Enzymatic replacement of nucleotide sequences in the D-loop of T. utilis tRNATyr and its effect on aminoacylation |
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Authors: | T Ohyama K Nishikawa S Takemura |
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Abstract: | 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. |
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