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. 2015 Feb 4;43(4):2242–2258. doi: 10.1093/nar/gkv075

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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-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 3. — Human NAT10 is required for 18S rRNA and tRNA acetylation. (A, B and E, F) Human NAT10 substitutes for the function of yeast Kre33 in 18S rRNA and tRNA acetylation. The gene encoding NAT10 was expressed in kre33Δ cells, 18S rRNA (A, B) and tRNAs (E, F) were purified and analyzed by HPLC. Inset: western blotting with anti-NAT10 antibody established that the human construct is stably expressed in yeast cells. (C, D and G, H) NAT10 is responsible for 18S rRNA and tRNA acetylation in human cells: colon carcinoma cells (HCT116 p53+/+) transfected with an siRNA specific to NAT10 (siRNA#1) or with a non-targeting control (scramble) were incubated 72 h, 18S RNA (C, D) and tRNAs (G, H) were purified and analyzed by HPLC. (I) Human NAT10 complements growth of Δkre33 cells to the same extent as control wild-type yeast Kre33. Drop assay showing serial culture dilutions on synthetic medium incubated for 3 days at 30°C.