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. 2019 Dec 5;10:5542. doi: 10.1038/s41467-019-13525-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Mass spectrometric analyses of acp³U20 in tRNA^Cys(GCA) isolated from WT (left), DTWD1 KO (left middle), DTWD2 KO (right middle), and DTWD1/DTWD2 double-KO (right) strains. XICs show corresponding negative ions of the RNase T₁-digested fragments as indicated on the right-hand side of each chromatogram. Mass spectra of acp³D- or D-containing fragments overlap with the isotopic peaks of acp³U- or U-containing fragments. The peak intensities of these fragments are normalized in consideration of those isotopes. Asterisk indicates nonspecific peaks with the same m/z value. b Mass spectrometric analyses of acp³U20a in tRNA^Ile(AAT) isolated from WT (left), DTWD1 KO (left middle), DTWD2L KO (middle), DTWD2 KO (right middle), and DTWD1/DTWD2 double-KO (right) strains. c Substrate specificity of human DTWD1 and DTWD2. DTWD1 is responsible for acp³U20 formation in tRNAs for Cys(GCA) and Tyr, whereas DTWD2 is responsible for acp³U20a formation in tRNAs for Asn(GTT), Ile(AAT), and Ile(TAT). DTWD2 also has a weak activity to form acp³U20. The sequence of tRNA^Cys and tRNA^Asn are shown.