Reactions at the termini of tRNA with T4 RNA ligase

A. Gregory Bruce; Olke C. Uhlenbeck

doi:10.1093/nar/5.10.3665

Reactions at the termini of tRNA with T4 RNA ligase

A. Gregory Bruce^*, Olke C. Uhlenbeck

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

273 Scopus citations

Abstract

T4 RNA ligase will catalyze the addition of nucleoside 3′,5′-bisphos-phates Onto the 3′ terminus of tRNA resulting in a tRNA molecule one nucleotide longer with a 3′ terminal phosphate. Under appropriate conditions the reaction is quantitative and, if high specific radioactivity bisphosphates are used, it provides an efficient means for in vitro labeling of tRNA. Although the 3′ terminal hydroxyl is a good acceptor, the 5′ terminal phosphate in most tRNA is not an effective donor in the RNA ligase reaction. This poor reactivity is due to the secondary structure of the 5′ terminal nucleotide. If E. coli tRNA_f^Met is used, the 5′ phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.

Original language	English (US)
Pages (from-to)	3665-3678
Number of pages	14
Journal	Nucleic acids research
Volume	5
Issue number	10
DOIs	https://doi.org/10.1093/nar/5.10.3665
State	Published - Oct 1978

Funding

ACKNOWLEDGMENT S We would like to thank P. Cole for the gift of tRNA^", and D. Ward and M. Rabin for discussions on mercuration and iodination. This work was supported by a grant from the National Institutes of Health (GM 19059).

ASJC Scopus subject areas

Genetics

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Cite this

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abstract = "T4 RNA ligase will catalyze the addition of nucleoside 3′,5′-bisphos-phates Onto the 3′ terminus of tRNA resulting in a tRNA molecule one nucleotide longer with a 3′ terminal phosphate. Under appropriate conditions the reaction is quantitative and, if high specific radioactivity bisphosphates are used, it provides an efficient means for in vitro labeling of tRNA. Although the 3′ terminal hydroxyl is a good acceptor, the 5′ terminal phosphate in most tRNA is not an effective donor in the RNA ligase reaction. This poor reactivity is due to the secondary structure of the 5′ terminal nucleotide. If E. coli tRNAfMet is used, the 5′ phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.",

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AB - T4 RNA ligase will catalyze the addition of nucleoside 3′,5′-bisphos-phates Onto the 3′ terminus of tRNA resulting in a tRNA molecule one nucleotide longer with a 3′ terminal phosphate. Under appropriate conditions the reaction is quantitative and, if high specific radioactivity bisphosphates are used, it provides an efficient means for in vitro labeling of tRNA. Although the 3′ terminal hydroxyl is a good acceptor, the 5′ terminal phosphate in most tRNA is not an effective donor in the RNA ligase reaction. This poor reactivity is due to the secondary structure of the 5′ terminal nucleotide. If E. coli tRNAfMet is used, the 5′ phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.

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Reactions at the termini of tRNA with T4 RNA ligase

Abstract

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