Enzymatic replacement of nucleotide sequences in the D-loop of T. utilis tRNATyr and its effect on aminoacylation.
An efficient method for replacement of nucleotide sequences in the D-loop of T. utilis tRNATyr has been developed. An abnormal tRNATyr lacking in tetranucleotide D16-D-Gm-G19 in its D-loop has been reconstructed by this method and shown to accept tyrosine to about 55% of the aminoacylation level observed for intact tRNATyr. This suggests that the deleted sequence itself is not essential for recognition by TyrRS but a conformational instability of the tRNA possibly caused by the disruption of tertiary interactions between the D-loop and T psi C-loop might have influenced the forward reaction rate leading to the decreased level of aminoacylation.
D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism*; D001483:Base Sequence; D002175:Candida; D009690:Nucleic Acid Conformation; D012343:RNA, Transfer / Q000378:metabolism*; D013329:Structure-Activity Relationship; D046249:Transfer RNA Aminoacylation; D014448:Tyrosine-tRNA Ligase / Q000378:metabolism*
D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases; D014448:Tyrosine-tRNA Ligase