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. 2020 Jul 1;48(14):7899–7913. doi: 10.1093/nar/gkaa560

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© The Author(s) 2020. 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 Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Chemical modifications of uridines in D. discoideum Ax2 wild type and mutant strains of the Elongator-dependent modification pathway. LC–UV–MS/MS analysis of Elongator-dependent modifications of uridines in purified D. discoideum tRNAs. (A) Chemical modifications (blue) of the Elongator-dependent pathway. Shown are the relative abundance of ncm⁵U (no chemical standard available, top) and the absolute abundance of tRNAs carrying the mcm⁵U (center) and mcm⁵s²U (bottom) modifications in the Ax2 wild type and the indicated mutant strains of the Elongator-dependent pathway. The lower limit of quantification (LOQ) is indicated with a dashed line where appropriate. (B) Additional chemical modifications of uridines observed in mutant strains of the Elongator pathway. Shown are the absolute abundance of tRNAs carrying s²U (top), cm⁵U (center) and ncm⁵s²U (bottom) modifications in the Ax2 wild type and the indicated mutant strains of the Elongator-dependent pathway. The LOQ for these modifications is indicated. These modifications (orange) are not observed in the Ax2 wild type and their structures are shown to the right. Please note the different scaling of the y-axes in panels (A) and (B).