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Series GSE217180 Query DataSets for GSE217180
Status Public on Apr 12, 2023
Title Translation regulation of specific mRNAs by RPS26 C-terminal RNA-binding tail integrates energy metabolism and AMPK-mTOR signaling
Organism Homo sapiens
Experiment type Expression profiling by high throughput sequencing
Summary Increasing evidence suggests that ribosome composition and modifications contribute to translation control. Much less is known about the regulatory roles of mRNA binding by ribosomal proteins to specific mRNA translation and ribosome specialization. Here we used CRISPR-Cas9, to mutate the RPS26 C-terminal tail (RPS26dC) predicted to bind AUG upstream nucleotides at the exit channel. RPS26 binding to positions -10 to -16 of short 5’UTR mRNAs exerts positive and negative effects on translation directed by Kozak and TISU, respectively. Consistent with that, shortening the 5′UTR from 16 to 10 nt diminished Kozak and enhanced TISU-driven translation. As TISU is resistant and Kozak is sensitive to energy stress, we examined stress responses and found that the RPS26dC mutation confers resistance to glucose starvation and mTOR inhibition. Furthermore, the basal mTOR activity is reduced while AMPK is activated in RPS26dC cells, mirroring energy-deprived WT cells. Likewise, the translatome of RPS26dC cells is correlated to glucose-starved WT cells. Our findings uncover the central roles of RPS26 C-terminal RNA binding in energy metabolism, in the translation of mRNAs bearing specific features and in the translation tolerance of TISU genes to energy stress.
Overall design We used CRISPR-Cas9 mediated mutagenesis in mammalian cells to delete endogenous RPS26 C terminus (RPS26dC cells). For global translation analysis, the collected samples were merged to create the Light (2-5 ribosomes) and Heavy (6+ ribosomes) translated fractions. Equivalent RNA concentrations of Input, Light and Heavy translated ribosome were taken for the library preparation for MARS-Seq.
Contributor(s) Havkin-Solomon T, Reuven N, Shaul Y, Dikstein R
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Submission date Nov 03, 2022
Last update date Apr 12, 2023
Contact name Tal Havkin-Solomon
Phone +972-508311789
Organization name Weizmann Institute of Science
Department Biomolecular Sciences
Lab Rivka Dikstein
Street address 234 Herzl
City Rehovot
ZIP/Postal code 7610001
Country Israel
Platforms (1)
GPL16791 Illumina HiSeq 2500 (Homo sapiens)
Samples (24)
GSM6706293 WT, Glucose starvation, Heavy fraction, replicate 2
GSM6706294 WT, Glucose starvation, Heavy fraction, replicate 1
GSM6706295 WT, Glucose starvation, Input fraction, replicate 2
BioProject PRJNA897861

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Supplementary file Size Download File type/resource
GSE217180_RPS26_project_Feb2021.gene_expression_levels.protein_coding_genes.with_gene_symbol_and_biotype.diff_lanes_sum_togther.RPS26_project.txt.gz 721.4 Kb (ftp)(http) TXT
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