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. 2017 Feb 6;114(8):E1554–E1563. doi: 10.1073/pnas.1612460114

Fig. 5.

Fig. 5.

Programmable RNA recognition and cleavage by SOT1. (A) The PPR code (9–11). (B) Alignment between Gm-SOT1 and its RNA targets. Individual boxes indicate PPR motifs. The amino acids at positions 5 and 35 of each PPR motif are shown in the box. RNA13 is the minimal binding sequence of SOT1. RNA73 is the sequence of the 5′ region of the 23S–4.5S precursor, including the binding and cleavage sites of SOT1. (C) Alignment between the mutated Gm-SOT1m and its mutated RNA targets. Gm-SOT1m is a mutated Gm-SOT1 protein in which the fifth amino acids in PPR motifs 1–4 (red boxes) were mutated. Mutated ribonucleotides in RNA13m and RNA73m are indicated in red. (D) Programmable RNA recognition by Gm-SOT1. EMSA shows the binding of Gm-SOT1 and the variant Gm-SOT1m to Cy5-labeled RNA13 and Cy3-labeled RNA13m. Increasing concentrations of protein (100, 200, and 400 nM) were incubated with 40 nM probes. (E) Programmable RNA cleavage by Gm-SOT1. RNA cleavage analysis shows the cleavage of FAM-labeled RNA73 and Cy3-labeled RNA73m by Gm-SOT1 and the variant Gm-SOT1m, respectively. Increasing concentrations of protein (100, 200, and 400 nM) were incubated with 40-nM probes.