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. 2023 Jul 14;51(16):8820–8835. doi: 10.1093/nar/gkad568

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — The proportion of mRNA-bound versus free 80S ribosomes increases in eIF4G1 containing strains but not in eIF4G2-containing strains strain following oxidative stress conditions. (A) Schematic showing the effects of high magnesium concentrations in polysome buffers on either free or mRNA bound 80S ribosomes. Free 80S subunits dissociate to their 40S and 60S constituent parts whereas mRNA bound 80S subunits are stabilised by their mRNA interaction and remain as 80S monosomes. (B) Representative polyribosome traces are shown in the presence or absence of high magnesium for the wild-type and homogenic strains before or after treatment with 0.4 mM H₂O₂ for 15 minutes. Green diamonds (40S), red ovals (60S) and blue triangles (80S) denote ribosomes and subunits. (C) Quantification of the area of the 40S and 60S peaks (free) and the 80S and polysomes (bound) as a proportion of the total trace, where the free fraction represents ribosomal subunits separated by magnesium and the bound fraction represents ribosomes stabilised by mRNA. Significance is shown using a one-way ANOVA test for the bound fraction; *P < 0.05, ***P < 0.001.