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. 2015 May;21(5):935–945. doi: 10.1261/rna.049080.114

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© 2015 Wolf and Grayhack; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 2. — Frameshifting depends upon inefficient decoding of CGA codons and yields a full-length polypeptide. Expression of the nonnative anticodon mutated tRNA^Arg(UCG) (A) or the native tRNA^Arg(ICG) (B) on a 2μ plasmid suppresses frameshifting in the (CGA)₄+1 construct in an asc1-Δ mutant. Median GFP/RFP expression of the indicated R Luc-(NGA)₄-GFP constructs in the indicated wild-type and asc1-Δ strains. (C) CGA-mediated frameshifting yields a full-length fusion protein. Yeast extracts containing indicated units of GFP/RFP were resolved by SDS-PAGE and probed with an anti-HA antibody to detect GFP; the HA epitope is encoded just downstream from the frameshift site in-frame with GFP. The sample in lane k is derived from a GFP construct lacking an initiating methionine, while samples in lanes l–o are derived from yeast extracts expressing a 3C-HA-His6-superfolder GFP protein (lacking Renilla luciferase), derived from pEKD1024 (Dean and Grayhack 2012). Relative amounts of input protein were varied in lanes l–o and lanes f, i, and j, as indicated.