -Amsterdammentioning

confidence: 99%

Occurrence and biosynthesis of ribothymidine in tRNAs of B. subtilis

Arnold¹,

Schmidt²,

Kersten³

1975

FEBS Letters

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“…After drying, plates were examined under ultraviolet illumination to localise nucleoside trialcohols. The ten spots seen on all plates (see Randerath et al [14]) were cut out and eluted in 1 ml of 2 M N H 4 0 H for 3 h at room temperature and the amounts of 3H and 14C in the eluates were measured by scintillation counting using 0.8 ml of each eluate and 6.5 ml of a Triton X-IOO/toluene/ PPOjPOPOP mixture [15]. The composition of the 16-S rRNA sample was then calculated.…”

Section: Determination Of' Methylaled Nucleotides In 1 6 3 Rrnamentioning

confidence: 96%

Properties of Ribosomes and Ribosomal RNAs Synthesized by Escherichia coli, Grown in the Presence of Ethionine

Chelbi-Alix

Expert‐Bezançon

Hayes

et al. 2005

European Journal of Biochemistry

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Analysis of 16-S rRNA synthesized in Escherichia coli D10 (met-) incubated in a medium containing ethionine in place of methionine shows that it lacks most and probably all of the methyl groups present in normal 16-S rRNA but possesses the same 3'-OH, and 5'-phosphate terminal sequences as the latter. 23-S rRNA formed in ethioninetreated cells also contains normal terminal sequences. 5-S rRNAs of normal and ethionine-treated E. coli D10 are identical. These results lead to the conclusion that methylation of ribosomal precursor RNAs is not necessary for their maturation to products with iiornial chain lengths and does not influence the conformation of 16-S rRNA.Production of non-methylated cell components can be achieved by culture of Escherichia coli met-mutants in the presence of ethionine [I -41, since this analogue of methionine replaces the latter in all the reactions of protein synthesis [S-71 and does not significantly perturb the functions of proteins in which it replaces methionine [S, 91 ; however, it is not accepted by procaryotic L-methionine S-adenosyltransferases whose product, S-adenosyl-L-methionine, is used for most cellular methylation reactions. Previous publications in this series have described the results of analyses of the protein complements and physicochemical and functional properties of unmethylated ethionine-containing ribosomal subunits prepared in this way. The present report shows that RNA methylation is not required for the correct maturation of the 5-S, 16-S, and 23-S ribosomal RNAs of E. coli.

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“…The active aldehyde radical at position 4324 in 28S rRNA generated by ricin A-chain action was reduced with sodium borohydride by the method of Randerath et al [15]. The modified ribosomes were reacted with 10 mmol/l sodium borohydride to reduce the aldehyde group into hydroxyl group, and then assayed the protein synthesis activity in a reconstituted system.…”

Section: Reduction Of the Active Aldehyde Group With Sodium Borohydridementioning

confidence: 99%

Partial restoration of inactivated ribosomes with sodium borohydride or amino acids

Li¹,

Jin²,

Liu³

1995

FEBS Letters

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The aldehyde radical of ribose C~ at position 4324 in rat liver 28S rRNA generated by RNA N-glycosidase was either reduced to an hydroxyl group by sodium borohydride or converted into aldimine through a nucleophilic addition of amino acid used as a primary amine. Analysis of the R-fragment of 28S rRNA by polyacrylamide gel electrophoresis showed that the reduction of aldehyde to an hydroxyl group with sodium borohydride was highly specific. The protein synthesis activity of modified ribosomes was partially restored with the removal of the active aldehyde by sodium borohydride or amino acid. Reduction of aldehyde with sodium borohydride restored 43.1% of the protein synthesis activity. Among the twenty natural amino acids tested, tryptophan and hlstidine could restore 57.4% and 42.1% of the ribosome activity when hrome mosaic virus RNA was used as mRNA. We came to the conclusion that the active aldehyde radical at position 4324 of 28S rRNA in modified ribosome may cause the inactivation of the ribosome for protein synthesis.

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Base analysis of ribopolynucleotides by chemical tritium labeling: A methodological study with model nucleosides and purified tRNA species

Cited by 148 publications

References 50 publications

Occurrence and biosynthesis of ribothymidine in tRNAs of B. subtilis

Occurrence and biosynthesis of ribothymidine in tRNAs of B. subtilis

Properties of Ribosomes and Ribosomal RNAs Synthesized by Escherichia coli, Grown in the Presence of Ethionine

Partial restoration of inactivated ribosomes with sodium borohydride or amino acids

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