Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system

Mesters, J.R.; Vorstenbosch, Erik L.H.; Boer, A.Just De; Kraal, Barend

doi:10.1016/0021-9673(94)80314-5

Cited by 6 publications

(2 citation statements)

References 10 publications

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“…Isolation of tRNA Phe from yeast. Partially purified tRNA Phe was obtained from a crude tRNA mixture from brewer's yeast as previously described (13).…”

Section: Methodsmentioning

confidence: 99%

The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase

Deiman¹,

Kortlever²,

Pleij³

1997

J Virol

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The tRNA-like structure at the 3 end of turnip yellow mosaic virus (TYMV) RNA was studied in order to determine the role of this structure in the initiation of minus-strand synthesis in vitro. Deletions in the 5-to-3 direction up to the pseudoknot structure did not result in a decrease of transcription efficiency. However, transcription efficiency was reduced twofold when a fragment of 21 nucleotides, comprising the 3-terminal hairpin, was used as a template. tRNA Phe from yeast, Escherichia coli 5S rRNA, and the 3-terminal 208 nucleotides of alfalfa mosaic virus RNA 3 could not be transcribed by the RNA-dependent RNA polymerase (RdRp) of TYMV. Various mutations in the sequences of loop regions L1 and L2 or of stem region S1 of the pseudoknot were tested to further investigate the importance of the pseudoknot structure. The results were compared with those obtained in an earlier study on aminoacylation with the same mutants (R. M. W. Mans,

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“…Isolation of tRNA Phe from yeast. Partially purified tRNA Phe was obtained from a crude tRNA mixture from brewer's yeast as previously described (13).…”

Section: Methodsmentioning

confidence: 99%

The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase

Deiman¹,

Kortlever²,

Pleij³

1997

J Virol

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“…Purification and aminoacylation of tRNAs S. cerevisiae tRNA Phe ('chemical block') was aminoacylated according to the protocol as described 31 with minor modifications. After three rounds of phenol-chloroform-isoamyl alcohol extraction S. cerevisiae Phe-tRNA Phe was purified on a column DeltaPack, C4 300A, 5mkm, 3.9 × 150 mm HPLC column (Waters) using a ethanol gradient as described 32 . The final sample was stored in 20 mM NH 4 CH 3 CO 2 pH 5.0 at −80 °C.…”

Section: Purification Of Native Eef2mentioning

confidence: 99%

Accuracy mechanism of eukaryotic ribosome translocation

Djumagulov

Demeshkina

Jenner

et al. 2021

Nature

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Translation of the genetic code into proteins is realized through repetitions of synchronous translocation of messenger RNA (mRNA) and transfer RNAs (tRNA) through the ribosome. In eukaryotes translocation is ensured by elongation factor 2 (eEF2), which catalyses the process and actively contributes to its accuracy1. Although numerous studies point to critical roles for both the conserved eukaryotic posttranslational modification diphthamide in eEF2 and tRNA modifications in supporting the accuracy of translocation, detailed molecular mechanisms describing their specific functions are poorly understood. Here we report a high-resolution X-ray structure of the eukaryotic 80S ribosome in a translocation-intermediate state containing mRNA, naturally modified eEF2 and tRNAs. The crystal structure reveals a network of stabilization of codon–anticodon interactions involving diphthamide1 and the hypermodified nucleoside wybutosine at position 37 of phenylalanine tRNA, which is also known to enhance translation accuracy2. The model demonstrates how the decoding centre releases a codon–anticodon duplex, allowing its movement on the ribosome, and emphasizes the function of eEF2 as a ‘pawl’ defining the directionality of translocation3. This model suggests how eukaryote-specific elements of the 80S ribosome, eEF2 and tRNAs undergo large-scale molecular reorganizations to ensure maintenance of the mRNA reading frame during the complex process of translocation.

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In vitro selection of RNAs aminoacylated by Escherichia coli leucyl-tRNA synthetase

Asahara

Nameki

Hasegawa

1998

Journal of Molecular Biology

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Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system

Cited by 6 publications

References 10 publications

The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase

The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase

Accuracy mechanism of eukaryotic ribosome translocation

In vitro selection of RNAs aminoacylated by Escherichia coli leucyl-tRNA synthetase

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