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Depletion of eIF4G from yeast cells narrows the range of translational efficiencies genome-wide

ABSTRACT: Investigation of the effects of genetically depleting eIF4G from yeast cells on global translational efficiencies, using gene expresssion microarrays to measure the abundance of mRNA in polysomes relative to total mRNA for ~5900 genes

ORGANISM(S): Saccharomyces cerevisiae

PROVIDER: GSE25721 | GEO | 2010/12/02

SECONDARY ACCESSION(S): PRJNA133849

REPOSITORIES: GEO

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Depletion of eIF4G from yeast cells narrows the range of translational efficiencies genome-wide

Project description:Investigation of the effects of genetically depleting eIF4G from yeast cells on global translational efficiencies, using gene expresssion microarrays to measure the abundance of mRNA in polysomes relative to total mRNA for ~5900 genes Three biological replicates were examined (designated projects I, II, and III), representing heavy polysomes (HP), light polysomes (LP), and total RNA (T) preparations from three independent pairs of WT and eIF4G degron mutant cultures. Cy3-labeled cDNAs were generated from the 3 HP, 3 LP, and 3 total RNA samples prepared for each strain and the resulting 18 sets of cDNAs were used to probe three (technical) replicate whole-genome microarrays, containing multiple 60-mer oligonucleotides for each gene. The “normalized gene expression summary values” were calculated for each gene from the data obtained from the three technical replicates and used to calculate the translational efficiency (TE) of each gene as the ratio of the intensity values for HP to total RNA (HP/T) or LP to total RNA (LP/T) for each project.

2010-12-02 | E-GEOD-25721 | biostudies-arrayexpress

Transcription profiling analysis of human global and individual mRNA translational efficiencies

Project description:A cell synchronization protocol was established in which global and individual mRNA translational efficiencies could be examined. While global translational efficiency was reduced in mitotic cells, approximately 3% of mRNAs remained predominantly associated with large polysomes during mitosis, as determined by cDNA microarray analyses. The 5 noncoding regions of six mRNAs were shown to contain internal ribosome entry sites (IRES). However, not all known mRNAs that contain IRES elements were actively translated during mitosis, arguing that specific IRES sequences are differentially regulated during mitosis.

2005-10-31 | E-SMDB-2546 | biostudies-arrayexpress

Depletion of eIF4G from yeast cells narrows the range of translational efficiencies genome-wide.

Project description: Not available

| S-EPMC3042410 | biostudies-literature

eIF4B preferentially stimulates translation of long mRNAs with structured 5’UTRs and low closed-loop potential but weak dependence on eIF4G

Project description:DEAD-box RNA helicases eIF4A and Ded1 promote translation by resolving mRNA secondary structures that impede preinitiation complex (PIC) attachment to mRNA or scanning. eIF4B is a cofactor for eIF4A but might also function independently of eIF4A. Ribosome profiling of mutants lacking eIF4B or with impaired eIF4A or Ded1 activity revealed that eliminating eIF4B reduces the relative translational efficiencies of many more genes than does inactivation of eIF4A, despite comparable reductions in bulk translation, and few genes display unusually strong requirements for both factors. However, either eliminating eIF4B or inactivating eIF4A preferentially impacts mRNAs with longer, more structured 5’UTRs. These findings reveal an eIF4A-independent role for eIF4B in addition to its function as eIF4A cofactor in promoting PIC attachment or scanning on structured mRNAs. eIF4B, eIF4A, and Ded1 mutations also preferentially impair translation of longer mRNAs in a fashion mitigated by the ability to form closed-loop mRNPs via eIF4F-Pab1 association, suggesting cooperation between closed-loop assembly and eIF4B/helicase functions. Remarkably, depleting eIF4G, the scaffold subunit of eIF4F, preferentially impacts short mRNAs with strong closed-loop potential and unstructured 5’UTRs, exactly the opposite features associated with hyperdependence on the eIF4B/helicases. We propose that short, highly efficient mRNAs preferentially depend on the stimulatory effects of eIF4G-dependent closed-loop assembly.

