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. 2013 Dec 11;42(5):3246–3260. doi: 10.1093/nar/gkt1281

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© The Author(s) 2013. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), 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. — Mung bean nuclease protection assay. (A) Graphic representation of Mung bean nuclease protection assay used in the present study for the analysis of specific position of modified m³U nucleoside in the Δyil096c and Δylr063w mutants. RP-HPLC chromatogram of the nucleosides derived from protected RNA fragments. Specific fragments of the 25S rRNA from wild type, Δyil096c and Δylr063w corresponding to both m³U2634 and m³U2843 were isolated. The status of m³U residue in these fragments were then analyzed by RP-HPLC. (B) RP-HPLC chromatogram of the fragment corresponding to m³U2634 isolated from the wild type and (C) from Δyil096c. (D) RP-HPLC chromatogram of the fragment corresponding to m³U2843, isolated from the wild type and (E) from Δylr063w. As evident from the chromatograms, the m³U2634 methylation is absent in Δyil096c, whereas m³U2843 is missing in Δylr063w. This clearly demonstrated that Yil096c and Ylr063w are involved in the methylation of m³U2634 and m³U2843, respectively.