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. 2015 May 9;43(11):5617–5629. doi: 10.1093/nar/gkv471

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — The discriminator base affects CCA addition. From 5 min to 4 h of incubation with human CCA-adding enzyme, the incorporation of CCA termini in the human mitochondrial tRNA^Tyr was monitored by the appearance of shifted bands in the polyacrylamide gels. The wild-type transcript with A73 shows the fastest addition, with a complete turnover after 1 h. Transcripts with G73 or U73 show a complete CCA addition after 2 or 4 h, respectively. While the reaction stops after addition of three residues to these substrates, the tRNA ending with C73 shows further band shifts, indicating the incorporation of additional nucleotides.