FIGURE 2.

CTSBP2 specifically enhances the reactivity of nucleotides in H89. A, electrophoretic traces obtained from reactions containing 80S ribosomes treated with 80 mm BzCN in the presence (red) or absence (green) of recombinant CT-SBP2, or DMSO (blue). cDNA fragments were generated from primer extension reactions using primer 4168. B, same as in A except primer extension was performed with a second primer (primer 4229) located further downstream (see “Experimental Procedures”). C, Same as A except reactions were performed with the CTSBP2 ⁶⁴⁷RFQDR⁶⁵¹ penta-alanine mutant (purple). D, same as B except reactions were performed with ribosomal protein L30 (rpL30, orange). The y axis shows the relative fluorescence intensity, and the x axis shows the approximate nucleotide position on the mouse 28 S rRNA. Arrows indicate the nucleotides displaying enhanced reactivity in the presence of CTSBP2 (A and B) or the corresponding positions in reactions performed with the CTSBP2 ⁶⁴⁷RFQDR⁶⁵¹ penta-alanine mutant (C) or rpL30 (D). Each trace is representative of at least three independent SHAPE experiments.