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. 2021 Jan;27(1):40–53. doi: 10.1261/rna.077339.120

FIGURE 8.

FIGURE 8.

Domain IV mutants stimulate frameshifting into the −1 reading frame. (A) Schematic showing (no RT) the S2 mRNA containing a slippery sequence at positions 381–388 showing the positions of +1 and −1 out-of-frame stop codons. (−1 RT) A mRNA designed to create read-through of the −1 frame UGA stop codon at position 417, which was replaced by a GGA sense codon. (+1 RT) A +1 read-through mRNA in which the out-of-frame +1 UGA stop codon at position 404 was replaced by a GGA sense codon. Frameshifting events can be assigned to the −1 or +1 reading frames according to whether a frameshift product of increased size appears when translating the −1 RT or +1 RT mRNA. (B) SDS gel showing results of translation through the +1 read-through (+1 RT), −1 read-through (−1 RT), and no-read-through (No RT) mRNAs. Products indicated are (S2), full-length S2 protein; (23 kD) 23 kD carboxy-terminal truncated EF-G product; (−1 RT) read-through product in the −1 frame; (−1 FS) frameshift product in the −1 frame; (−2 FS) product of 2 −1 frameshifting events. Appearance of the 16.6 kDa product with the −1 RT mRNA confirms that the 19.5 kDa band is indeed the product of −1 frameshifting.