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. Author manuscript; available in PMC: 2020 Apr 2.
Published in final edited form as: Biochem Biophys Res Commun. 2019 Feb 21;511(2):416–421. doi: 10.1016/j.bbrc.2019.02.073

Fig. 4.

Fig. 4.

Comparison of pseudouridine-modified and unmodified U2 snRNA – BPS-containing duplexes. (a) NOESY cross-peak volumes from the branch site region of the pseudouridine-modified (green) and unmodified (purple) RNA duplexes. The average H1′–H6/H8 and H2′– (n+1)H8/6 NOE volumes of the base-paired stem regions (nucleotides 1–6, 9–10, 33–34, 36–41) are given for comparison with the branch site nucleotides. (b) Correlation of pseudouridine-modified versus unmodified proton shifts (branch site-region nucleotides U6-C9, filled red; other nucleotides 1–5, 10–11, 33, 37–42, unfilled). To expand the region of interest, the shifts of H6/H8/H2, H5/H1´, and H2´/H3´/H4´ protons are expressed relative to 7.5, 5.5, and 4.5 ppm, respectively. The assigned chemical shifts are listed in Table S2. (c) Overlay of natural abundance [13C,1H]-HSQC spectra of pseudouridine-modified (green) and unmodified (purple) duplexes including the C6/C8 (top) and adenine C2 regions (bottom). (d) Proton shift correlations in the presence of 80 mM NaCl and 1.75 mM MgCl2, plotted as in (b). Samples for (a) and (b) were in 100% D2O. Samples for (c) and (d) were in 95% H2O/5% D2O.