2016-05-27 | GSE81966 | GEO

Lifespan extension via eIF4G inhibition is mediated by post-transcriptional remodeling of stress response gene expression in C. elegans

Project description:Reducing protein synthesis slows growth and development but can increase adult lifespan. We demonstrate that knock-down of eukaryotic translation initiation factor 4G (eIF4G), which is down-regulated during starvation, results in differential translation of genes important for growth and longevity in C. elegans. Genome-wide mRNA translation state analysis showed that inhibition of IFG-1, the C. elegans ortholog of eIF4G, results in a relative increase in ribosomal loading and translation of stress response genes. Some of these genes are required for lifespan extension when IFG-1 is inhibited and are new determinants of longevity. Furthermore, enhanced ribosomal loading of certain mRNAs upon IFG-1 inhibition was correlated with increased mRNA length. This association was supported by changes in the proteome assayed via quantitative mass spectrometry. Our results support a role for IFG-1 in mediating the antagonistic effects on growth and somatic maintenance by modulating translation of a specific class of mRNA based on transcript length. 24 experimental samples were analyzed using custom oligo microarrays. A wild type sample pool was used as the Cy3 reference/control for all experimetal samples. All extracted RNA prior to array analysis was fractioned (via a sucrose gradient) based on ribosomal loading and pooled into ribosomal and free RNA (Fraction1), light polysomes (Fraction2) and heavy polysomes (Fraction3) as described in the experimental procedures. The control RNAi is ‘empty’ vector L4440 RNAi feeding vector plasmid (1999 Firelab vector kit) transformed HT115(DE3), which was obtained from the Caenorhabditis Genetics Center (University of Wisconsin).

2011-04-16 | E-GEOD-28665 | biostudies-arrayexpress

Yeast eIF4G homogenic whole cell oxidative stress LC-MSMS

Project description:Translation initiation factor 4G (eIF4G) is an integral component of the eIF4F complex which is key to translation initiation for most eukaryotic mRNAs. Many eIF4G isoforms have been described in diverse eukaryotic organisms but we currently have a poor understanding of their functional roles and whether they regulate translation in an mRNA specific manner. The yeast Saccharomyces cerevisiae expresses two eIF4G isoforms, eIF4G1 and eIF4G2, that have previously been considered as functionally redundant with any phenotypic differences arising due to alteration in eIF4G expression levels. Using homogenic strains that express eIF4G1 or eIF4G2 as the sole eIF4G isoforms at comparable expression levels to total eIF4G, we show that eIF4G1 is specifically required to mediate the translational response to oxidative stress. We use quantitative proteomics to show that eIF4G1 promotes oxidative stress-specific proteome changes.

2023-10-24 | PXD041682 | Pride

Transcription profiling of Saccharomyces cerevisiae eIF4G isoforms to investigate eIF4G deletion transcript abundance

Project description:Effect of eIF4G gene deletion on transcript abundance.

2009-10-24 | E-MEXP-2400 | biostudies-arrayexpress

Transcription profiling of Saccharomyces cerevisiae eIF4G isoforms to investigate eIF4G deletion ribosome occupancy

Project description:Effect of eIF4G gene deletion on the ribosome occupancy.

2009-10-24 | E-MEXP-2399 | biostudies-arrayexpress

Transcription profiling of Saccharomyces cerevisiae eIF4G isoforms to investigate eIF4G homogenic polysome abundance

Project description:Effect of expressing a single eIF4G isoform at wild type levels on relative polysome abundance.

2009-10-14 | E-MEXP-2402 | biostudies-arrayexpress

Systemic remodeling of translation efficiencies on oxygen stimulus.

Project description:Protein concentrations evolve under greater evolutionary constraint than mRNA levels. In addition, translation efficiency of mRNA populations represents the chief determinant of basal protein concentrations. This raises a fundamental question as to how mRNA and protein levels are actively coordinated in dynamic systems responding to acute physiological stimuli. This report examines the contributions of mRNA abundance and translation efficiency to protein output in cells responding to oxygen stimulus. We show that alternative translation efficiencies, not mRNA levels, represent the major adaptive mechanism that governs the cellular response to perturbations in [O2]. This phenomenon is coordinated by eIF4F under atmospheric [O2] and the previously uncharacterized hypoxic eIF4F (eIF4FH) under low [O2]. The oxygen-regulated remodeling of translation efficiency enables oxygenated and hypoxic cells to produce surprisingly divergent translatomes of similar complexity, with minimal dependence on mRNA levels. Changes in mRNA concentrations observed between normoxic and hypoxic cells are likely neutral, as they are during evolution. We propose that mRNAs contain hard-wired translation efficiency determinants for their triage by the translation apparatus on [O2] stimulus.

2016-02-05 | PXD003037 | Pride

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