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SRX3414689: GSM2863256: pat1_t0_rep3; Saccharomyces cerevisiae; OTHER
4 ILLUMINA (NextSeq 500) runs: 20.9M spots, 1.6G bases, 597.1Mb downloads

Submitted by: NCBI (GEO)
Study: Study of ribosome dynamics after osmotic stress in Lsm1 and Pat1 deletions
show Abstracthide Abstract
In this study, we aim to understand how the fates of stress-activated mRNAs are determined under stress conditions by isolating individual osmostress-activated mRNA species, quantitating the proteins associated in vivo with each of them, and analyzing how deletion of these proteins impact on the expression of stress-activated genes. By comparison with the proteome associated with individual not stress-related mRNA species, we show specific proteins to be preferentially binding to osmostress-activated mRNAs, notably members of the cytoplasmic Pat1 / Lsm1 – 7 complex. We evaluated the impact of this complex on the stress response by analyzing expression of osmostress-activated genes, production of osmo-protein and, mapping ribosome transit at single nucleotide resolution using 5'-phosphate sequencing of mRNAs in lsm1 and pat1 mutants. We conclude that our biochemical co-purification approach has successfully identified RNA-binding proteins with a particular role in regulating post-transcriptional expression of stress-activated mRNAs. Overall design: We performed 3 biologically independent 5PSeq experiments before stress (t0) and 30 minutes after osmotic stress (t 30).
Sample: pat1_t0_rep3
SAMN08052641 • SRS2707675 • All experiments • All runs
Library:
Instrument: NextSeq 500
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: SINGLE
Construction protocol: Cells were harvested and pellets were frozen in liquid nitrogen and kept at -80°C until further processing. Total RNA was extracted from yeast cells using phenol:chloroform extraction. 5PSeq methods was performed as previously described (PMID: 26820793) with minor modifications. 6µg of total RNA was used as input. After single-stranded RNA ligation, partial rRNA depletion was performed using Ribo-Zero Magnetic Gold Kit (Epicentre). 5PSeq libraries were PRC amplified (19 cycles), pooled and size selected (300-500nt). Samples were sequenced using a NextSeq500 producing single-end 75 reads.
Experiment attributes:
GEO Accession: GSM2863256
Links:
Runs: 4 runs, 20.9M spots, 1.6G bases, 597.1Mb
Run# of Spots# of BasesSizePublished
SRR63147805,245,001396.8M150.3Mb2018-07-18
SRR63147815,210,352394.2M148.7Mb2018-07-18
SRR63147825,252,862397.4M150.5Mb2018-07-18
SRR63147835,181,230392M147.7Mb2018-07-18

ID:
4758772

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