Reactions at the termini of tRNA with T4 RNA ligase

Abstract

T4 RNA ligase will catalyze the addition of nucleoside 3', 5'-bisphosphates onto the 3' terminus of tRNA resulting in 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's 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 tRNAf Met is used, the 5' phosphate is reactive and the major product with RNA ligase is the cyclic tRNA.

Higher-order structure of bovine mitochondrial tRNASerUGA: chemical modification and computer modeling

“…*To whom correspondence should be addressed Chemical probing of tRNAserUGA Bovine mt tRNAserUGA was purified from bovine liver mitochondria as described previously (14), and labeled at the 3'end as described in the literature (15). Modification of the endlabeled tRNA and detection of the modified residues were performed as described previously (9,16).…”

Complementarity Between the 5'- and 3'-terminal Sequences of Rice Stripe Virus RNAs

“…The 3' end-labelling of each RNA species (1 to 3 ~tg) was done as described by Bruce & Uhlenbeck (1978).…”

The nucleotide sequence of sheep liver histidine‐tRNA (anticodon Q‐U‐G)

“…Sequencing of the oligonucleotides and 5'-end labelling of tRNA was performed according to [ 8].3'-End labelling was done as in [9]. The analysis of 2 large fragments isolated from a partial pancreatic digest provided additional results.…”