A small ribosome-associated ncRNA globally inhibits translation by restricting ribosome dynamics.

RNA Biology
Julia ReutherNorbert Polacek

Abstract

Ribosome-associated non-coding RNAs (rancRNAs) have been recognized as an emerging class of regulatory molecules capable of fine-tuning translation in all domains of life. RancRNAs are ideally suited for allowing a swift response to changing environments and are therefore considered pivotal during the first wave of stress adaptation. Previously, we identified an mRNA-derived 18 nucleotides long rancRNA (rancRNA_18) in Saccharomyces cerevisiae that rapidly downregulates protein synthesis during hyperosmotic stress. However, the molecular mechanism of action remained enigmatic. Here, we combine biochemical, genetic, transcriptome-wide and structural evidence, thus revealing rancRNA_18 as global translation inhibitor by targeting the E-site region of the large ribosomal subunit. Ribosomes carrying rancRNA_18 possess decreased affinity for A-site tRNA and impaired structural dynamics. Cumulatively, these discoveries reveal the mode of action of a rancRNA involved in modulating protein biosynthesis at a thus far unequalled precision.

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Methods Mentioned

BETA
FRET
dot blot
RNA-Seq
microscale thermophoresis
RNAseq
density gradient fractionation
transferase
PCR

Software Mentioned

ctffind
featureCounts
AIDA Image Analyse
Motioncor
Chimera
DESeq2
Hisat2
Relion
Vitrobot
ImageQuant TL

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