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• W2896738062 abstract "Abstract The chloroplast, a photosynthetic organelle found in all plant and algae species, originates from an ancient event in which a cyanobacterium was engulfed by a larger eukaryote. Thus, modern chloroplasts still harbor a bacterial-like genome and carry out all stages of gene expression, including mRNA translation by a 70S ribosome. However, the Shine-Dalgarno model, which predominantly regulates translation initiation by base-pairing between the ribosomal RNA and the mRNA in model bacteria genera, was reported to have ambiguous effects on chloroplast gene expression. Here we show that while the Shine-Dalgarno motif is clearly conserved in proteobacterial mRNAs, its general absence from chloroplast mRNAs is observed in cyanobacteria as well, promoting the idea that the evolutionary process of reducing the centrality of the Shine-Dalgarno mechanism began well before plastid endosymbiosis. As plastid ribosomal RNA anti-Shine-Dalgarno elements are highly similar to their bacterial counterparts, these sites alone cannot explain the decline in plastid Shine-Dalgarno generality. However, by computational simulation we show that upstream point mutations modulate the local structure of ribosomal RNA in chloroplasts, creating significantly tighter structures around the anti-Shine-Dalgarno locus, which in-turn reduce the probability of ribosome binding via the Shine-Dalgarno mechanism. To validate our model, we expressed a mCherry gene harboring a Shine-Dalgarno motif in the Chlamydomonas reinhardtii chloroplast. We show that co-expressing it with a 16S ribosomal RNA, modified according to our model, significantly enhances its expression compared to co-expression with an endogenous 16S gene. Significance statement Chloroplasts are fascinating intracellular organelles which have evolved from an ancient cyanobacterium engulfed by a larger eukaryote. Surprisingly, the canonical mechanism regulating bacterial translation initiation – Shine-Dalgarno – has been shown to play a reduced role in chloroplasts. Here, we show that mutations upstream from the anti-Shine-Dalgarno element decrease the probability of spontaneous ribosome binding by modulating the secondary structure of the ribosomal RNA. These mutations constitute a regulatory step which acclimates the Shine-Dalgarno mechanism to the translational regulation regime of chloroplasts. Interestingly, we show that these chloroplast features occur in modern cyanobacteria as well, promoting the idea that they have evolved prior to endosymbiosis." @default.
• W2896738062 created "2018-10-26" @default.
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• W2896738062 date "2018-10-11" @default.
• W2896738062 modified "2023-09-24" @default.
• W2896738062 title "Solving the riddle of the evolution of Shine-Dalgarno based translation in chloroplasts" @default.
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• W2896738062 doi "https://doi.org/10.1101/440842" @default.
• W2896738062 hasPublicationYear "2018" @default.
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