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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 8. — The box C/D snoRNA U13 is required for human 18S rRNA acetylation. (A) Diagrams depicting the predicted base-pairing between U13 and the 3′ end of 18S rRNA. The interaction is proposed to occur during subunit biogenesis, with substrate residue 1842 bulging out upon U13 binding. (B) U13 is required for efficient rRNA acetylation but not for tRNA acetylation. Top panel: 18S rRNA purified from HCT116 cells depleted of U13 for 72 h was analyzed by HPLC, revealing a 2-fold reduction in acetylation (in red). Control HPLC trace from unperturbed cells in black; middle panel, same analysis for purified tRNAs showing no reduction in acetylation; bottom panel, northern blot analysis of residual level of U13 (normalized with respect to the scramble control). (C) U13 is not required for DIMT1L-mediated 18S rRNA 3′ end dimethylation. Total RNA extracted from cells depleted of U13 or DIMT1L for 72 h was analyzed by primer extension to detect the modification. The percentage of residual dimethylation was normalized with respect to a structural stop (*). The efficiencies of DIMT1L depletion (tested by qRT-PCR) and U13 depletion (tested by quantitative northern blot) are shown underneath.

Inline graphic — The box C/D snoRNA U13 is required for human 18S rRNA acetylation. (A) Diagrams depicting the predicted base-pairing between U13 and the 3′ end of 18S rRNA. The interaction is proposed to occur during subunit biogenesis, with substrate residue 1842 bulging out upon U13 binding. (B) U13 is required for efficient rRNA acetylation but not for tRNA acetylation. Top panel: 18S rRNA purified from HCT116 cells depleted of U13 for 72 h was analyzed by HPLC, revealing a 2-fold reduction in acetylation (in red). Control HPLC trace from unperturbed cells in black; middle panel, same analysis for purified tRNAs showing no reduction in acetylation; bottom panel, northern blot analysis of residual level of U13 (normalized with respect to the scramble control). (C) U13 is not required for DIMT1L-mediated 18S rRNA 3′ end dimethylation. Total RNA extracted from cells depleted of U13 or DIMT1L for 72 h was analyzed by primer extension to detect the modification. The percentage of residual dimethylation was normalized with respect to a structural stop (*). The efficiencies of DIMT1L depletion (tested by qRT-PCR) and U13 depletion (tested by quantitative northern blot) are shown underneath.