Characterization of a heat-stable enzyme possessing GTP-dependent RNA ligase activity from a hyperthermophilic archaeon, Pyrococcus furiosus

Consensus ribonucleotide sequences in tRNA anticodon loop structure are required for ligation reaction by PF0027 protein. Three different substrates bearing 2′,3′-cyclic phosphate and 5′-OH termini were used for the ligation reaction. The reaction was carried out with 0–50 ng of purified PF0027 protein for 15 min at 37°C in the presence of 1 mM GTPγS. (Circles) Mutated ribonucleotides. The substrates used were (A) tRNA5S-P and tRNA3S-FAM, (B) tRNA5SM1-P and tRNA3S-FAM, and (C) tRNA5SM2-P and tRNA3S-FAM.